癞葡萄降血糖肽的制备及其降糖机理研究
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摘要
癞葡萄(Momordica charantia L.Var.abbreviata Ser.,MCV)是葫芦科(Cucurbitaceae)苦瓜属的一个变种,原产于印度东部。由江苏省优质产品开发服务中心驯化改良而成,无锡富仁生物技术有限公司大面积栽培获得成功。现代药理研究证明苦瓜具有降血糖、降血脂、抗癌、抗艾滋病病毒、抑菌、抗氧化等功效。本论文以癞葡萄鲜果为原料,对癞葡萄水提物的降血糖活性进行了研究,并从中筛选出降血糖活性肽MC2-1-5。同时通过碱性蛋白酶Alcalase酶法水解癞葡萄蛋白制备具有降血糖活性的酶解物(MCPHs),并对MC2-1-5和MCPHs的降糖机理进行了研究,研究结果如下:
原料癞葡萄鲜果的化学组成为:水分92.28%,灰分0.48%,粗蛋白1.42%,粗脂肪0.93%,还原糖1.48%,多糖1.55%,粗纤维0.78%。以料水比、盐浓度、提取时间和提取温度为因素,以蛋白提取率为指标,通过正交实验确定癞葡萄水提物的最佳提取工艺条件。综合考虑成本、后续工作量等因素,确定提取条件为料液比1:2,盐浓度0.05 mol/L,提取时间4 h,提取温度40℃。对在此工艺条件下得到的水提物进行氨基酸组成分析,结果表明:水提物中含量最高的氨基酸为精氨酸,其次为谷氨酸和天门冬氨酸,含量分别为22.06%、9.75%、9.54%。三者约占水提物氨基酸总含量的41.35%。新鲜癞葡萄精氨酸、谷氨酸和天门冬氨酸的含量依次为20.13%、12.51%和9.63%,三者约占癞葡萄水提物氨基酸总含量的42.27%。
通过四氧嘧啶型糖尿病小鼠研究了水提物的降血糖作用。结果表明不同剂量(250、500和750 mg/kg)的癞葡萄水提物可显著降低糖尿病小鼠的血糖浓度;水提物的降血糖作用在灌胃后2 h就已开始,并在4 h后达到最大。本实验中的最佳有效剂量为500 mg/kg,该剂量水提物比20 mg/kg剂量的格列本脲片产生的降糖效果更佳。然而,水提物对正常小鼠的血糖浓度不会产生显著影响。此外,水提物还可显著改善糖尿病小鼠的糖耐量。以500 mg/kg剂量连续治疗糖尿病小鼠15 d显著降低小鼠空腹血糖浓度,且和时间成正相关。到第15 d,血糖值较治疗前下降59.88%。水提物还显著增加糖尿病小鼠的体重、降低小鼠的采食量和饮水量。
以小鼠血糖浓度为指标,通过超滤、Sephadex G-25凝胶柱和半制备RP-HPLC从水提物中筛选降血糖肽。研究了进料流速、操作压力、操作温度和料液浓度对膜通量的影响。根据结果选择超滤条件:进料流速200 L/h、温度20℃、压力0.10 MPa、料液浓度20 g/L。超滤将癞葡萄水提物分成浓缩液(MC1)和透过液(MC2)两个组分,动物实验表明MC2对糖尿病小鼠血糖浓度有显著影响,在灌胃2 h和4 h后使小鼠血糖浓度降低46.15%和52.59%。而MC1对血糖浓度影响不显著。
Sephadex G-25凝胶色谱柱将MC2分成六个组分,动物实验表明MC2-1在以20 mg/kg剂量灌胃时即可显著降低四氧嘧啶型糖尿病小鼠的血糖浓度,在灌胃2 h和4 h后使小鼠血糖浓度降低47.43%和51.87%。其他组分对小鼠血糖浓度无显著影响。用半制备RP-HPLC柱将MC2-1分成十个组分,动物实验表明组分MC2-1-1和MC2-1-5在以2 mg/kg剂量灌胃时即可显著降低四氧嘧啶型糖尿病小鼠的血糖浓度。与相应的初始血糖值相比,MC2-1-1在灌胃2 h和4 h后分别使小鼠血糖降低43.84%和54.57%;MC2-1-5在灌胃2 h和4 h后分别使小鼠血糖降低61.70%和69.18%。其他组分对小鼠血糖浓度无显著影响。MC2-1-5还可显著改善小鼠的糖耐量。
红外光谱图显示多肽MC2-1-5在500～4000 cm~(-1)范围具有肽的特征吸收峰。串联飞行时间质谱仪测得MC2-1-5的相对分子质量为3405.5174。蛋白测序仪测得MC2-1-5的N端前10个氨基酸残基排列次序为GHPYYSIKKS。
癞葡萄水溶性蛋白不具备降血糖功效,为充分利用这部分蛋白,酶解癞葡萄蛋白制备降血糖酶解物。以水解度、蛋白回收率和MCPHs的降血糖活性为指标,从六种蛋白酶中选出碱性蛋白酶Alcalase 2.4L水解癞葡萄蛋白。通过单因素试验和响应面分析确定Alcalase水解癞葡萄蛋白的最佳工艺条件:[E]/[S]2.37、pH9.2,温度57℃。在此工艺条件下制备了三种不同DH的MCPHs(8.5%、11%和13.5%),动物实验表明DH为8.5%的MCPHs不具备降血糖活性,DH为11%和13.5%的MCPHs都具有显著的降血糖活性,而DH为11%的MCPHs的降血糖活性高于DH为13.5%的降血糖活性。
根据吸附和解吸效果选择DA201-C大孔吸附树脂对MCPHs进行脱盐。脱盐后MCPHs的含量从脱盐前的76.67%提高至脱盐后的85.69%;MCPHs的回收率为78.38%。脱盐率为92.49%。脱盐后的MCPHs仍有显著的降血糖作用,且降血糖作用较脱盐前稍有提高。
研究了MC2-1和MCPHs的降血糖机理。经MC2-1和MCPHs治疗28 d后,与模型对照组相比,糖尿病小鼠体重显著上升;采食量和饮水量显著下降;脏器/体重比恢复至接近正常水平;空腹血糖值下降;糖耐量显著改善;肝糖原含量显著增加。经MC2-1治疗的糖尿病小鼠的血清胰岛素水平显著提高,但经MCPHs治疗的糖尿病小鼠的血清胰岛素水平未见显著改善。
经MC2-1和MCPHs治疗28 d后,小鼠血清FFA、TC、TG和LDL-C水平下降;HDL-C水平上升,说明MC2-1对糖尿病所引起的高血脂症有一定的治疗作用;小鼠MDA含量下降,SOD活性和GSH-Px活性上升,说明MC2-1和MCPHs能够改善糖尿病小鼠体内的氧化应激机制和提高其抗氧化能力。胰腺、肝脏和肾脏组织形态观察表明MC2-1和MCPHs能改善四氧嘧啶对这些器官细胞造成的损伤。
上述结果表明MC2-1的降糖机理是修复糖尿病小鼠胰岛β细胞、促进胰岛素分泌、增加肝糖原含量及提高小鼠抗氧化能力;MCPHs的降糖机理是修复糖尿病小鼠胰岛β细胞,增加肝糖原含量及提高小鼠抗氧化能力。由于MC2-1-5是MC2-1中的主要降血糖组分,因而推测MC2-1-5的降糖机理与MC2-1类似。
Momordica charantia L.Var.abbreviata Ser.(MCV) is a member of Cucurbitaceae family. Momordica charantia variety was originally growing wild in eastern India.It was domesticated and improved by Jiangsu High-quality Farm Product Development Center,China and has been successfully planted over large areas by Wuxi Furen Biological Science and Technology Co., Ltd.It is reported that Momordica Charantia L.has many pharmacological benefits such as antidiabetic,hypocholesterolemic,hypo-triglyceridemic,antitumor,antibacterial,anti-HIV, anti-inflammatory,antioxidant,etc.In this dissertation,the hypoglycemic effect of an aqueous extract from MCV and a purified peptide called MC2-1-5 with hypoglycemic effect from aqueous extract was stuied.At the same time,MCV protein hydrolysates(MCPHs) with hypoglycemic effect were prepared by Alcalase.The hypoglycemic mechanism of MC2-1-5 and MCPHs was studied in alloxan-induced diabetic mice.
The chemical composition of the fresh MCV fruit were analyzed and found to be as follows:92.28%moisture,0.48%ash,1.42%protein,0.93%fat,1.48%reducing sugar,1.55% of polysaccharide and 0.78%crude fiber.The effect of MCV fruit to water ratio,salt concentration,extraction temperature and extraction time on extraction rate of MCV protein were studied.The parameters affecting protein extraction were optimized by an orthogonal experimental design,that is to say,MCV fruit to water ratio of 1:2,salt concentration of 0.05 mol/L,extraction temperature of 40℃and extraction time of 4 h.Amino acid(AA) composition profile of MCV aqueous extract showed that the content of Arginine was highest, followed by Glutamic acid and Aspartic acid of 22.06%,9.75%and 9.54%respectively.The total percentage composition of the three AA was 41.35%.The AA composition profile of fresh MCV showed that the content of Arginine,Glutamic acid and Aspartic acid was 20.13%, 12.51%and 9.63%respectively.The total percentage composition of the three AA was 42.27%.
The acute(a single dose) and chronic(15 d) hypoglycemic effects of aqueous extract were studied in alloxan-induced diabetic mice.The results showed that an aqueous extract from MCV fruit at different doses(250,500 and 750 mg/kg) produced a significant hypoglycemic effect in alloxan-induced diabetic mice within 2 h after oral administration,which was maintained for 4 h.As far as the most effective dose under the experimental conditions stuied is concerned,it was found to be 500 mg/kg in all groups.The hypoglycemic effect of the aqueous extract administered at a dose of 500 mg/kg was more effective than that of glibenclamide administered at a dose of 100 mg/kg.However,the aqueous extract didn't have significant effect on blood glucose level(BGL) in normal mice.In addition,the hypoglycemic effect of aqueous extract was further confirmed by the oral glucose tolerance test(OGTT).A 15-day treatment of the aqueous extract at a dose of 500 mg/kg in diabetic mice continuously lowered the fasting blood glucose(FBG) level.Treatment of alloxan-induced mice for 15 days with the aqueous extract at a dose of 500 mg/kg produced a 59.88%fall in FBG level.During the treatment period,the aqueous extract significantly increased the body weight(BW),lowered the food and water intake in diabetic mice.
Using BGL in diabetic mice as an index,the peptide with hypoglycemic effect was purified by ultrafiltration(UF),Sephadex G-25 and semi-RP-HPLC.The influence of input flow rate,operating pressure,operating temperature and sample concentration on flux was studied.The UF conditions used were:input flow rate 200 L/h,temperature 20℃,pressure 0.1 MPa and sample concentration 20 g/L.On separating MCV aqueous extract into two fractions (MC1 and MC2) by UF,MC2 showed significant hypoglycemic effect in alloxan-induced mice. It produced a reduction of 46.15%and 52.59%in BGL after 2 and 4 h respectively of oral administration.MC1 didn't show hypoglycemic effect in diabetic mice.
Six different fractions of MC2 were obtained by Sephadex G-25 gel filtration chromatography.The in vivo test showed MC 2-1 at a dose of 20 mg/kg could lower the BGL in diabetic mice.It produced a reduction of 47.43%and 51.87%in BGL after 2 and 4 h respectively of oral administration.Other fractions had no significant effect on BGL in diabetic mice.A further ten different fractions of MC 2-1 were obtained through Semi-HPLC.The in vivo test showed MC 2-1-1 and MC 2-1-5 at a dose of 2 mg/kg could lower the BGL in diabetic mice,while other fractions had no significant effect on BGL in diabetic mice.MC2-1-1 produced a reduction of 43.84%and 54.57%in BGL after 2 and 4 h respectively of oral administration.MC2-1-5 produced a reduction of 61.70%and 69.18%in BGL after 2 and 4 h respectively of oral administration.In addition,MC2-1-5 improved the oral glucose tolerance in diabetic mice.
An Infrared spectrograph showed MC2-1-5 to have characteristic IR absorption peak of peptide in the spectral region 500～4000 cm~(-1).MALDI TOF-MS showed the molecular weight of MC2-1-5 was 3405.5174 Da.Protein Sequencer showed the N terminal frist ten AAs of MC2-1-5 were GHPYYSIKKS.
MCV souble protein did not have hypoglycemic effect.In order to make use of the MCV soluble protein,the protein was hydrolyzed to produce MCPHs with hypoglycemic effect.The MCV protein was hydrolyzed using six different proteases.The results showed Alcalase 2.4L to have the best hydrolysing capacity.The protein recovery was also highest with Alcalase.The hydrolysis parameters using Alcalase were optimized as follows:enzyme to substrate ratio 2.37%,pH 9.2 and hydrolysis temperature 57℃.Three hydrolysates with different DH(8.5%, 11%and 13.5%) were obtained using Alcalase under the optimized conditions.The hypoglycemic effect of the three hydrolysates was stuied in alloxan-induced diabetic mice.The results showed that the MCPHs at DH 11%had the best hypoglycemic effect,followed by MCPHs at DH 13.5%.MCPHs at DH 8.5%didn't have any hypoglycemic effect.
DA201-C Macroporous resin was chosen to desalt MCPHs by comparing the adsorbing and desorbing capacity.The optimal conditions for desalting using DA201-C macroporous resin were determined.Under the optimal conditions,the desalting rate and recovery of MCPHs was 92.49%and 82.38%respectively.The content of MCPHs increased from 76.67%to 85.69%.The hypoglycemic effect of desalted MCPHs was better than that of MCPHs before desalting.
The hypoglycemic mechanism of MC2-1 and MCPHs was studied.Compared with the diabetic control,a 28-d treatment with MC2-1 and MCPHs increased the BW and hepatic glycogen content,lowered food intake and water intake,made the organ/body weight near to normal and improved the oral glucose test in diabetic mice.In addition,MC2-1 in particular, increased the serum insulin level in diabetic mice.
MC2-1 and MCPHs could improve the lipid profile in diabetic mice.After a 28-d treatment with MC2-1 and MCPHs,free amino acid(FFA),total cholesterol(TC),triglyceride (TG) and low density lipoprotein cholesterol(LDL-C) lowered significantly,while high density lipoprotein cholesterol(HDL-C) increased significantly.MC2-1 and MCPHs could improve the mechanisms of oxidative stress and antioxidant capacity in diabetic mice.After a 28-d treatment with MC2-1 and MCPHs,malondialdehyde(MDA) content lowered while SOD activity and GSH-Px activity increased.Tissue morphology showed that MC2-1 and MCPHs could improve the cell injury in islet,liver and kidney induced by alloxan in diabetic mice.
In summary,the hypoglycemic mechanism of MC2-1 is to recover the isletβcell,so as to stimulate insulin secretion,to increase the hepatic glycogen content and improve antioxidant capacity in diabetic mice.The hypoglycemic mechanism of MCPHs is similar to that of MC2-1, except that it couldn't stimulate insulin secretion.Since MC2-1-5 is the main hypoglycemic constituent in MC2-1,it is speculated that the hypoglycemic effect of MC2-1-5 was similar to that of MC2-1.
原料癞葡萄鲜果的化学组成为:水分92.28%,灰分0.48%,粗蛋白1.42%,粗脂肪0.93%,还原糖1.48%,多糖1.55%,粗纤维0.78%。以料水比、盐浓度、提取时间和提取温度为因素,以蛋白提取率为指标,通过正交实验确定癞葡萄水提物的最佳提取工艺条件。综合考虑成本、后续工作量等因素,确定提取条件为料液比1:2,盐浓度0.05 mol/L,提取时间4 h,提取温度40℃。对在此工艺条件下得到的水提物进行氨基酸组成分析,结果表明:水提物中含量最高的氨基酸为精氨酸,其次为谷氨酸和天门冬氨酸,含量分别为22.06%、9.75%、9.54%。三者约占水提物氨基酸总含量的41.35%。新鲜癞葡萄精氨酸、谷氨酸和天门冬氨酸的含量依次为20.13%、12.51%和9.63%,三者约占癞葡萄水提物氨基酸总含量的42.27%。
通过四氧嘧啶型糖尿病小鼠研究了水提物的降血糖作用。结果表明不同剂量(250、500和750 mg/kg)的癞葡萄水提物可显著降低糖尿病小鼠的血糖浓度;水提物的降血糖作用在灌胃后2 h就已开始,并在4 h后达到最大。本实验中的最佳有效剂量为500 mg/kg,该剂量水提物比20 mg/kg剂量的格列本脲片产生的降糖效果更佳。然而,水提物对正常小鼠的血糖浓度不会产生显著影响。此外,水提物还可显著改善糖尿病小鼠的糖耐量。以500 mg/kg剂量连续治疗糖尿病小鼠15 d显著降低小鼠空腹血糖浓度,且和时间成正相关。到第15 d,血糖值较治疗前下降59.88%。水提物还显著增加糖尿病小鼠的体重、降低小鼠的采食量和饮水量。
以小鼠血糖浓度为指标,通过超滤、Sephadex G-25凝胶柱和半制备RP-HPLC从水提物中筛选降血糖肽。研究了进料流速、操作压力、操作温度和料液浓度对膜通量的影响。根据结果选择超滤条件:进料流速200 L/h、温度20℃、压力0.10 MPa、料液浓度20 g/L。超滤将癞葡萄水提物分成浓缩液(MC1)和透过液(MC2)两个组分,动物实验表明MC2对糖尿病小鼠血糖浓度有显著影响,在灌胃2 h和4 h后使小鼠血糖浓度降低46.15%和52.59%。而MC1对血糖浓度影响不显著。
Sephadex G-25凝胶色谱柱将MC2分成六个组分,动物实验表明MC2-1在以20 mg/kg剂量灌胃时即可显著降低四氧嘧啶型糖尿病小鼠的血糖浓度,在灌胃2 h和4 h后使小鼠血糖浓度降低47.43%和51.87%。其他组分对小鼠血糖浓度无显著影响。用半制备RP-HPLC柱将MC2-1分成十个组分,动物实验表明组分MC2-1-1和MC2-1-5在以2 mg/kg剂量灌胃时即可显著降低四氧嘧啶型糖尿病小鼠的血糖浓度。与相应的初始血糖值相比,MC2-1-1在灌胃2 h和4 h后分别使小鼠血糖降低43.84%和54.57%;MC2-1-5在灌胃2 h和4 h后分别使小鼠血糖降低61.70%和69.18%。其他组分对小鼠血糖浓度无显著影响。MC2-1-5还可显著改善小鼠的糖耐量。
红外光谱图显示多肽MC2-1-5在500～4000 cm~(-1)范围具有肽的特征吸收峰。串联飞行时间质谱仪测得MC2-1-5的相对分子质量为3405.5174。蛋白测序仪测得MC2-1-5的N端前10个氨基酸残基排列次序为GHPYYSIKKS。
癞葡萄水溶性蛋白不具备降血糖功效,为充分利用这部分蛋白,酶解癞葡萄蛋白制备降血糖酶解物。以水解度、蛋白回收率和MCPHs的降血糖活性为指标,从六种蛋白酶中选出碱性蛋白酶Alcalase 2.4L水解癞葡萄蛋白。通过单因素试验和响应面分析确定Alcalase水解癞葡萄蛋白的最佳工艺条件:[E]/[S]2.37、pH9.2,温度57℃。在此工艺条件下制备了三种不同DH的MCPHs(8.5%、11%和13.5%),动物实验表明DH为8.5%的MCPHs不具备降血糖活性,DH为11%和13.5%的MCPHs都具有显著的降血糖活性,而DH为11%的MCPHs的降血糖活性高于DH为13.5%的降血糖活性。
根据吸附和解吸效果选择DA201-C大孔吸附树脂对MCPHs进行脱盐。脱盐后MCPHs的含量从脱盐前的76.67%提高至脱盐后的85.69%;MCPHs的回收率为78.38%。脱盐率为92.49%。脱盐后的MCPHs仍有显著的降血糖作用,且降血糖作用较脱盐前稍有提高。
研究了MC2-1和MCPHs的降血糖机理。经MC2-1和MCPHs治疗28 d后,与模型对照组相比,糖尿病小鼠体重显著上升;采食量和饮水量显著下降;脏器/体重比恢复至接近正常水平;空腹血糖值下降;糖耐量显著改善;肝糖原含量显著增加。经MC2-1治疗的糖尿病小鼠的血清胰岛素水平显著提高,但经MCPHs治疗的糖尿病小鼠的血清胰岛素水平未见显著改善。
经MC2-1和MCPHs治疗28 d后,小鼠血清FFA、TC、TG和LDL-C水平下降;HDL-C水平上升,说明MC2-1对糖尿病所引起的高血脂症有一定的治疗作用;小鼠MDA含量下降,SOD活性和GSH-Px活性上升,说明MC2-1和MCPHs能够改善糖尿病小鼠体内的氧化应激机制和提高其抗氧化能力。胰腺、肝脏和肾脏组织形态观察表明MC2-1和MCPHs能改善四氧嘧啶对这些器官细胞造成的损伤。
上述结果表明MC2-1的降糖机理是修复糖尿病小鼠胰岛β细胞、促进胰岛素分泌、增加肝糖原含量及提高小鼠抗氧化能力;MCPHs的降糖机理是修复糖尿病小鼠胰岛β细胞,增加肝糖原含量及提高小鼠抗氧化能力。由于MC2-1-5是MC2-1中的主要降血糖组分,因而推测MC2-1-5的降糖机理与MC2-1类似。
Momordica charantia L.Var.abbreviata Ser.(MCV) is a member of Cucurbitaceae family. Momordica charantia variety was originally growing wild in eastern India.It was domesticated and improved by Jiangsu High-quality Farm Product Development Center,China and has been successfully planted over large areas by Wuxi Furen Biological Science and Technology Co., Ltd.It is reported that Momordica Charantia L.has many pharmacological benefits such as antidiabetic,hypocholesterolemic,hypo-triglyceridemic,antitumor,antibacterial,anti-HIV, anti-inflammatory,antioxidant,etc.In this dissertation,the hypoglycemic effect of an aqueous extract from MCV and a purified peptide called MC2-1-5 with hypoglycemic effect from aqueous extract was stuied.At the same time,MCV protein hydrolysates(MCPHs) with hypoglycemic effect were prepared by Alcalase.The hypoglycemic mechanism of MC2-1-5 and MCPHs was studied in alloxan-induced diabetic mice.
The chemical composition of the fresh MCV fruit were analyzed and found to be as follows:92.28%moisture,0.48%ash,1.42%protein,0.93%fat,1.48%reducing sugar,1.55% of polysaccharide and 0.78%crude fiber.The effect of MCV fruit to water ratio,salt concentration,extraction temperature and extraction time on extraction rate of MCV protein were studied.The parameters affecting protein extraction were optimized by an orthogonal experimental design,that is to say,MCV fruit to water ratio of 1:2,salt concentration of 0.05 mol/L,extraction temperature of 40℃and extraction time of 4 h.Amino acid(AA) composition profile of MCV aqueous extract showed that the content of Arginine was highest, followed by Glutamic acid and Aspartic acid of 22.06%,9.75%and 9.54%respectively.The total percentage composition of the three AA was 41.35%.The AA composition profile of fresh MCV showed that the content of Arginine,Glutamic acid and Aspartic acid was 20.13%, 12.51%and 9.63%respectively.The total percentage composition of the three AA was 42.27%.
The acute(a single dose) and chronic(15 d) hypoglycemic effects of aqueous extract were studied in alloxan-induced diabetic mice.The results showed that an aqueous extract from MCV fruit at different doses(250,500 and 750 mg/kg) produced a significant hypoglycemic effect in alloxan-induced diabetic mice within 2 h after oral administration,which was maintained for 4 h.As far as the most effective dose under the experimental conditions stuied is concerned,it was found to be 500 mg/kg in all groups.The hypoglycemic effect of the aqueous extract administered at a dose of 500 mg/kg was more effective than that of glibenclamide administered at a dose of 100 mg/kg.However,the aqueous extract didn't have significant effect on blood glucose level(BGL) in normal mice.In addition,the hypoglycemic effect of aqueous extract was further confirmed by the oral glucose tolerance test(OGTT).A 15-day treatment of the aqueous extract at a dose of 500 mg/kg in diabetic mice continuously lowered the fasting blood glucose(FBG) level.Treatment of alloxan-induced mice for 15 days with the aqueous extract at a dose of 500 mg/kg produced a 59.88%fall in FBG level.During the treatment period,the aqueous extract significantly increased the body weight(BW),lowered the food and water intake in diabetic mice.
Using BGL in diabetic mice as an index,the peptide with hypoglycemic effect was purified by ultrafiltration(UF),Sephadex G-25 and semi-RP-HPLC.The influence of input flow rate,operating pressure,operating temperature and sample concentration on flux was studied.The UF conditions used were:input flow rate 200 L/h,temperature 20℃,pressure 0.1 MPa and sample concentration 20 g/L.On separating MCV aqueous extract into two fractions (MC1 and MC2) by UF,MC2 showed significant hypoglycemic effect in alloxan-induced mice. It produced a reduction of 46.15%and 52.59%in BGL after 2 and 4 h respectively of oral administration.MC1 didn't show hypoglycemic effect in diabetic mice.
Six different fractions of MC2 were obtained by Sephadex G-25 gel filtration chromatography.The in vivo test showed MC 2-1 at a dose of 20 mg/kg could lower the BGL in diabetic mice.It produced a reduction of 47.43%and 51.87%in BGL after 2 and 4 h respectively of oral administration.Other fractions had no significant effect on BGL in diabetic mice.A further ten different fractions of MC 2-1 were obtained through Semi-HPLC.The in vivo test showed MC 2-1-1 and MC 2-1-5 at a dose of 2 mg/kg could lower the BGL in diabetic mice,while other fractions had no significant effect on BGL in diabetic mice.MC2-1-1 produced a reduction of 43.84%and 54.57%in BGL after 2 and 4 h respectively of oral administration.MC2-1-5 produced a reduction of 61.70%and 69.18%in BGL after 2 and 4 h respectively of oral administration.In addition,MC2-1-5 improved the oral glucose tolerance in diabetic mice.
An Infrared spectrograph showed MC2-1-5 to have characteristic IR absorption peak of peptide in the spectral region 500～4000 cm~(-1).MALDI TOF-MS showed the molecular weight of MC2-1-5 was 3405.5174 Da.Protein Sequencer showed the N terminal frist ten AAs of MC2-1-5 were GHPYYSIKKS.
MCV souble protein did not have hypoglycemic effect.In order to make use of the MCV soluble protein,the protein was hydrolyzed to produce MCPHs with hypoglycemic effect.The MCV protein was hydrolyzed using six different proteases.The results showed Alcalase 2.4L to have the best hydrolysing capacity.The protein recovery was also highest with Alcalase.The hydrolysis parameters using Alcalase were optimized as follows:enzyme to substrate ratio 2.37%,pH 9.2 and hydrolysis temperature 57℃.Three hydrolysates with different DH(8.5%, 11%and 13.5%) were obtained using Alcalase under the optimized conditions.The hypoglycemic effect of the three hydrolysates was stuied in alloxan-induced diabetic mice.The results showed that the MCPHs at DH 11%had the best hypoglycemic effect,followed by MCPHs at DH 13.5%.MCPHs at DH 8.5%didn't have any hypoglycemic effect.
DA201-C Macroporous resin was chosen to desalt MCPHs by comparing the adsorbing and desorbing capacity.The optimal conditions for desalting using DA201-C macroporous resin were determined.Under the optimal conditions,the desalting rate and recovery of MCPHs was 92.49%and 82.38%respectively.The content of MCPHs increased from 76.67%to 85.69%.The hypoglycemic effect of desalted MCPHs was better than that of MCPHs before desalting.
The hypoglycemic mechanism of MC2-1 and MCPHs was studied.Compared with the diabetic control,a 28-d treatment with MC2-1 and MCPHs increased the BW and hepatic glycogen content,lowered food intake and water intake,made the organ/body weight near to normal and improved the oral glucose test in diabetic mice.In addition,MC2-1 in particular, increased the serum insulin level in diabetic mice.
MC2-1 and MCPHs could improve the lipid profile in diabetic mice.After a 28-d treatment with MC2-1 and MCPHs,free amino acid(FFA),total cholesterol(TC),triglyceride (TG) and low density lipoprotein cholesterol(LDL-C) lowered significantly,while high density lipoprotein cholesterol(HDL-C) increased significantly.MC2-1 and MCPHs could improve the mechanisms of oxidative stress and antioxidant capacity in diabetic mice.After a 28-d treatment with MC2-1 and MCPHs,malondialdehyde(MDA) content lowered while SOD activity and GSH-Px activity increased.Tissue morphology showed that MC2-1 and MCPHs could improve the cell injury in islet,liver and kidney induced by alloxan in diabetic mice.
In summary,the hypoglycemic mechanism of MC2-1 is to recover the isletβcell,so as to stimulate insulin secretion,to increase the hepatic glycogen content and improve antioxidant capacity in diabetic mice.The hypoglycemic mechanism of MCPHs is similar to that of MC2-1, except that it couldn't stimulate insulin secretion.Since MC2-1-5 is the main hypoglycemic constituent in MC2-1,it is speculated that the hypoglycemic effect of MC2-1-5 was similar to that of MC2-1.
引文
1 江苏省植物研究所.江苏植物志(下册)[M].南京:江苏科学技术出版社,1982,810
2 江苏新医学院.中药大辞典[M].上海:上海科学技术出版社,1980,1289-1290
3 程霜,崔庆新,王勇.水溶性苦瓜多糖的提取与鉴定[J].郑州粮食学院学报,2000,21(2):53-56
4(明)李时珍.本草纲目[M].北京:人民卫生出版社,1982
5 田亚维.苦瓜的研究概述[J].医学信息,2002,15(10):619-670
6 邓向军,徐斌,董英.苦瓜化学成分的研究进展[J].时珍国医国药,2006,17(12):2449-2451
7 Okabe H,Miyahara Y,Yamauchi T,et al.Studies on the constituent of Momordica charantia L.isolation and characterization of Momordicosides A and B[J].Chem Pharm Bull,1980,28:2753-2762
8 Miyahara Y,Okabe H,Yamauchi T.Studies on the Constituent of Momordica charantia L.Ⅱ.Isolation and Characterization of minor seed glycosides,Momordicosides C,D and E[J].Chem Pharm Bull,1981,29(6):1561-1566
9 Yasuda M,Iwamoto M,Okabe H,et al.Structures of momordicines Ⅰ,Ⅱand Ⅲ:the bitter principles in the leaves and vines of Momordica charantia L[J].Chem Pharm Bul,1984,32(5):2044-2047
10 Murakami T,Emoto A,Matsuda H,et al.Structures of New Cucurbitane-Type Triterpene Glycosides,Goyaglycosides-a,-b,-c,-d,-e,-f,-g,and -h,and New Oleanane-Type Triterpene Saponins,Goyasaponins Ⅰ,Ⅱ,and Ⅲ,from the fresh fruit of Japanese Momordica charantia L.[J].Pharmaceutical Society of Japan,2001,49(1):54-63
11 Raman A,Lau C.Anti-diabetic properties and phytochemistry of Momordica charantia L.(Cucurbitaceae)[J].Phytomedicine,1996,2(4):349-362
12 Guevara AP,Linn Sylianco C,Dayrit F,et al.Antimutagens from Momordica charantia[J]Mutat Res,1990,230(2):120-123
13 Sucrow W.Uber steringlucoside und ein neues stigmastadienol aus Momordica charantia[J].Tetrahedron Lette,1965,26(9):2217
14 Lotlikar MM,Rajarama RMR.Pharmacology of a hypoglycemic principle:isolated from the fruits of Momordica charantia Linn[J].Indian Journal of Pharmacy,1966,28:129-133
15 Khanna P,Jain C,Bansal R.Effect of cholesterol on growth and production of diosgenin,gitogenin,titogenin and sterols in suspension cultures[J].Indian J Exp Biol,1975,13:211-213
16 Lau C,Raman A,Noel M,et al.Phytochemicals isolated from the anti-hyperglycaemic hexane extract of the unripe fruit of Momordica charantia L[J].J Pharm Pharmacol Suppl,1998,50:84
17 常凤岗.苦瓜的化学成分研究[J].中草药,1995,26(10):507-509
18 Khanna P,Jain SC.Panangariya A.Hyopglycemic activity of polypeotide-P from a plant source[J].Nat Prod,1981,44:648-655
19 Mokkhasmit M,Swatdimongkol K,Satrawaha P.Study on toxicity of Thai medicinal plants[J].Bull Dept Med Sci,1971,12(2/4):36-65
20 Nag B,Medicherla S,Sharma,SD.Orally active fraction of Momordica charantia,active peptides thereof,and their use in the treatment of diabetes[P].U.S.(P) US6127338,2000-10-03
21 Tomita M,Kurokawa T,Onozaki K,et al.Purification of galactose-binding phytoagglutinins and phytotoxin by affinity column chromatography using sepharose[J].Experientia,1972,28(1):84-85
22 Montecucchi PC,Lazzarin AM,Barbieri L,et al.N-termial sequence of some ribosome inactivating protein[J].Int J Pept Protein Res,1989,33:263-267
23 Kubota S,Yeung HW,Yang JT.Conformation of abortifacient proteins:trichosanthin,α-momorcharin and β-momorcharin[J].Biochemica et biophysica acta,1986,871:101-106
24 Pu Z,Lu BY,Liu WY,et al.Characterization of the enzymatic mechanism of gamma-momorcharin,a novel ribosome-inactivating protein with lower mocular weight of 11,5000 purified from the seeds of bitter gourd[J].Biochem Biophys Res Commun,1996,229(1):287-294
25 Barbieri L.Inhibition of protein synthesis in vitro by proteins from the seeds of Momoridca charantia[J].Biocetem J,1980,186:443-452
26 Lee-Huang S,Huang PL,Nara PL.MAP 30:a new inhibitor of HIV-1 infection and replication[J].FEBS lett,1990,272:12-18
27 Jiratchariyakul W,Wiwat C,Vongsakul M,et al.HIV inhibitor from Thai bitter gourd[J].Planta Med,2001,67(4):350-353
28 彭爱芝.苦瓜中氨基酸和无机元素的含量分析[J].湖南医科大学学报,1996,21(4):305
29 于长春,郑长春,王庆华等.苦瓜籽中脂肪酸的分离及生理作用研究[J].松辽学刊(自然科学版),1995,2:40-41
30 El-Gengaihi S,Karawya MS,Selim MA,et al.A novel pyrimidine glycoside from Momordica charantia L.[J].Pharmazie,1995,50(5):361-362
31 郭亦然,王水,唐琳等.苦瓜籽的生药学研究[J].四川大学学报(自然科学版),2001,38(3):454-456
32 李梦华,商学义.苦瓜对大白鼠血糖的影响[J].天津师大学报(自然科学版),1992,20(2):52-54
33 Akhtar S,Akhtar,AM,Yaqub M.Effect of Momordica charantia on blood glucose level of normal and alloxan-diabetic rabbits[J].Planta Medica,1981,42(3):205-212
34 Hang YY,Zhou YF,Chen ZM,et al.Pharmacological experiments of freeze-dried powder of balsam pear (Momordica charantia L.) on lowering the blood glucose level to mice and its acute toxicity[J].J Plant Resour & Environ,2000,9:19-21
35 黄小琴,杨延莉.苦瓜胶囊降血糖作用研究[J].药学服务与研究,2002,2(2):84-85
36 Reyes BA,Bautista ND,Tanquilut NC,et al.Anti-diabetic potentials of Momordica charantia and Andrographis paniculata and their effects on estrous cyclicity of alloxan-induced diabetic rats[J].J Ethnopharmacol,2006,105(1-2):196-200
37 Karunanayake EH,Jeevathayaparan S,Tennekoon KH,et al.Effect of Momordica charantia fruit juice on the streptozotocin-induced diabetic rats[J].J Ethnopharmacol,1990,30(2):199-204
38 Ahmed I,Adeghate E,Sharma AK,et al.Effects of Momordica charantia fruit juice on islet morphology in the pancreas of the streptozotocin-diabetic rat[J].Diabetes Research Clinical Practices,1998,40:145-151
39 Ahmed I,Lakhani MS,Gillett M,et al.Hypotriglyceridemic and hypocholesterolemic effects of anti-diabetic Momordica charantia(karela) fruit extract in streptozotocin-induced diabetic rats[J].Diabetes Research Clinical Practices,2001,51:155-161
40 Shetty AK,Kumar GS,Sambaiah K,et al.Effect of bitter gourd(Momordica charantia) on glycaemic status in streptozotocin induced diabetic rats[J].Plant Foods Hum Nutr,2005,60(3):109-112
41 Ojewole JA,Adewole SO,Olayiwola G,et al.Hypoglycaemic and hypotensive effects of Momordica charantia Linn(Cucurbitaceae) whole-plant aqueous extract in rats[J].Cardiovasc J S Afr,2006,17(5):227-32
42 Liaquat Ali,Abul Kalam Azad Khan,Muhammad lqbal Rouf Mamun,et al.Studies on hypoglycemic effects of fruit pulp,seed and whole plant of Momordica charantia on normal and diabetic model rats[J].Planta Med,1993,59:408-412
43 Kumar Shetty A,Suresh Kumar G,Veerayya Salimath P,et al.Bitter gourd(Momordica charantia)modulates activities of intestinal and renal disaccharidases in streptozotocin-induced diabetic rats[J].Mol Nutr Food Res,2005,49(8):791-796
44 Jayasooriya AP,Sakono M,Yukizaki C,et al.Effects of Momordica charantia power on serum glucose levels and various lipid parameters in rats fed with cholesterol-free and cholesterol-enriched diets[J].J Ethnopharmacol,2000,72:331-336
45 Miura T,Itoh C,Iwamoto N,et al.Hypoglycemic activity of the fruit of the Momordica charantia in type 2 diabetic mice[J].J Nutr Sci Vitaminol,2001,47(5):340-344
46 Chen QX,Chan LLY,Li-ETS,et al.Bitter melon(Momordica charantia) reduces adiposity,lowers serum insulin and normalizes glucose tolerance in rats fed a high fat diet[J].J Nutr,2003,133:1088-1093
47 Welihinda J,Karunanayake EH,Sheriff MH,et al.Effect of Momordica charantia on the glucose tolerance in maturity onset diabetes[J].J Ethnopharmacol,1986,17:277-282
48 Day C,Cartwright T,Provost J,et al.Hypoglycaemic effect of Momordica charantia extracts[J].Planta Medica,56:426-429
49 Cakici I,Hurmoglu C,Tunctan B,et al.Hypoglycaemic effect of Momordica charantia extracts in normoglycaemic or cyproheptadineinduced hyperglycaemic mice[J].J Ethnopharmacol,1994,44:117-121
50 Raza H,Ahmed I,John A,et al.Modulation of xenobiotic metabolism and oxidative stress in chronic streptozotocin-induced diabetic rats fed with Momordica charantia fruit extract[J].J Biochem Mol Toxicol,2000,14(3):131-139
51 范玉玲,崔福德.苦瓜有效部位降糖活性的比较研究[J].沈阳药科大学学报,2001,18(1):50-52
52 Vikrant V,Grover JK,Tandon N,et al.Treatment with extracts of Momordica charantia and Eugenia jambolana prevents hyperglycemia and hyperinsulinemia in fructose fed rats[J].J Ethnopharmacol,2001,76(2):139-143
53 Kar A,Choudhary BK,Bandyopadhyay NG.Comparative evaluation of hypoglycaemic activity of some Indian medicinal plants in alloxan diabetic rats[J].J Ethnopharmacol,2003,84:105-108
54 Khan B,Arayne MS,Naz S,et al.Hypogylcemic activity of aqueous extract of some indigenous plants[J].Pak J Pharm Sci,2005,18(1):62-64
55 Mahomoodally MF,Gurib-Fakim A,Subratty AH.Effect of exogenous ATP on Momordica charantia Linn.(Cucurbitaceae) induced inhibition of d-glucose,1-tyrosine and fluid transport across rat everted intestinal sacs in vitro[J].J Ethnopharmacol,2007,110(2):257-263
56 Abd El Sattar El Batran S,El-Gengaihi SE,El Shabrawy OA.Some toxicological studies of Momordica charantia L.on albino rats in normal and alloxan diabetic rats[J].J Ethnopharmacol,2006,108(2):236-242
57 Sarkar S,Pranava M,Marita R.Demonstration of the hypoglycemic action of Momordica charantia in validated animal model of diabetes[J].Pharmacol Res,1996,33(1):1-4
58 王先远,金宏.苦瓜皂甙降血糖作用及其机制初探[J].氨基酸和生物资源,2001,23(3):42-45
59 Higashino H,Suzuki A,Tanaka Y,et al.Hypoglycemic effects of Siamese Momordica charantia and Phyllanthus urinaria extracts in streptozotocin induced diabetic rats[J].Nippon Yakurigaku Zasshi,1992,100:415-421
60 Chaturvedi P,George S,Milinganyo M,et al.Effect of Momordica charantia on lipid profile and oral glucose tolerance in diabetic rats[J].Phytother Res,2004,18(11):954-956
61 Tongia A,Tongia S,Dave M,et al.Phytochemical determination and extraction of Momordica charantia fruit and its hypoglycemic potentiation of oral hypoglycemic drugs in diabetes mellitus(NIDDM)[J].Indian J Physiol Pharmacol,2004,48:241-244
62 Lotlikar MM,Rajarama Rao MR.Pharmacology of a hypoglycemic principle isolated from the fruits of Momordica charantia L[J].Indian J Pharm,1966,(28):129-132
63 Olaniyi AA.A neutral constituent of Momordica foetida[J].Lloydia,1975,38:361-362
64 Ng TB,Wong CM,Li WW,et al.A steryl glycoside fractions from Momordica charantia seeds with an inhibitory action on lipid metabolism in vitro[J].Biochem Cell Biol,1986,64(8):766-771
65 张萍萍,王峰,薛爱琴等.苦瓜素降血糖作用的试验研究[J].江苏中医,1997,7:30-31
66 孙宝莹,孙宝全.苦瓜皂甙的药效学实验研究[J].河南中医药学刊.1994,9(6):19-21
67 Matsuda H,Li Y,Murakami T,et al.Antidiabetic principles of natural medicines Ⅲ.Structure-related inhibitory activity and action model of oleanolic acid glycosides on hypoglycemic activity[J].Chem Pharm Bull,1998,46(9):1399-1403
68 赵海雯.苦瓜皂甙的分离纯化及降血糖功效研究[D].硕士学位论文,江南大学,2005
69 石雪萍.苦瓜皂甙的分离、纯化、结构鉴定及降血糖功能研究[D].博士学位论文,江南大学,2007
70 Baldwa VS,Goyal RK,Bhandari CM,et al.A clinical trial of insulin obtained from vegetable source (plant insulin) in patients with diabetes mellitus[J].Rajasthan Med J,1976,15(1):54-60
71 张世荣,姚文贞,薛庆富等.苦瓜降糖肽Ⅰ.分离纯化和特性[J].生物化学与生物物理学报,1980,12(4):291
72 王敬文,龚桂,陈随军等.植物降血糖活性蛋白新组分PA及其制备.专利申请号:93119351,1995-5
73 权建新,篙兴厚,张振强等.苦瓜果实中植物胰岛素降血糖研究.陕西医学杂志,1991,20(11):691-693
74 盛清凯,姚惠源.苦瓜中植物胰岛素的分离及其氨基酸序列测定[J].食品与发酵工业,2004,30(7):129-131
75 王勇庆.苦瓜降血糖作用研究[J].湖南中医杂志,1998,14:54-55
76 Zheng ZX,Teng JY,Liu JY,et al.The hypoglycemic effects of crude polysaccharides extract from Momordica charantia in mice[J].Wei Sheng Yan Jiu,2005,34(3):361-363
77 American Diabetes Association.Screening for type 2 diabetes.Diabetic Care,1998,21:S20-S31
78 World Health Organization,1994.Prevention of diabetes mellitus.Technical Report Series NO 844.Geneva,WHO
79 Grover JK,Rathi SS,Vats V.Amelioration of experimental diabetic neuropathy and gastropathy in rats following oral administration of plant(Eugenia Jambolana,Mucuna pruriens and Tinospora cordifolia)extracts[J].Indian Journal of Experimental biology,2002,40:273-276
80 Grover JK,Vats V,Rathi SS,et al.Traditional Indian anti-diabetic plants attenuate progression of renal damage in streptozotocin-induced diabetic mice[J].Journal of Ethnopharmacology,2001,76:233-238
81 Feldman M,Schiller ER.Disorders of gastrointestinal motility associated with diabetes mellitus[J].Annals of Internal Medicine,1983,98:911-922
82 Hiller R,Sperduto RD,Ederer F.Epidemiologic associations with cataract in the 1971-1972.National Health and Nutrition Examination Survey[J].American Journal of Epidemiology,1983,118:239-249
83 Van-Heyningen R,Harding JJ.A case-control study of cataract on Ofordshire:some risk factors[J].British Journal of Opthalmology,1988,72:804-808
84 Harding JJ,Harding RS,Egerton M.Risk factors for cataract in Oxfordshire:diabetes,peripheral neuropathy,myopia,glaucoma and diarrhea[J].Acta Ophthalmolgica(Copenhagen),1989,67:517-518
85 Rathi SS,Grover JK,Vikrant V.Prevention of experimental diabetic cataract by Indian Ayurvedic plant extracts[J].Phytotherapy Research,2002b,16:774-777
86 Srivastava Y,Venkatakrishna-Bhatt H,Verma Y.Effect of Momordica charantia Linn.pomous aqueous extract on cataractogenesis in murrin alloxan diabetics[J].Pharmacol Res Commun,1988,20(3): 201-209
87 Licastro F,Franceschi C,Barbieri L,et al.Toxicity of Momordica charantia lectin and inhibitor for human normal and leukaemic lymphocytes[J].Virchows Archives of B Cell Pathology Including Molecular Pathology,1980,33:257-265
88 Ng TB,Liu WK,Sze SF,et al.Action of alpha-momorcharin,a ribosome inactivating protein,on cultured tumor cell lines[J].General Pharmacology,1994,25:75-77
89 Battelli MG,Polito L,Bolognesi A,et al.Toxicity of ribosome-inactivating proteins- containing immunotixins to a human bladder carcinoma cell line[J].International Journal of Cancer,1996,68:485-490
90 Tsao SW,Ng TB,Yeung HW.Toxicities of trichosanthin and alpha-momorcharin,abortifacient proteins from Chinese medicinal plants,on cultured tumor cell lines[J].Toxicon,1990,28:1183-1192
91 Sun Y,Huang PL,Li JJ,et al.Anti-HIV agent MAP30 modulates the expression profile of viral and cellular genes for proliferation and apoptosis in AIDS-related lymphoma cells infected with Kaposi's sarcoma-associated virus[J].Biochemical and Biophysical Research Communication,2001,287:983-994
92 Ganguly C,De S,Das S.Prevention of carcinogen-induced mouse skin papilloma by whole fruit aqueous extract of Momordica charantia[J].European Journal of Cancer Prevention,2000,9:283-288
93 Nagasawa H,Watanabe K.Inatomi H.Effects of bitter melon(Momordica charantia L.) or ginger rhizome(Zingbier offifinale rocs) on spontaneous mammary tumorigenesis in SHN mice[J].American Journal of Chinese Medicine,30:195-205
94 Claflin AJ,Vesely DL,Hudson JL,et al.,Inhibition of growth and guanylate cyclase activity of an undifferentiated prostate adenocarcinoma by an extract of the balsam pear(Momordica charantia abbreviata)[J].Proc Natl Acad Sci USA,1978,75(2):989-993
95 严启新,罗天浩.苦瓜的研究及开发思考[J].时珍国药研究,1997,8(4):380-381
96 Basch E,Gabardi S,Ulbricht C.Bitter melon(Momordica charantia):a review of efficacy and safety[J].American Journal of Health and Systemic Pharmacology,2003,65:356-359
97 Takemoto DJ,Dunford C,McMurray MM.The cytotoxic and cytostatic effects of the bitter melon on human lymphocytes[J].Toxicon,1982a,20(3):593-599
98 Takemoto DJ,Dunford C,Vaughn D,et al.Guanylate cyclase activity in human leukemic and normal lymphocytes:Enzyme inhibition and cytotoxicity of plant extracts[J].Enzyme,1982,27:179-188
99 齐文波,徐中干,徐誉泰等.苦瓜素的分离纯化与抗肿瘤活性的研究[J].离子交换及吸附,1999,15(1):59-63
100 Lee Huang S,Huang PL,Chen HC,et al.Anti-HIV and anti-tumor activities of recombinant MAP30 from bitter melon[J].Gene,1995a,61(2):151-156
101 Lee Huang S,Huang PL,Bourinbaiar,AS,et al.Inhibition of the integrase of human immunodeficiency virus(HIV) type 1 by anti-HIV plant protion MAP30 and GAP31[J].Proc Natl Acad Sci USA,1995b,92(19):8818-8822
102 Huang PL,Sun Y,Chen HC,et al.Proteolytic fragments of anti-HIV and anti-tumor proteins MAP30 and GAP31 are biologically active[J].Biochem Biophys Res Commun,1999,262:615-623
103 陈执中.抗艾滋病天然药物-苦瓜成分的研究Ⅰ,水溶性成分的分析[J].中国民族民间医药杂志,1999,37:63-65
104 Bourinbaiar AS,Lee Huang S.The activity of plant-derived antiretroviral protens MAP30 and GAP31against herpes simplex virus in vitro[J].Biochem Biophys Res Commun,1996,219(3):923-929
105 Zheng YT,Ben KL,Jin SW.Alpha-momorcharin inhibits HIV-1 replication in acutely but not chronically infected T lymphoid cells[J].Acta Pharm Sin,1999,20:239-243
106 Au TK,Collins RA,Lain TL,et al.The plant ribosome inactivating proteins luffin and saporin are potent inhibition of HIV-1 integrase[J].FEBS Letters,2000,471:169-172
107 Bourinbaiar AS,Lee-Huang S.Potentiation of anti-HIV activity of anti-inflammatory drugs,dexamethasone and indomethacin,by MAP30,the antiviral agent from bitter melon[J].Biochemistry and Biophysics Research Communication,1995,208:779-785
108 Wang HX,Ng TB.Examination of lectins,polysac charopeptide,polysaccharide,alkaloid,coumarin and trypsin inhibitors for inhibitory activity against human immunodeficiency virus reverse transcriptase and glycohydrolases[J].Planta Medica,2001a,67:669-672
109 张素,王明霞.植物药材中抗艾滋病毒活性成分研究[J].中医药信息,1994,11(4):30-33
110 Foa TL,Campadelli FG,Barbieri L,et al.Effect of ribosome-inactivating proteins on virus-infected cells:inhibition of virus multiplication and protein synthesis[J].Arch Virol,1982,71:322-332
111 Khan MR,Omoloso AD.Momordica charantia and Allium sativum:Broad spectrum antibacterial activity [J].Korean J Pharmacog,1998,29(3):155-58
112 Ogata F,Miyata T,Fujii N,et al.Purification and amino acid sequence of a bitter gourd inhibitor against an acidic amino acid-specific endopeptidase of Streptomyces griseus[J].Journal of Biological Chemistry,1991,266:16715-16721
113 Omoregbe RE,Ikuebe OM,Ihimire IG.Antimicrobial activity of some medicinal plants extracts on Escherichia coli,Salmonella paratyphi and Shigella dyseteriae[J].African Journal of Medical Science,1996,25:373-375
114 Yesilada E,Gurbuz I,Shibata H.Screening of Turkish anti-ulcerogenic folk remedies for anti-Helicobacter pylori activity[J].J Ethnopharmacol,1999,66:289-293
115 吴传茂,夏服宝,李小燕.苦瓜的抑菌作用和苦瓜茶[J].中国商办工业,2002,3:51-52
116 张绪忠,陈青山.苦瓜抑菌防腐作用的实验研究[J].预防医学情报杂志,1996,12(1):17-20
117 黎锦城.苦瓜的抗炎作用[J].中药材,2001,24(7):508-509
118 何煜波.苦瓜抗菌作用研究[J].食品科学,1998,19(3):34-36
119 陈清山,程光文,张绪忠.不同苦瓜提取液对肉类食品防腐作用的研究[J].武汉冶金科技大学学报,1996,19(3):387-390
120 陈冀胜,郑硕.中国有毒植物[M].北京:科学出版社,1987,PP.128
121 覃国芳,李远慧,高晓勤等.苦瓜果对雄性大鼠生育力的影响[J].贵阳医学院学报,1995,20(1):14-17
122 Chan WY,Tam PP,Choi HL,et al.Effects of momorcharins on the mouse embryo at the early organogenesis stage[J].Contraception,34:537-544
123 王庆华,于长春,徐誉泰等.α-苦瓜素和β-苦瓜素的研究进展[J].中草药,1995,26(5):266-267
124 Law LK,Tam PP,Yeung HW.Effects of alpha-trichosanthin and alpha-momorcharin on the development of peri-implantation mouse embryos[J].J Reprod Fert,1983,69:597-604.
125 Tam PP,Law LK,Yeung HW.Effects of alpha-momorcharin on preimplantation development in the mouse[J].J Reprod Fert,1984,71:33-38
126 常凤岗,李建梅.苦瓜抗生育活性成分的化学研究(Ⅰ)[J].中草药,1995,21(6):281-284
127 Schreiber CA,Wan L,Sun Y,et al.The antiviral agents,MAP30 and GAP31,are no toxic to human spermatozoa and may be useful in preventing the sexual transmission of human immunodeficiency virus type 1[J].Fertil Steril,1999,72(4):686-690
128 乔昕,陈雄,李冬生.苦瓜抗氧化作用的初步研究[J].湖北农业科学,2003,1:74-75
129 傅明辉.苦瓜籽蛋白的分离、纯化及其抗氧化作用的研究[J].药物生物技术,2001,8(5):248-250
130 傅明辉,陈健汉,庄东红.苦瓜提取液的抗氧化、抑菌和降血糖活性[J].食品科学,2001,22(4):88-90
131 Leung SO,Yeung HW,Leung KN.The immunosuppressive activities of two abortifacient proteins isolated from the seeds of bitter melon(Momordica charantia)[J].Immunopharmacology,1987,13(3):159-171
132 Spreafico F,Malfiore C,Moras ML.The immunomodulatory activity of the plant proteins Momordica charantia inhibitor and pokeweed antiviral protein[J].International Journal of Immunopharmacology,1983,5:335-343
133 Cunnick JE,Sakamoto K,Chapes SK.Induction of tumor cytotoxic immune cells using a protein from the bitter melon(Momordica charantia)[J].Cellular Immunology,1990,126:278-289
134 程光文,陈清山,张绪忠.苦瓜对正常小鼠和免疫受抑小鼠免疫促进作用的实验观察[J].中国冶金工业医学杂志,1996,13(4):198-200
135 Sitasawad SL,Shewade Y,Bhonde R.Role of bittergourd fruit juice in stz-induced diabetic state in vivo and in vitro[J].J Ethnopharmacol,2000,73(1-2):71-79
136 Welihinda J,Arvidson G,Gylfe E,et al.The insulin-releasing activity of the tropical plant Momordica charantia[J].Acta Biot Mel Ger,1982,41(12):1229-1240
137 Shibib BA,Khan LA,Rahan R.Hypoglycaemic activity of Coccinia indica and Momordica charantia on diabetic rats:depression of the hepatic gluconeogenic enzymes glucose-6-phosphatase and fructose-1,6-biphosphatase and elevation of both liver and red-cell shunt enzyme glucose-6-phosphate dehydrogenase[J].Biochemistry Journal,1993,292:267-290
138 袁宝君,戴月.癞葡萄胶囊调节血糖人体试食试验的研究[J].卫生研究,2003,32:326-328
139 石根勇,吕中明,陈新霞等.癞葡萄冻干粉凋节血糖作用的研究[J].中国生化药物杂志,2005,26(5):303-304
140 冯怀蓉,张慧涛,茆军.多肽简介及应用[J].新疆农业科学,2002,39(1):38-39
141 冯秀燕,计成.寡肽在蛋白质营养中的作用[J].动物一样学报,2001,13(3):8-13
142 Webb KE Jr,DiRienzo DB,Mathews JC.Recent development in gastrointestinal absorption and tissue utilization of peptides:a review[J].J Dairy Sci,1993,76:351-361
143 Gobbetti M,Stepaniak L,de Angelis M.Latent bioactive peptides in milk proteins:proteolytic activation and significance in dairy processing[J].Cri Rev Food Sci Nutri,2002,42(3):223-239
144 Vincenzini MT,Iantomasi T,Favilli F.Glutathione transport across intestinal brush-border membranes:effects of ions,pH,delta psi,and inhibitors[J].Biochim Biophys Acta,1989,987(1):29-37
145 Dantzing et al.Association of intestinal peptide transport with a protein related to the cadheririn super family[J].Science,1994,764:430-433
146 蒲首丞,王金水.生物活性肽的制备与应用[J],2005,5:49-51
147 刘平,田桂玲,叶蕴华.固载化酶催化合成多肽的研究进展[J].高等学校化学学报,2001,8:1342-1348
148 Meisel H.Biochemical properties of regulator peptides derived from milk proteins[J].Biopolymers,1997,43(2):119-125
149 Meisel H,Bockelmann W.Bioactive peptides encrypted in milk proteins proteolytic activation and thropho-functional properties[J].Antonie Van Leeuwenhoek,1999,76:207-213
150 Gill I,Lopez-Fandino R,Jorba X.Biologically active peptides and enzymatic approaches to their production[J].Enzyme and Microbial Technology,1996,18:162-183
151 Korhonen H,Pihlanto A.Food-derived bioactive peptides-opportunities for designing future foods[J].Current Pharmaceutical Design,2003,9:1297-1308
152 邵丙扬.警惕世界面临糖尿病灾难[J].内科急危重症杂志,2004,10(1):1-2
153 Zhang BB,Moller DE.News approaches in the treatment of type 2 diabetes[J].Curr Opin Chem Biol,2000,4:461-467
154 Tolman KG,Chandramouli J.Hepatotoxicity of the thiazolidinediones[J].Clin Liver Dis,2003,7:369-370.
155 王吉影,顾耀,徐乃馨等.阿卡波糖和伏格列波糖治疗2型糖尿病的比较[J].同济大学学报(医学版),2001,2(21):32-35
156 Delea TE,Edelsberg JS,Hagiwara M,et al.Use of thiazolidinediones and risk of heart failure in people with type 2 diabetes:a retrospective cohort study[J].Diabetes Care,2003,26:2983-2989
157 张冬英.普洱茶降糖降脂活性成分的研究[D].博士学位论文,湖南农业大学,2006
158 Yoshikawa M,Fujita H,Matoba N,et al.Bioactive peptides derived from food protein preventing lifestyle-related diseases[J].Biofactors,2000,12:143-146.
159 Dziuba J,Minkiewicz P,Nalecz D.Biologically active peptides derived from plant and animal proteins [J].Pol Food Nutri Sci,1999,8:3-16
1 范冠杰主编.糖尿病[M].北京:人民卫生出版社,2002
2 Piedrola G,Novo E,Escober F,et al.White blood cell count and insulin resistance in patients with coronary artery disease[J].Annual Endocrinology(Paris),2001,62:7-10
3 Yaryur-Tobias JA,Pinto A,Neziroglu F.Anorexia nervosa,diabetes mellitus,brain atrophy and fatty liver [J].International of Etiological Disorders,2001,30:350-353
4 Akhtar MS,Iqbal J.Evaluation of the hypoglycemic effect of Achyranthes aspera in normal and alloxan diabetic rabbits[J].J Ethnopharmacol,1991,31:49-57
5 Tolman KG,Chandramouli J.Hepatotoxicity of the thiazolidinediones[J].Clinics in Liver Disease,2003,7:369-370
6 Zhang BB,Moller DE.News approaches in the treatment of type 2 diabetes[J].Curr Opin Chem Biol,2000,4:461-467
7 Li WL,Zheng HC,Bukuru J,et al.Natural medicines used in the traditional Chinese medical system for therapy of diabetes mellitus[J].J Ethnopharmacol,2004,92:1-21
8 Ugochukwu NH,Babady NE.Antihyperglycemic effect of aqueous and ethanolic extracts of Gongronema latifolium leaves on glucose and glycogen metabolism in livers of normal and streptozotocin-induced diabetic mice[J].Life Sci,2003,73:1925-1938
9 Eddouks M,Lemhadri A,Zeggwagh NA,et al.Potent hypoglycaemic activity of the aqueous extract of Chamaemelum nobile in normal and streptozotocin-induced diabetic rats[J].Diabetes Res Clin Pract,2005,67(3):189-195
10 Zhang RX,Zhou JH,Jia ZP,et al.Hypoglycemic effect of Rehmannia glutinosa oligosaccharide in hyperglycemic and alloxan-induced diabetic rats and its mechanism[J].J Ethnopharmacol,2004,90:39-43
11 Sezik E,Aslan M,Yesilada E,et al.Hypoglycaemic activity of Gentiana olivieri and isolation of the active constituent through bioassay-directed fractionation techniques[J].Life Sci,2005,76:1223-1238
12 Lee MK,Kim MJ,Cho SY,et al.Hypoglycemic effect of Du-zhong(Eucommia ulmoides Oliv.) leaves in streptozotocin-induced diabetic rats[J].Diabetes Res Clin Pract,2005,67:22-28
13 Roy S,Sehgal R,Padhy BM,et al.Antioxidant and protective effect of latex of Calotropis procera against alloxan-induced diabetes in rats[J].J Ethnopharmacol,2005,102:470-473
14 Joy KL,Kuttan R.Anti-diabetic activity of Picrorrhiza Kurroa extract[J].J Ethnopharmacol,1999,167:143-148
15 Yadav S,Vats V,Dhunnoo Y,et al.Hypoglycemic and antihyperglycemic activity of Murraya koenigii leaves in diabetic rats[J].J Ethnopharmacol,2002,82:111-116
16 Ali L,Khan AK,Mamun MI,et al.Studies on hypoglycemicin effects of fruit pulp seed and whole plant of Momordia charantia on normal and diabetic model rats[J].Planta Med,1993,59:408-412
17 Jouad H,Eddouks M,Lacaille-Dubois MA.Hypoglycaemic effect of the water extract of Spergularia purpurea in normal and streptozotocin-induced diabetic rats[J].J Ethnopharmacol,2000,71:169-177
18 李聪然,游雪甫,蒋建东.糖尿病动物模型及研究进展[J].中国比较医学杂志,2005:15(1):59-63
19 黄伟坤.食品检验与分析[M].北京:中国轻工业出版社,1997
20 赵亚华,高向阳.生物化学实验技术教程[M].广州:华南理工大学出版社,2000
21 郑建仙.功能性食品[M].北京:中国轻工业出版社,1999,486-487
22 吴有炜.试验设计与数据处理[M].苏州:苏州大学出版社,2002
23 陈晓超.苦瓜提取物中降糖物质的分离鉴定及其降糖机理的初步探讨[D].福州大学,硕士学位论文,2004
24 http://sun.ars-grin.gov:8080/npgspub/xsql/duke/plantdisp.xsql?taxon=694
25 http://sun.ars-grin.gov:8080/npgspub/xsql/duke/plantdisp.xsql?taxon=843
26 Takasu N,Komiya I,Asawa T,et al.Streptozocin and alloxan-induced H_2O_2 generation and DNA fragmentation in pancreatic islets[J].Diabetes,1991,40(9):1141-1145
27 潘竞锵,韩超,刘惠纯等.菌陈蒿汤对正常和多种糖尿病模型动物血糖的影响[J].中药材,2001, 24(2):128-131
28 蒋渝,周永禄.四氧嘧啶诱发糖尿病模型的建立[J].上海实验动物科学,]990,10(2):79-81
29 张传仓,刘卫鹏,杜江等.改良四氧嘧啶法制作糖尿病动物模型[J].实用医学杂志,2007,23(5):623-624
30 Utkan T,Yildirim MK,Yildirim S,et al.Effects of the specific phosphodiesterase inhibitors on alloxan-induced diabetic rabbit cavernous tissue in vitro[J].Int J Impot Res,2001,13(1):24-30
31 Vats V,Grover JK,Rathi SS.Evaluation of anti-hyperglycemic and hypoglycemic effect of Trigonella foenum-graecum Linn,Ocimum Sanctum Linn and Pterocarpus marsupium Linn,in normal and alloxanized diabetic rats[J].J Ethnopharmacol,2002,79:95-100.
32 金丕焕.医用统计方法(第二版)[M].上海:复旦大学出版社,2003
33 冯礼明.单一自由度比较分析方法介纠[J].数理统计与管理,1990,1:21-26
34 周乃健,赵志立.单一自由度分组比较统计分析方法在农业试验中的应用[J].山西农业大学学报(自然科学版),1992,12(2):110-114
35 Sitasaward SL,Shewade Y,Bhonde R.Role of bittergourd fruit juice in STZ-induced diabetic state in vivo and in vitro[J].J Ethnopharmacol,2000,73:71-79
36 Chattopadhyay RR,Sarkar SK,Ganguly S,et al.Hypoglycemic and antidiabetic effects of leaves of Vinca rosea Linn[J].Indian Journal of Physiology and Pharmacology,1991,35:145-151
37 Sharma SR,Dwivedi SK,Swarup D.Hypoglycaemic and hypolipidemic effects of Cinnamomum tamala Nees leaves[J].Indian Journal of Experimental Biology,1996,34:372-374
38 Rao VV,Dwivedi SK,Swarup D,et al.Hypoglycemic and antidiabetic effects of Aegle marmelose leaves in rabbits[J].Current Science,1995,69:932-933
39 张芳林,李果,刘优萍等.2型糖尿病大鼠模型的建立及其糖代谢特征分析[J].中国实验动物学报,2002,10(1):16-20
40 池芝盛主编.糖尿病学(第1版)[M].北京:人民卫生出版社,1986:161-162
1 汪家政,范明主编.蛋白质技术手册[M].北京:科学技术出版社,2000
2 赵锐,顾谦群,管华师.多肽物质分离与分析方法研究进展[J].中国海洋药物,2000,3:48-53
3 Nebot VS,Benavente F,Hernadex E,et al.Evaluation of the electrophoretic behaviour of opioid peptides separation by capillary electrophoresis-electrospray ionzation mass spectrometry[J].Analytical Chimmica Acta,2006,577:68-76
4 Herraiz T.Sample preparation and reversed phase-high performance liquid chromatography analysis of food-derived peptides[J].Analytica Chimica Acta,1997,352:119-139
5 Bayerbach R,Nguyen VD,Schurr U,et al.Characterization of the water-insoluble fraction from fast pyrolysis liquids(pyrolytic lignin) part Ⅲ.Molar mass characteristics by SEC,MALDI-TOF-MS,LDI-TOF-MS,and Py-FIMS.J Anal Appl Pyrolysis,2006,77(2):95-101
6 Sporns P,Wang J.Exploring new frontiers in food analysis using MALDI-MS[J].Food Res Int,1998,31(3):181-189
7 时钧、袁权.膜技术分离手册[M].北京:化学工业出版社,2001
8 刘茉娥.膜分离技术[M].北京:化学工业出版社会,1998
9 Rautenback R,Albrecht R.Membrane Process[M].Chichester:John Wiley and Sons Ltd,1989,pp.2-15
10 Kawakatsu T,Kobayashi T,Sano Y,et al.Clarification of green tea extract by microfiltration and ultrafiltration[J].Biosci Biotech Biochem,1995,59(6):1016-1020
11 张晓楠,郭立安.超滤在蛋白质纯化中的应用[J].中国生化药物杂志,1999,20(2):97-98
12 Alvise ND,Lesueur-Lambert C,Fertin B.Hydrolysis and large scale ultrafiltration study of alfalfa protein concentrate enzymatic hydrolysate[J].Enzyme and Microbial Tehcnology,2002,27:286-294
13 刘忠洲,续曙光,李锁定.微滤、超滤过程中的膜污染与清洗[J].水处理技术,1997,23(4):187-193
14 郑成.膜的清洗及杀菌[J].化工进展,1996,6:45-49
15 邬行彦,孙进.膜过滤法及其在抗生素工业中的应用[J].国外医药抗生素杂志分册,1992,13(6):455-462;1993,14(1):60-67
16 刘茉娥.超滤膜污染机理的研究与控制[J].水处理技术,1989,3:163-169
17 张赞红,施亚钧.超滤膜分离技术的应用及膜污染问题[J].化学世界,1993,6:242-245
18 任建新.膜分离技术及其应用[M].北京:化学工业出版社,2003:1
19 华耀祖.超滤技术与应用[M].北京:化学工业出版社工业装备与信息工程出版中心,2004
20 Rao A,Shallo HE,Ericson HE,et al.Characterization of soy protein concentrate produced by membrane ultrafiltration[J].Journal of Food Science,2002,67(4):1412-1418
21 Shallo HE,Rao A,Ericson AP,et al.Preparation of soy protein concentrate by ultrafiltration[J].Journal of Food Science,2001,66(2):242-246
22 龙晓丽.红三鱼蛋白水解物的制备及分离技术的研究[D].硕士学位论文,华南理工大学,2004
23 江华.超滤及冷冻浓缩技术在低聚木糖制备中的应用[D].博士学位论文,南京林业大学,2001
24 赵永芳.生物化学技术原理及其应用[M].武汉大学出版社,1997
25 Irvine GB.High-performance size-exclusion chromatography of peptides[J].J Biochem Biophys Methods,2003,56:233-242
26 韩冬,叶美玲,施良和.水溶性凝胶色谱中的非体积排阻效应[J].色谱,1995,13(6):432-436
27 Dan N,Ganesan R,Flood KG,et al.Determination of enantiomers in a synthetic argininal peptide using capillary zone electrophoresis and high-performance liguid chromatography[J].J Chromatogr,2000,891:115-120
28 Sanz NV,Benavente F,Toro I,et al.Optimization of HPLC conditions for the separation of complex crude mixtures produced in the synthesis of therapeutic peptide hormones[J].Chromatogr,2000,891:115-120
29 Smith DD,Saha S,Fang G,et al.Modifications to the N-terminus but not the C-terminus of calcitonin gene-related peptide(8-37) produce antagonists with increased affinity[J].J Med Chem,2003,46:2427-2435
30 Purcell AW,Zhao GL,Aguilar MI,et al.Comparison between the isocratic and gradient retention behavior of polypeptides in reversed-phase liquid chromatographic environments[J].J Chromatogr,1999,852:43-46
31 王英武,王玲,顾景凯等.电喷雾-串联四极杆-飞行时间质谱法分析寡肽的一级结构[J].分析化学,2003,31(6):709-712
32 邓慧敏,赖志辉,黎军等.碎片结构分析在MALDI-TOF-MS法测定多肽序列中的应用[J].生物化学与生物物理学报,2000,32(2):179-182
33 Medzihradszky KF,Adems GW,Burlingame AL,et al.Peptide sequence determination by matrix-assisted laser desorption ionization employing a tandem double focusing magnetic orthogonal acceleration time-of-flight mass spectrometry[J].J Am Soc Mass Spetrom,1996,7:1-10
34 Kaufmann R.Maxtrix-assisted laser desorption ionization(MALDI) mass spectrometry:a novel analytical tool in molecular biology and biotechnology[J].J Biotechnol,1995,41:155-175
35 卞则梁,王光辉.质谱与生命科学[J].化学通服,1994,7:40-46
36 桑志红,杨松成.电喷雾电离质谱及其在质白质化学研究中的应用[J].国外医学药学分册,2000,27(1):38-41
37 梁宋平.世纪之交的蛋白质序列测定技术[J].生命科学,1999,1:31-34
1 Yoshikawa M,Fujita H,Matoba N,et al.Bioactive peptides derived from food protein preventing lifestyle-related diseases[J].Biofactors,2000,12:143-146.
2 Dziuba J,Minkiewicz P,Nalecz D.Biologically active peptides derived from plant and animal proteins [J].Pol Food Nutri Sci,1999,8:3-16
3 Suetsuna K,Chen JR.Isolation and characterization of peptides with antioxidant activity bioprocesses used in isolation and recovery[J].Curr Pharm Des,2003,9(16):1309-1323
4 Mahmoud MI.Physicochemical and functional properties of protein hydrolysates in nutritional products [J].Food Technol,1994,48:89-95
5 夏书华.螺蛳中生物活性物质的研究[D].博士学位论文,江南大学,2006
6 孔祥珍.小麦面筋蛋白的酶解及其阿片肽的研究[D].博士学位论文,江南大学,2006
7 Suetsuna K,Chen JR.Isolation and characterization of peptides with antioxidant activity derived from wheat gluten[J].Food Science and Technology Research,2002,8(3):227-230
8 Schusdziarra V,Schick R,Holand A,et al.Effect of opiate-active substances on pancreatic polypeptide levels in dogs[J].Peptides,1983,4:205-210
9 Nissen JA.Enzymatic hydrolysis of food protein.New York:Elsevier Applied Science Pulisher,1986
10 黄伟坤.食品检验与分析[M].北京:中国轻工业出版社,1990
11 李朝兴,印寿根,王克雷等.大孔吸附树脂对绞股蓝皂甙的吸附研究[J].离子交换与吸附,1994,10(3):203-207
12 赵亚华.生物化学实验技术教程[M].广州:华南理工大学出版社,2000
13 王章存,姚惠源.大米蛋白质的酶法水解及其性质研究[J].中国粮油学报,2003,18(5):5-7
14 程云辉.麦胚蛋白酶解物的制备、结构及其生物活性功能的研究[D].博士学位论文,江南大学,2006
15 Marquez MC,Vazquez MA.Modeling of enzymatic protein hydrolysis[J].Process Biochem,1999,35:111-117
16齐崴,何志敏,何丽霞.酪蛋白酶解制备酪蛋白磷酸肽研究Ⅰ过程分析与优化[J].食品科学,2001,22(7):25-28
17 Godfrey T,Reichelt J.Industrial Enzymology:the application of enzymes in industry.1983.England,Macmillan Publishers Ltd.pp 539-540,497-498
18王璋编.食品酶学[M].北京,轻工业出版社,1990,pp 180-185
19 Adamson NJ,Reynolds EC.Characterization of casein phosphopeptides prepared using Alcalase:determination of enzyme specificity[J].Enzyme and microbial technology,1996,19:202-207
20 Triveni R,Shamala TR,Rastogi NK.Optimised production and utilization of exopolysaccharide from Agrobacterium radiobacter[J].Process Biochemistry,2001,36:787-795
21 Kilic M,Bayraktar E,Ates S.Investigation of extractive citric acid fermentation using response-surface methodology[J].Process Biochemistry,2002,37:759-767
22 Mizubuti IY,Biondo Juniora O,de Oliveira Souza LW,et al.Response surface methodology for extraction optimization of pigeon pea protein[J].Food Chemistry,2000,70:259-265
23 Nag B,Medicherla S,Sharma SD.Orally active fraction of Momordica charantia,active peptides thereof,and their use in the treatment of diabetes[P].U.S.(P) US6127338,2000-10-03
24 Khanna P,Jain SC.Panangariya A.Hyopglycemic activity of polypeotide-P from a plant source[J].Nat Prod,1981,44:648-655
25 Owen R.Fennema著,王璋等译.食品化学(第三版)[M].北京:中国轻工业出版社,2003
26 邓劲光,胡熙恩,朱永.大孔吸附树脂对氢化可的松的吸附与洗脱性能研究[J].离子交换与吸附,2000,16(2):134-139
1 Charpentier G.Oral combination therapy from type 2 diabetes[J].Diabetes/Metabolism Research and Reviews,2002,18(Suppl 3):$70-$76
2 Ahmed I,Adeghate E,Sharma AK,et al.Effects of Momordica charantia fruit juice on islet morphology in the pancreas of the streptozotocin-diabetic rat[J].Diabetes Research Clinical Practices,1998,40:145-151
3 Welihinda J,Arvidson G,Gylfe E,et al.The insulin-releasing activity of the tropical plant Momordica charantia[J].Acta Biot Mel Ger,1982,41(12):1229-1240
4 Javill-Taylor KJ,Anderson RA,Graves DJ.A hydroxychalcome derived from cinnamon functions as a mimetic for insulin in 3T3-L1 adipocytes[J].Journal of the American College of Nutrion,2001,20:327-336
5 Qin B,Nagasaki M,Ren M,et al.Cinnamon extract(traditional herb) potentiates in vivo insulin-regulated glucose utilization via enhancing insulin signaling in rat[J].Diabetes Research and Clinical Practice,2003,63:139-148
6 Zhang RX,Zhou JH,Jia,ZP,et al.Hypoglycemic effect of Rehmannia glutinosa oligosaccharide in hyperglycemic and alloxan-induced diabetic rats and its mechanism[J].Journal of Ethnopharmacology,2004,90:39-43
7 沈同,王镜岩.生物化学[M].北京:高等教育出版社,1999
8 郑建仙.功能性食品[M].北京:中国轻工业出版社,1999,pp.486-487
9 Gold AH.The effect of diabetes and insulin on liver glycogen synthetase activation[J].Journal of Biological Chemistry,1970,245:903-905
10 Goel RK,Mahajan MP,Kulkarni SK.Evaluation of anti-hyperglycemic activity of some novel monocyclic beta lactams[J].J Pharm Pharm Sci,2004,7(1):80-83
11 Lolitkar MM,Rao MRR.Pharmacology of hypoglycaemic principle isolated from the fruits of Charantia Linn[J].Indian Journal Pharmacy,1966,28(5):129-133
12 Sharma SB,Nasir A,Prabhu KM,et al.Hypoglycaemic and hypolipidemic effect of ethanolic extract of seeds of Eugenia jambolana in alloxan-induced diabetic rabbits[J].Journal of Ethnopharmacology,2003,85:201-206
13关子安,孙茂欣,关大顺等.现代糖尿病学[M].天津:天津科学技术出版社,2000
14黄燕飞,刘志红.脂毒性与2型糖尿病及其并发症的关系[J].肾脏病与透析肾移植杂志,2002,11(1):58-63
15 Ginsberg HN.Basic mechanisms underlying the common hypertriglyceridemia and low HDL cholesterol levels[J].Diabetes,1996,45:S27-S30
16刘静,吴兆苏,赵冬.白由脂肪酸与代谢综合症的关系[J].心肺血管杂志,2005,24:125-127
17 Dhandapani S,Subramanian VR,Rajagopal S,et al.Hypolipidemic effect of Cuminum Cyminum L.on alloxan-induced diabetic rats[J].Pharmacological Research,2002,46(3):251-255
18 Bergman RN,Ader M.Free fatty acids and pathogenesis of type 2 diabetes mellitus[J].Trends Endocrinol Metab.2000.11:351-356
19邹效漫.二型糖尿病的脂蛋白代谢特点[A].见:叶平.血脂的基础与临床[M].北京:人民军医出版社,2002,191-200
20朱姿英,薛耀明.高甘油三酯血症和2型糖尿病[J].中国糖尿病杂志,2003,11(2):153-154
21 许樟荣,王爱红.糖尿病与血脂异常[J].中华老年医学杂志,2006,25(4)
22 Bopanna KN,Kannan J,Gadgil S,et al.Antidiabetic and antihyperglycemic effects of neem seed kernel powder on alloxan diabetic rabbits[J].Indian Journal of Pharmacology 1997,29:162-167
23 Luc G,Fruchart JC.Oxidation of lipoproteins and atherosclerosis.American Journal of Clinical Nutrition,1991,53:2065-2095
24 Ram A,Lauria P,Gupta R,et al.Hypocholesterolemic effects of Teminalia arjuna tree bark[J].Journal of Ethnopharmacology,1997,55:165-169
25 Sharma SR,Dwivedi SK,Swarup D.Hypoglycaemic,anti-hyperglycaemic and hypolipidemic activities of Caesalpinia sylvestre seeds in rats[J].Journal of Ethnopharmacology,1997,58:39-44
26 Jouad H,Lemhadri A,Maghrani M,et al.Cholesterol lowering activity of the aqueous extract of Spergularia in normal and recent onset diabetic rats[J].Journal of Ethnopharmacology,2003,87:43-49
27 Oberley LW.Free radicals and diabetes[J].Free Radical Biol and Med,1988,5:13 - 24
28 Jemings PE.The relationship of oxidative stress to thrombotic in type Ⅰ diabetic patients with diabetic retinopathy[J].Diabetic Med,1992,87:860 - 865
29 Maxwell SRJ,Thomason S,Sandier D,et al.Antioxidant status in patients with uncomplicated insulin-dependent and non-insulin-dependent diabetes mellitus[J].European Journal of Clinical Investigation,1997,27:484-490
30 Zhang XF,Tan BKH.Antihyperglycemic and antioxidant properties of Andrographis paniculata in normal and diabetic rats[J].Clinical and Experimental Pharmacology and Physiology,2000,27:358-363
31 El-Missiry MA,Othman AI,Amer MA.L-Arginine ameliorates oxidative stress in ailoxan-induced experimental diabetes mellitus[J].Journal of Applied Toxicology,2004,24:93-97
32 Manonmani G,Bhavapriya V,Kalpana S,et al.Antioxidant activity of Cassia Fistula(linn.) flowers in alloxan induced diabetic rats[J].Journal of Ethnopharmacology,2005,97:39-42
33 金晖,唐尧.糖尿病中氧化物与抗氧化物损伤[J].医学综述,1996,2(2):49-51
34 Vassalle C,Lubrano V,Abbate A,et al.Determination of metric plus nitrate and malondialdehyde in human plasma:analytical performance and the effect of smoking and exercise[J].Clin Chem Lab Med,2002,22(2):130-134
35 陈瑗.自由基医学基础与病理生理(第一版)[M].北京:人民卫生出版社,2002
36 丁爱玲,杨鄂平,吴丽霞等.中药对运动大鼠肾脏和肌肉自由基代谢的影响[J].北京体育大学学报,2004,27(2):206-208
37 刘方,武子斌,牛淑敏等.中药材抗氧化及自由基清除活性的研究[M].中国药学杂志,2001,36(7):442-445
38 Park SY,Bok SH,Jeon S,et al.Effect ofrutin and tannic acid supplements on cholesterol metabolism in rats[J].Nutrition Research,2002,22:283-295
39 Ohkawa H,Ohishi N,Yake K.Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction [J].Anal Biochem,1979,95:351-358
40 Chugh SN,Dhawan R,Kishore K,et al.Glibenclamide vs.gliclazide in reducing oxidative stress in patients of noninsulin dependent diabetes mellitus - a double blind randomized study[J].Journal of Association of Physicians of India,2001,49:803-807
41 Elmali E,Altan N,Bukan N.Effect of the sulphonylurea glibenclamide on liver and kidney antioxidant enzymes in streptozocin-induced diabetic rats[J].Drugs in R & D,2004,5(4):203-208
42 Asayama K,Nfeler F,Eenglish D,et al.Alloxan-induced free radical production in isolated cells,selective effect on islets cells[J].Diabetes,1984,33(10):1008-1011
43 蔡英兰,金政,金香子,等.榛花对糖尿病小鼠降糖作用及对肝脏和胰岛病理学影响[J].辽宁中医杂志,2002,32(12):1325-1326
2 江苏新医学院.中药大辞典[M].上海:上海科学技术出版社,1980,1289-1290
3 程霜,崔庆新,王勇.水溶性苦瓜多糖的提取与鉴定[J].郑州粮食学院学报,2000,21(2):53-56
4(明)李时珍.本草纲目[M].北京:人民卫生出版社,1982
5 田亚维.苦瓜的研究概述[J].医学信息,2002,15(10):619-670
6 邓向军,徐斌,董英.苦瓜化学成分的研究进展[J].时珍国医国药,2006,17(12):2449-2451
7 Okabe H,Miyahara Y,Yamauchi T,et al.Studies on the constituent of Momordica charantia L.isolation and characterization of Momordicosides A and B[J].Chem Pharm Bull,1980,28:2753-2762
8 Miyahara Y,Okabe H,Yamauchi T.Studies on the Constituent of Momordica charantia L.Ⅱ.Isolation and Characterization of minor seed glycosides,Momordicosides C,D and E[J].Chem Pharm Bull,1981,29(6):1561-1566
9 Yasuda M,Iwamoto M,Okabe H,et al.Structures of momordicines Ⅰ,Ⅱand Ⅲ:the bitter principles in the leaves and vines of Momordica charantia L[J].Chem Pharm Bul,1984,32(5):2044-2047
10 Murakami T,Emoto A,Matsuda H,et al.Structures of New Cucurbitane-Type Triterpene Glycosides,Goyaglycosides-a,-b,-c,-d,-e,-f,-g,and -h,and New Oleanane-Type Triterpene Saponins,Goyasaponins Ⅰ,Ⅱ,and Ⅲ,from the fresh fruit of Japanese Momordica charantia L.[J].Pharmaceutical Society of Japan,2001,49(1):54-63
11 Raman A,Lau C.Anti-diabetic properties and phytochemistry of Momordica charantia L.(Cucurbitaceae)[J].Phytomedicine,1996,2(4):349-362
12 Guevara AP,Linn Sylianco C,Dayrit F,et al.Antimutagens from Momordica charantia[J]Mutat Res,1990,230(2):120-123
13 Sucrow W.Uber steringlucoside und ein neues stigmastadienol aus Momordica charantia[J].Tetrahedron Lette,1965,26(9):2217
14 Lotlikar MM,Rajarama RMR.Pharmacology of a hypoglycemic principle:isolated from the fruits of Momordica charantia Linn[J].Indian Journal of Pharmacy,1966,28:129-133
15 Khanna P,Jain C,Bansal R.Effect of cholesterol on growth and production of diosgenin,gitogenin,titogenin and sterols in suspension cultures[J].Indian J Exp Biol,1975,13:211-213
16 Lau C,Raman A,Noel M,et al.Phytochemicals isolated from the anti-hyperglycaemic hexane extract of the unripe fruit of Momordica charantia L[J].J Pharm Pharmacol Suppl,1998,50:84
17 常凤岗.苦瓜的化学成分研究[J].中草药,1995,26(10):507-509
18 Khanna P,Jain SC.Panangariya A.Hyopglycemic activity of polypeotide-P from a plant source[J].Nat Prod,1981,44:648-655
19 Mokkhasmit M,Swatdimongkol K,Satrawaha P.Study on toxicity of Thai medicinal plants[J].Bull Dept Med Sci,1971,12(2/4):36-65
20 Nag B,Medicherla S,Sharma,SD.Orally active fraction of Momordica charantia,active peptides thereof,and their use in the treatment of diabetes[P].U.S.(P) US6127338,2000-10-03
21 Tomita M,Kurokawa T,Onozaki K,et al.Purification of galactose-binding phytoagglutinins and phytotoxin by affinity column chromatography using sepharose[J].Experientia,1972,28(1):84-85
22 Montecucchi PC,Lazzarin AM,Barbieri L,et al.N-termial sequence of some ribosome inactivating protein[J].Int J Pept Protein Res,1989,33:263-267
23 Kubota S,Yeung HW,Yang JT.Conformation of abortifacient proteins:trichosanthin,α-momorcharin and β-momorcharin[J].Biochemica et biophysica acta,1986,871:101-106
24 Pu Z,Lu BY,Liu WY,et al.Characterization of the enzymatic mechanism of gamma-momorcharin,a novel ribosome-inactivating protein with lower mocular weight of 11,5000 purified from the seeds of bitter gourd[J].Biochem Biophys Res Commun,1996,229(1):287-294
25 Barbieri L.Inhibition of protein synthesis in vitro by proteins from the seeds of Momoridca charantia[J].Biocetem J,1980,186:443-452
26 Lee-Huang S,Huang PL,Nara PL.MAP 30:a new inhibitor of HIV-1 infection and replication[J].FEBS lett,1990,272:12-18
27 Jiratchariyakul W,Wiwat C,Vongsakul M,et al.HIV inhibitor from Thai bitter gourd[J].Planta Med,2001,67(4):350-353
28 彭爱芝.苦瓜中氨基酸和无机元素的含量分析[J].湖南医科大学学报,1996,21(4):305
29 于长春,郑长春,王庆华等.苦瓜籽中脂肪酸的分离及生理作用研究[J].松辽学刊(自然科学版),1995,2:40-41
30 El-Gengaihi S,Karawya MS,Selim MA,et al.A novel pyrimidine glycoside from Momordica charantia L.[J].Pharmazie,1995,50(5):361-362
31 郭亦然,王水,唐琳等.苦瓜籽的生药学研究[J].四川大学学报(自然科学版),2001,38(3):454-456
32 李梦华,商学义.苦瓜对大白鼠血糖的影响[J].天津师大学报(自然科学版),1992,20(2):52-54
33 Akhtar S,Akhtar,AM,Yaqub M.Effect of Momordica charantia on blood glucose level of normal and alloxan-diabetic rabbits[J].Planta Medica,1981,42(3):205-212
34 Hang YY,Zhou YF,Chen ZM,et al.Pharmacological experiments of freeze-dried powder of balsam pear (Momordica charantia L.) on lowering the blood glucose level to mice and its acute toxicity[J].J Plant Resour & Environ,2000,9:19-21
35 黄小琴,杨延莉.苦瓜胶囊降血糖作用研究[J].药学服务与研究,2002,2(2):84-85
36 Reyes BA,Bautista ND,Tanquilut NC,et al.Anti-diabetic potentials of Momordica charantia and Andrographis paniculata and their effects on estrous cyclicity of alloxan-induced diabetic rats[J].J Ethnopharmacol,2006,105(1-2):196-200
37 Karunanayake EH,Jeevathayaparan S,Tennekoon KH,et al.Effect of Momordica charantia fruit juice on the streptozotocin-induced diabetic rats[J].J Ethnopharmacol,1990,30(2):199-204
38 Ahmed I,Adeghate E,Sharma AK,et al.Effects of Momordica charantia fruit juice on islet morphology in the pancreas of the streptozotocin-diabetic rat[J].Diabetes Research Clinical Practices,1998,40:145-151
39 Ahmed I,Lakhani MS,Gillett M,et al.Hypotriglyceridemic and hypocholesterolemic effects of anti-diabetic Momordica charantia(karela) fruit extract in streptozotocin-induced diabetic rats[J].Diabetes Research Clinical Practices,2001,51:155-161
40 Shetty AK,Kumar GS,Sambaiah K,et al.Effect of bitter gourd(Momordica charantia) on glycaemic status in streptozotocin induced diabetic rats[J].Plant Foods Hum Nutr,2005,60(3):109-112
41 Ojewole JA,Adewole SO,Olayiwola G,et al.Hypoglycaemic and hypotensive effects of Momordica charantia Linn(Cucurbitaceae) whole-plant aqueous extract in rats[J].Cardiovasc J S Afr,2006,17(5):227-32
42 Liaquat Ali,Abul Kalam Azad Khan,Muhammad lqbal Rouf Mamun,et al.Studies on hypoglycemic effects of fruit pulp,seed and whole plant of Momordica charantia on normal and diabetic model rats[J].Planta Med,1993,59:408-412
43 Kumar Shetty A,Suresh Kumar G,Veerayya Salimath P,et al.Bitter gourd(Momordica charantia)modulates activities of intestinal and renal disaccharidases in streptozotocin-induced diabetic rats[J].Mol Nutr Food Res,2005,49(8):791-796
44 Jayasooriya AP,Sakono M,Yukizaki C,et al.Effects of Momordica charantia power on serum glucose levels and various lipid parameters in rats fed with cholesterol-free and cholesterol-enriched diets[J].J Ethnopharmacol,2000,72:331-336
45 Miura T,Itoh C,Iwamoto N,et al.Hypoglycemic activity of the fruit of the Momordica charantia in type 2 diabetic mice[J].J Nutr Sci Vitaminol,2001,47(5):340-344
46 Chen QX,Chan LLY,Li-ETS,et al.Bitter melon(Momordica charantia) reduces adiposity,lowers serum insulin and normalizes glucose tolerance in rats fed a high fat diet[J].J Nutr,2003,133:1088-1093
47 Welihinda J,Karunanayake EH,Sheriff MH,et al.Effect of Momordica charantia on the glucose tolerance in maturity onset diabetes[J].J Ethnopharmacol,1986,17:277-282
48 Day C,Cartwright T,Provost J,et al.Hypoglycaemic effect of Momordica charantia extracts[J].Planta Medica,56:426-429
49 Cakici I,Hurmoglu C,Tunctan B,et al.Hypoglycaemic effect of Momordica charantia extracts in normoglycaemic or cyproheptadineinduced hyperglycaemic mice[J].J Ethnopharmacol,1994,44:117-121
50 Raza H,Ahmed I,John A,et al.Modulation of xenobiotic metabolism and oxidative stress in chronic streptozotocin-induced diabetic rats fed with Momordica charantia fruit extract[J].J Biochem Mol Toxicol,2000,14(3):131-139
51 范玉玲,崔福德.苦瓜有效部位降糖活性的比较研究[J].沈阳药科大学学报,2001,18(1):50-52
52 Vikrant V,Grover JK,Tandon N,et al.Treatment with extracts of Momordica charantia and Eugenia jambolana prevents hyperglycemia and hyperinsulinemia in fructose fed rats[J].J Ethnopharmacol,2001,76(2):139-143
53 Kar A,Choudhary BK,Bandyopadhyay NG.Comparative evaluation of hypoglycaemic activity of some Indian medicinal plants in alloxan diabetic rats[J].J Ethnopharmacol,2003,84:105-108
54 Khan B,Arayne MS,Naz S,et al.Hypogylcemic activity of aqueous extract of some indigenous plants[J].Pak J Pharm Sci,2005,18(1):62-64
55 Mahomoodally MF,Gurib-Fakim A,Subratty AH.Effect of exogenous ATP on Momordica charantia Linn.(Cucurbitaceae) induced inhibition of d-glucose,1-tyrosine and fluid transport across rat everted intestinal sacs in vitro[J].J Ethnopharmacol,2007,110(2):257-263
56 Abd El Sattar El Batran S,El-Gengaihi SE,El Shabrawy OA.Some toxicological studies of Momordica charantia L.on albino rats in normal and alloxan diabetic rats[J].J Ethnopharmacol,2006,108(2):236-242
57 Sarkar S,Pranava M,Marita R.Demonstration of the hypoglycemic action of Momordica charantia in validated animal model of diabetes[J].Pharmacol Res,1996,33(1):1-4
58 王先远,金宏.苦瓜皂甙降血糖作用及其机制初探[J].氨基酸和生物资源,2001,23(3):42-45
59 Higashino H,Suzuki A,Tanaka Y,et al.Hypoglycemic effects of Siamese Momordica charantia and Phyllanthus urinaria extracts in streptozotocin induced diabetic rats[J].Nippon Yakurigaku Zasshi,1992,100:415-421
60 Chaturvedi P,George S,Milinganyo M,et al.Effect of Momordica charantia on lipid profile and oral glucose tolerance in diabetic rats[J].Phytother Res,2004,18(11):954-956
61 Tongia A,Tongia S,Dave M,et al.Phytochemical determination and extraction of Momordica charantia fruit and its hypoglycemic potentiation of oral hypoglycemic drugs in diabetes mellitus(NIDDM)[J].Indian J Physiol Pharmacol,2004,48:241-244
62 Lotlikar MM,Rajarama Rao MR.Pharmacology of a hypoglycemic principle isolated from the fruits of Momordica charantia L[J].Indian J Pharm,1966,(28):129-132
63 Olaniyi AA.A neutral constituent of Momordica foetida[J].Lloydia,1975,38:361-362
64 Ng TB,Wong CM,Li WW,et al.A steryl glycoside fractions from Momordica charantia seeds with an inhibitory action on lipid metabolism in vitro[J].Biochem Cell Biol,1986,64(8):766-771
65 张萍萍,王峰,薛爱琴等.苦瓜素降血糖作用的试验研究[J].江苏中医,1997,7:30-31
66 孙宝莹,孙宝全.苦瓜皂甙的药效学实验研究[J].河南中医药学刊.1994,9(6):19-21
67 Matsuda H,Li Y,Murakami T,et al.Antidiabetic principles of natural medicines Ⅲ.Structure-related inhibitory activity and action model of oleanolic acid glycosides on hypoglycemic activity[J].Chem Pharm Bull,1998,46(9):1399-1403
68 赵海雯.苦瓜皂甙的分离纯化及降血糖功效研究[D].硕士学位论文,江南大学,2005
69 石雪萍.苦瓜皂甙的分离、纯化、结构鉴定及降血糖功能研究[D].博士学位论文,江南大学,2007
70 Baldwa VS,Goyal RK,Bhandari CM,et al.A clinical trial of insulin obtained from vegetable source (plant insulin) in patients with diabetes mellitus[J].Rajasthan Med J,1976,15(1):54-60
71 张世荣,姚文贞,薛庆富等.苦瓜降糖肽Ⅰ.分离纯化和特性[J].生物化学与生物物理学报,1980,12(4):291
72 王敬文,龚桂,陈随军等.植物降血糖活性蛋白新组分PA及其制备.专利申请号:93119351,1995-5
73 权建新,篙兴厚,张振强等.苦瓜果实中植物胰岛素降血糖研究.陕西医学杂志,1991,20(11):691-693
74 盛清凯,姚惠源.苦瓜中植物胰岛素的分离及其氨基酸序列测定[J].食品与发酵工业,2004,30(7):129-131
75 王勇庆.苦瓜降血糖作用研究[J].湖南中医杂志,1998,14:54-55
76 Zheng ZX,Teng JY,Liu JY,et al.The hypoglycemic effects of crude polysaccharides extract from Momordica charantia in mice[J].Wei Sheng Yan Jiu,2005,34(3):361-363
77 American Diabetes Association.Screening for type 2 diabetes.Diabetic Care,1998,21:S20-S31
78 World Health Organization,1994.Prevention of diabetes mellitus.Technical Report Series NO 844.Geneva,WHO
79 Grover JK,Rathi SS,Vats V.Amelioration of experimental diabetic neuropathy and gastropathy in rats following oral administration of plant(Eugenia Jambolana,Mucuna pruriens and Tinospora cordifolia)extracts[J].Indian Journal of Experimental biology,2002,40:273-276
80 Grover JK,Vats V,Rathi SS,et al.Traditional Indian anti-diabetic plants attenuate progression of renal damage in streptozotocin-induced diabetic mice[J].Journal of Ethnopharmacology,2001,76:233-238
81 Feldman M,Schiller ER.Disorders of gastrointestinal motility associated with diabetes mellitus[J].Annals of Internal Medicine,1983,98:911-922
82 Hiller R,Sperduto RD,Ederer F.Epidemiologic associations with cataract in the 1971-1972.National Health and Nutrition Examination Survey[J].American Journal of Epidemiology,1983,118:239-249
83 Van-Heyningen R,Harding JJ.A case-control study of cataract on Ofordshire:some risk factors[J].British Journal of Opthalmology,1988,72:804-808
84 Harding JJ,Harding RS,Egerton M.Risk factors for cataract in Oxfordshire:diabetes,peripheral neuropathy,myopia,glaucoma and diarrhea[J].Acta Ophthalmolgica(Copenhagen),1989,67:517-518
85 Rathi SS,Grover JK,Vikrant V.Prevention of experimental diabetic cataract by Indian Ayurvedic plant extracts[J].Phytotherapy Research,2002b,16:774-777
86 Srivastava Y,Venkatakrishna-Bhatt H,Verma Y.Effect of Momordica charantia Linn.pomous aqueous extract on cataractogenesis in murrin alloxan diabetics[J].Pharmacol Res Commun,1988,20(3): 201-209
87 Licastro F,Franceschi C,Barbieri L,et al.Toxicity of Momordica charantia lectin and inhibitor for human normal and leukaemic lymphocytes[J].Virchows Archives of B Cell Pathology Including Molecular Pathology,1980,33:257-265
88 Ng TB,Liu WK,Sze SF,et al.Action of alpha-momorcharin,a ribosome inactivating protein,on cultured tumor cell lines[J].General Pharmacology,1994,25:75-77
89 Battelli MG,Polito L,Bolognesi A,et al.Toxicity of ribosome-inactivating proteins- containing immunotixins to a human bladder carcinoma cell line[J].International Journal of Cancer,1996,68:485-490
90 Tsao SW,Ng TB,Yeung HW.Toxicities of trichosanthin and alpha-momorcharin,abortifacient proteins from Chinese medicinal plants,on cultured tumor cell lines[J].Toxicon,1990,28:1183-1192
91 Sun Y,Huang PL,Li JJ,et al.Anti-HIV agent MAP30 modulates the expression profile of viral and cellular genes for proliferation and apoptosis in AIDS-related lymphoma cells infected with Kaposi's sarcoma-associated virus[J].Biochemical and Biophysical Research Communication,2001,287:983-994
92 Ganguly C,De S,Das S.Prevention of carcinogen-induced mouse skin papilloma by whole fruit aqueous extract of Momordica charantia[J].European Journal of Cancer Prevention,2000,9:283-288
93 Nagasawa H,Watanabe K.Inatomi H.Effects of bitter melon(Momordica charantia L.) or ginger rhizome(Zingbier offifinale rocs) on spontaneous mammary tumorigenesis in SHN mice[J].American Journal of Chinese Medicine,30:195-205
94 Claflin AJ,Vesely DL,Hudson JL,et al.,Inhibition of growth and guanylate cyclase activity of an undifferentiated prostate adenocarcinoma by an extract of the balsam pear(Momordica charantia abbreviata)[J].Proc Natl Acad Sci USA,1978,75(2):989-993
95 严启新,罗天浩.苦瓜的研究及开发思考[J].时珍国药研究,1997,8(4):380-381
96 Basch E,Gabardi S,Ulbricht C.Bitter melon(Momordica charantia):a review of efficacy and safety[J].American Journal of Health and Systemic Pharmacology,2003,65:356-359
97 Takemoto DJ,Dunford C,McMurray MM.The cytotoxic and cytostatic effects of the bitter melon on human lymphocytes[J].Toxicon,1982a,20(3):593-599
98 Takemoto DJ,Dunford C,Vaughn D,et al.Guanylate cyclase activity in human leukemic and normal lymphocytes:Enzyme inhibition and cytotoxicity of plant extracts[J].Enzyme,1982,27:179-188
99 齐文波,徐中干,徐誉泰等.苦瓜素的分离纯化与抗肿瘤活性的研究[J].离子交换及吸附,1999,15(1):59-63
100 Lee Huang S,Huang PL,Chen HC,et al.Anti-HIV and anti-tumor activities of recombinant MAP30 from bitter melon[J].Gene,1995a,61(2):151-156
101 Lee Huang S,Huang PL,Bourinbaiar,AS,et al.Inhibition of the integrase of human immunodeficiency virus(HIV) type 1 by anti-HIV plant protion MAP30 and GAP31[J].Proc Natl Acad Sci USA,1995b,92(19):8818-8822
102 Huang PL,Sun Y,Chen HC,et al.Proteolytic fragments of anti-HIV and anti-tumor proteins MAP30 and GAP31 are biologically active[J].Biochem Biophys Res Commun,1999,262:615-623
103 陈执中.抗艾滋病天然药物-苦瓜成分的研究Ⅰ,水溶性成分的分析[J].中国民族民间医药杂志,1999,37:63-65
104 Bourinbaiar AS,Lee Huang S.The activity of plant-derived antiretroviral protens MAP30 and GAP31against herpes simplex virus in vitro[J].Biochem Biophys Res Commun,1996,219(3):923-929
105 Zheng YT,Ben KL,Jin SW.Alpha-momorcharin inhibits HIV-1 replication in acutely but not chronically infected T lymphoid cells[J].Acta Pharm Sin,1999,20:239-243
106 Au TK,Collins RA,Lain TL,et al.The plant ribosome inactivating proteins luffin and saporin are potent inhibition of HIV-1 integrase[J].FEBS Letters,2000,471:169-172
107 Bourinbaiar AS,Lee-Huang S.Potentiation of anti-HIV activity of anti-inflammatory drugs,dexamethasone and indomethacin,by MAP30,the antiviral agent from bitter melon[J].Biochemistry and Biophysics Research Communication,1995,208:779-785
108 Wang HX,Ng TB.Examination of lectins,polysac charopeptide,polysaccharide,alkaloid,coumarin and trypsin inhibitors for inhibitory activity against human immunodeficiency virus reverse transcriptase and glycohydrolases[J].Planta Medica,2001a,67:669-672
109 张素,王明霞.植物药材中抗艾滋病毒活性成分研究[J].中医药信息,1994,11(4):30-33
110 Foa TL,Campadelli FG,Barbieri L,et al.Effect of ribosome-inactivating proteins on virus-infected cells:inhibition of virus multiplication and protein synthesis[J].Arch Virol,1982,71:322-332
111 Khan MR,Omoloso AD.Momordica charantia and Allium sativum:Broad spectrum antibacterial activity [J].Korean J Pharmacog,1998,29(3):155-58
112 Ogata F,Miyata T,Fujii N,et al.Purification and amino acid sequence of a bitter gourd inhibitor against an acidic amino acid-specific endopeptidase of Streptomyces griseus[J].Journal of Biological Chemistry,1991,266:16715-16721
113 Omoregbe RE,Ikuebe OM,Ihimire IG.Antimicrobial activity of some medicinal plants extracts on Escherichia coli,Salmonella paratyphi and Shigella dyseteriae[J].African Journal of Medical Science,1996,25:373-375
114 Yesilada E,Gurbuz I,Shibata H.Screening of Turkish anti-ulcerogenic folk remedies for anti-Helicobacter pylori activity[J].J Ethnopharmacol,1999,66:289-293
115 吴传茂,夏服宝,李小燕.苦瓜的抑菌作用和苦瓜茶[J].中国商办工业,2002,3:51-52
116 张绪忠,陈青山.苦瓜抑菌防腐作用的实验研究[J].预防医学情报杂志,1996,12(1):17-20
117 黎锦城.苦瓜的抗炎作用[J].中药材,2001,24(7):508-509
118 何煜波.苦瓜抗菌作用研究[J].食品科学,1998,19(3):34-36
119 陈清山,程光文,张绪忠.不同苦瓜提取液对肉类食品防腐作用的研究[J].武汉冶金科技大学学报,1996,19(3):387-390
120 陈冀胜,郑硕.中国有毒植物[M].北京:科学出版社,1987,PP.128
121 覃国芳,李远慧,高晓勤等.苦瓜果对雄性大鼠生育力的影响[J].贵阳医学院学报,1995,20(1):14-17
122 Chan WY,Tam PP,Choi HL,et al.Effects of momorcharins on the mouse embryo at the early organogenesis stage[J].Contraception,34:537-544
123 王庆华,于长春,徐誉泰等.α-苦瓜素和β-苦瓜素的研究进展[J].中草药,1995,26(5):266-267
124 Law LK,Tam PP,Yeung HW.Effects of alpha-trichosanthin and alpha-momorcharin on the development of peri-implantation mouse embryos[J].J Reprod Fert,1983,69:597-604.
125 Tam PP,Law LK,Yeung HW.Effects of alpha-momorcharin on preimplantation development in the mouse[J].J Reprod Fert,1984,71:33-38
126 常凤岗,李建梅.苦瓜抗生育活性成分的化学研究(Ⅰ)[J].中草药,1995,21(6):281-284
127 Schreiber CA,Wan L,Sun Y,et al.The antiviral agents,MAP30 and GAP31,are no toxic to human spermatozoa and may be useful in preventing the sexual transmission of human immunodeficiency virus type 1[J].Fertil Steril,1999,72(4):686-690
128 乔昕,陈雄,李冬生.苦瓜抗氧化作用的初步研究[J].湖北农业科学,2003,1:74-75
129 傅明辉.苦瓜籽蛋白的分离、纯化及其抗氧化作用的研究[J].药物生物技术,2001,8(5):248-250
130 傅明辉,陈健汉,庄东红.苦瓜提取液的抗氧化、抑菌和降血糖活性[J].食品科学,2001,22(4):88-90
131 Leung SO,Yeung HW,Leung KN.The immunosuppressive activities of two abortifacient proteins isolated from the seeds of bitter melon(Momordica charantia)[J].Immunopharmacology,1987,13(3):159-171
132 Spreafico F,Malfiore C,Moras ML.The immunomodulatory activity of the plant proteins Momordica charantia inhibitor and pokeweed antiviral protein[J].International Journal of Immunopharmacology,1983,5:335-343
133 Cunnick JE,Sakamoto K,Chapes SK.Induction of tumor cytotoxic immune cells using a protein from the bitter melon(Momordica charantia)[J].Cellular Immunology,1990,126:278-289
134 程光文,陈清山,张绪忠.苦瓜对正常小鼠和免疫受抑小鼠免疫促进作用的实验观察[J].中国冶金工业医学杂志,1996,13(4):198-200
135 Sitasawad SL,Shewade Y,Bhonde R.Role of bittergourd fruit juice in stz-induced diabetic state in vivo and in vitro[J].J Ethnopharmacol,2000,73(1-2):71-79
136 Welihinda J,Arvidson G,Gylfe E,et al.The insulin-releasing activity of the tropical plant Momordica charantia[J].Acta Biot Mel Ger,1982,41(12):1229-1240
137 Shibib BA,Khan LA,Rahan R.Hypoglycaemic activity of Coccinia indica and Momordica charantia on diabetic rats:depression of the hepatic gluconeogenic enzymes glucose-6-phosphatase and fructose-1,6-biphosphatase and elevation of both liver and red-cell shunt enzyme glucose-6-phosphate dehydrogenase[J].Biochemistry Journal,1993,292:267-290
138 袁宝君,戴月.癞葡萄胶囊调节血糖人体试食试验的研究[J].卫生研究,2003,32:326-328
139 石根勇,吕中明,陈新霞等.癞葡萄冻干粉凋节血糖作用的研究[J].中国生化药物杂志,2005,26(5):303-304
140 冯怀蓉,张慧涛,茆军.多肽简介及应用[J].新疆农业科学,2002,39(1):38-39
141 冯秀燕,计成.寡肽在蛋白质营养中的作用[J].动物一样学报,2001,13(3):8-13
142 Webb KE Jr,DiRienzo DB,Mathews JC.Recent development in gastrointestinal absorption and tissue utilization of peptides:a review[J].J Dairy Sci,1993,76:351-361
143 Gobbetti M,Stepaniak L,de Angelis M.Latent bioactive peptides in milk proteins:proteolytic activation and significance in dairy processing[J].Cri Rev Food Sci Nutri,2002,42(3):223-239
144 Vincenzini MT,Iantomasi T,Favilli F.Glutathione transport across intestinal brush-border membranes:effects of ions,pH,delta psi,and inhibitors[J].Biochim Biophys Acta,1989,987(1):29-37
145 Dantzing et al.Association of intestinal peptide transport with a protein related to the cadheririn super family[J].Science,1994,764:430-433
146 蒲首丞,王金水.生物活性肽的制备与应用[J],2005,5:49-51
147 刘平,田桂玲,叶蕴华.固载化酶催化合成多肽的研究进展[J].高等学校化学学报,2001,8:1342-1348
148 Meisel H.Biochemical properties of regulator peptides derived from milk proteins[J].Biopolymers,1997,43(2):119-125
149 Meisel H,Bockelmann W.Bioactive peptides encrypted in milk proteins proteolytic activation and thropho-functional properties[J].Antonie Van Leeuwenhoek,1999,76:207-213
150 Gill I,Lopez-Fandino R,Jorba X.Biologically active peptides and enzymatic approaches to their production[J].Enzyme and Microbial Technology,1996,18:162-183
151 Korhonen H,Pihlanto A.Food-derived bioactive peptides-opportunities for designing future foods[J].Current Pharmaceutical Design,2003,9:1297-1308
152 邵丙扬.警惕世界面临糖尿病灾难[J].内科急危重症杂志,2004,10(1):1-2
153 Zhang BB,Moller DE.News approaches in the treatment of type 2 diabetes[J].Curr Opin Chem Biol,2000,4:461-467
154 Tolman KG,Chandramouli J.Hepatotoxicity of the thiazolidinediones[J].Clin Liver Dis,2003,7:369-370.
155 王吉影,顾耀,徐乃馨等.阿卡波糖和伏格列波糖治疗2型糖尿病的比较[J].同济大学学报(医学版),2001,2(21):32-35
156 Delea TE,Edelsberg JS,Hagiwara M,et al.Use of thiazolidinediones and risk of heart failure in people with type 2 diabetes:a retrospective cohort study[J].Diabetes Care,2003,26:2983-2989
157 张冬英.普洱茶降糖降脂活性成分的研究[D].博士学位论文,湖南农业大学,2006
158 Yoshikawa M,Fujita H,Matoba N,et al.Bioactive peptides derived from food protein preventing lifestyle-related diseases[J].Biofactors,2000,12:143-146.
159 Dziuba J,Minkiewicz P,Nalecz D.Biologically active peptides derived from plant and animal proteins [J].Pol Food Nutri Sci,1999,8:3-16
1 范冠杰主编.糖尿病[M].北京:人民卫生出版社,2002
2 Piedrola G,Novo E,Escober F,et al.White blood cell count and insulin resistance in patients with coronary artery disease[J].Annual Endocrinology(Paris),2001,62:7-10
3 Yaryur-Tobias JA,Pinto A,Neziroglu F.Anorexia nervosa,diabetes mellitus,brain atrophy and fatty liver [J].International of Etiological Disorders,2001,30:350-353
4 Akhtar MS,Iqbal J.Evaluation of the hypoglycemic effect of Achyranthes aspera in normal and alloxan diabetic rabbits[J].J Ethnopharmacol,1991,31:49-57
5 Tolman KG,Chandramouli J.Hepatotoxicity of the thiazolidinediones[J].Clinics in Liver Disease,2003,7:369-370
6 Zhang BB,Moller DE.News approaches in the treatment of type 2 diabetes[J].Curr Opin Chem Biol,2000,4:461-467
7 Li WL,Zheng HC,Bukuru J,et al.Natural medicines used in the traditional Chinese medical system for therapy of diabetes mellitus[J].J Ethnopharmacol,2004,92:1-21
8 Ugochukwu NH,Babady NE.Antihyperglycemic effect of aqueous and ethanolic extracts of Gongronema latifolium leaves on glucose and glycogen metabolism in livers of normal and streptozotocin-induced diabetic mice[J].Life Sci,2003,73:1925-1938
9 Eddouks M,Lemhadri A,Zeggwagh NA,et al.Potent hypoglycaemic activity of the aqueous extract of Chamaemelum nobile in normal and streptozotocin-induced diabetic rats[J].Diabetes Res Clin Pract,2005,67(3):189-195
10 Zhang RX,Zhou JH,Jia ZP,et al.Hypoglycemic effect of Rehmannia glutinosa oligosaccharide in hyperglycemic and alloxan-induced diabetic rats and its mechanism[J].J Ethnopharmacol,2004,90:39-43
11 Sezik E,Aslan M,Yesilada E,et al.Hypoglycaemic activity of Gentiana olivieri and isolation of the active constituent through bioassay-directed fractionation techniques[J].Life Sci,2005,76:1223-1238
12 Lee MK,Kim MJ,Cho SY,et al.Hypoglycemic effect of Du-zhong(Eucommia ulmoides Oliv.) leaves in streptozotocin-induced diabetic rats[J].Diabetes Res Clin Pract,2005,67:22-28
13 Roy S,Sehgal R,Padhy BM,et al.Antioxidant and protective effect of latex of Calotropis procera against alloxan-induced diabetes in rats[J].J Ethnopharmacol,2005,102:470-473
14 Joy KL,Kuttan R.Anti-diabetic activity of Picrorrhiza Kurroa extract[J].J Ethnopharmacol,1999,167:143-148
15 Yadav S,Vats V,Dhunnoo Y,et al.Hypoglycemic and antihyperglycemic activity of Murraya koenigii leaves in diabetic rats[J].J Ethnopharmacol,2002,82:111-116
16 Ali L,Khan AK,Mamun MI,et al.Studies on hypoglycemicin effects of fruit pulp seed and whole plant of Momordia charantia on normal and diabetic model rats[J].Planta Med,1993,59:408-412
17 Jouad H,Eddouks M,Lacaille-Dubois MA.Hypoglycaemic effect of the water extract of Spergularia purpurea in normal and streptozotocin-induced diabetic rats[J].J Ethnopharmacol,2000,71:169-177
18 李聪然,游雪甫,蒋建东.糖尿病动物模型及研究进展[J].中国比较医学杂志,2005:15(1):59-63
19 黄伟坤.食品检验与分析[M].北京:中国轻工业出版社,1997
20 赵亚华,高向阳.生物化学实验技术教程[M].广州:华南理工大学出版社,2000
21 郑建仙.功能性食品[M].北京:中国轻工业出版社,1999,486-487
22 吴有炜.试验设计与数据处理[M].苏州:苏州大学出版社,2002
23 陈晓超.苦瓜提取物中降糖物质的分离鉴定及其降糖机理的初步探讨[D].福州大学,硕士学位论文,2004
24 http://sun.ars-grin.gov:8080/npgspub/xsql/duke/plantdisp.xsql?taxon=694
25 http://sun.ars-grin.gov:8080/npgspub/xsql/duke/plantdisp.xsql?taxon=843
26 Takasu N,Komiya I,Asawa T,et al.Streptozocin and alloxan-induced H_2O_2 generation and DNA fragmentation in pancreatic islets[J].Diabetes,1991,40(9):1141-1145
27 潘竞锵,韩超,刘惠纯等.菌陈蒿汤对正常和多种糖尿病模型动物血糖的影响[J].中药材,2001, 24(2):128-131
28 蒋渝,周永禄.四氧嘧啶诱发糖尿病模型的建立[J].上海实验动物科学,]990,10(2):79-81
29 张传仓,刘卫鹏,杜江等.改良四氧嘧啶法制作糖尿病动物模型[J].实用医学杂志,2007,23(5):623-624
30 Utkan T,Yildirim MK,Yildirim S,et al.Effects of the specific phosphodiesterase inhibitors on alloxan-induced diabetic rabbit cavernous tissue in vitro[J].Int J Impot Res,2001,13(1):24-30
31 Vats V,Grover JK,Rathi SS.Evaluation of anti-hyperglycemic and hypoglycemic effect of Trigonella foenum-graecum Linn,Ocimum Sanctum Linn and Pterocarpus marsupium Linn,in normal and alloxanized diabetic rats[J].J Ethnopharmacol,2002,79:95-100.
32 金丕焕.医用统计方法(第二版)[M].上海:复旦大学出版社,2003
33 冯礼明.单一自由度比较分析方法介纠[J].数理统计与管理,1990,1:21-26
34 周乃健,赵志立.单一自由度分组比较统计分析方法在农业试验中的应用[J].山西农业大学学报(自然科学版),1992,12(2):110-114
35 Sitasaward SL,Shewade Y,Bhonde R.Role of bittergourd fruit juice in STZ-induced diabetic state in vivo and in vitro[J].J Ethnopharmacol,2000,73:71-79
36 Chattopadhyay RR,Sarkar SK,Ganguly S,et al.Hypoglycemic and antidiabetic effects of leaves of Vinca rosea Linn[J].Indian Journal of Physiology and Pharmacology,1991,35:145-151
37 Sharma SR,Dwivedi SK,Swarup D.Hypoglycaemic and hypolipidemic effects of Cinnamomum tamala Nees leaves[J].Indian Journal of Experimental Biology,1996,34:372-374
38 Rao VV,Dwivedi SK,Swarup D,et al.Hypoglycemic and antidiabetic effects of Aegle marmelose leaves in rabbits[J].Current Science,1995,69:932-933
39 张芳林,李果,刘优萍等.2型糖尿病大鼠模型的建立及其糖代谢特征分析[J].中国实验动物学报,2002,10(1):16-20
40 池芝盛主编.糖尿病学(第1版)[M].北京:人民卫生出版社,1986:161-162
1 汪家政,范明主编.蛋白质技术手册[M].北京:科学技术出版社,2000
2 赵锐,顾谦群,管华师.多肽物质分离与分析方法研究进展[J].中国海洋药物,2000,3:48-53
3 Nebot VS,Benavente F,Hernadex E,et al.Evaluation of the electrophoretic behaviour of opioid peptides separation by capillary electrophoresis-electrospray ionzation mass spectrometry[J].Analytical Chimmica Acta,2006,577:68-76
4 Herraiz T.Sample preparation and reversed phase-high performance liquid chromatography analysis of food-derived peptides[J].Analytica Chimica Acta,1997,352:119-139
5 Bayerbach R,Nguyen VD,Schurr U,et al.Characterization of the water-insoluble fraction from fast pyrolysis liquids(pyrolytic lignin) part Ⅲ.Molar mass characteristics by SEC,MALDI-TOF-MS,LDI-TOF-MS,and Py-FIMS.J Anal Appl Pyrolysis,2006,77(2):95-101
6 Sporns P,Wang J.Exploring new frontiers in food analysis using MALDI-MS[J].Food Res Int,1998,31(3):181-189
7 时钧、袁权.膜技术分离手册[M].北京:化学工业出版社,2001
8 刘茉娥.膜分离技术[M].北京:化学工业出版社会,1998
9 Rautenback R,Albrecht R.Membrane Process[M].Chichester:John Wiley and Sons Ltd,1989,pp.2-15
10 Kawakatsu T,Kobayashi T,Sano Y,et al.Clarification of green tea extract by microfiltration and ultrafiltration[J].Biosci Biotech Biochem,1995,59(6):1016-1020
11 张晓楠,郭立安.超滤在蛋白质纯化中的应用[J].中国生化药物杂志,1999,20(2):97-98
12 Alvise ND,Lesueur-Lambert C,Fertin B.Hydrolysis and large scale ultrafiltration study of alfalfa protein concentrate enzymatic hydrolysate[J].Enzyme and Microbial Tehcnology,2002,27:286-294
13 刘忠洲,续曙光,李锁定.微滤、超滤过程中的膜污染与清洗[J].水处理技术,1997,23(4):187-193
14 郑成.膜的清洗及杀菌[J].化工进展,1996,6:45-49
15 邬行彦,孙进.膜过滤法及其在抗生素工业中的应用[J].国外医药抗生素杂志分册,1992,13(6):455-462;1993,14(1):60-67
16 刘茉娥.超滤膜污染机理的研究与控制[J].水处理技术,1989,3:163-169
17 张赞红,施亚钧.超滤膜分离技术的应用及膜污染问题[J].化学世界,1993,6:242-245
18 任建新.膜分离技术及其应用[M].北京:化学工业出版社,2003:1
19 华耀祖.超滤技术与应用[M].北京:化学工业出版社工业装备与信息工程出版中心,2004
20 Rao A,Shallo HE,Ericson HE,et al.Characterization of soy protein concentrate produced by membrane ultrafiltration[J].Journal of Food Science,2002,67(4):1412-1418
21 Shallo HE,Rao A,Ericson AP,et al.Preparation of soy protein concentrate by ultrafiltration[J].Journal of Food Science,2001,66(2):242-246
22 龙晓丽.红三鱼蛋白水解物的制备及分离技术的研究[D].硕士学位论文,华南理工大学,2004
23 江华.超滤及冷冻浓缩技术在低聚木糖制备中的应用[D].博士学位论文,南京林业大学,2001
24 赵永芳.生物化学技术原理及其应用[M].武汉大学出版社,1997
25 Irvine GB.High-performance size-exclusion chromatography of peptides[J].J Biochem Biophys Methods,2003,56:233-242
26 韩冬,叶美玲,施良和.水溶性凝胶色谱中的非体积排阻效应[J].色谱,1995,13(6):432-436
27 Dan N,Ganesan R,Flood KG,et al.Determination of enantiomers in a synthetic argininal peptide using capillary zone electrophoresis and high-performance liguid chromatography[J].J Chromatogr,2000,891:115-120
28 Sanz NV,Benavente F,Toro I,et al.Optimization of HPLC conditions for the separation of complex crude mixtures produced in the synthesis of therapeutic peptide hormones[J].Chromatogr,2000,891:115-120
29 Smith DD,Saha S,Fang G,et al.Modifications to the N-terminus but not the C-terminus of calcitonin gene-related peptide(8-37) produce antagonists with increased affinity[J].J Med Chem,2003,46:2427-2435
30 Purcell AW,Zhao GL,Aguilar MI,et al.Comparison between the isocratic and gradient retention behavior of polypeptides in reversed-phase liquid chromatographic environments[J].J Chromatogr,1999,852:43-46
31 王英武,王玲,顾景凯等.电喷雾-串联四极杆-飞行时间质谱法分析寡肽的一级结构[J].分析化学,2003,31(6):709-712
32 邓慧敏,赖志辉,黎军等.碎片结构分析在MALDI-TOF-MS法测定多肽序列中的应用[J].生物化学与生物物理学报,2000,32(2):179-182
33 Medzihradszky KF,Adems GW,Burlingame AL,et al.Peptide sequence determination by matrix-assisted laser desorption ionization employing a tandem double focusing magnetic orthogonal acceleration time-of-flight mass spectrometry[J].J Am Soc Mass Spetrom,1996,7:1-10
34 Kaufmann R.Maxtrix-assisted laser desorption ionization(MALDI) mass spectrometry:a novel analytical tool in molecular biology and biotechnology[J].J Biotechnol,1995,41:155-175
35 卞则梁,王光辉.质谱与生命科学[J].化学通服,1994,7:40-46
36 桑志红,杨松成.电喷雾电离质谱及其在质白质化学研究中的应用[J].国外医学药学分册,2000,27(1):38-41
37 梁宋平.世纪之交的蛋白质序列测定技术[J].生命科学,1999,1:31-34
1 Yoshikawa M,Fujita H,Matoba N,et al.Bioactive peptides derived from food protein preventing lifestyle-related diseases[J].Biofactors,2000,12:143-146.
2 Dziuba J,Minkiewicz P,Nalecz D.Biologically active peptides derived from plant and animal proteins [J].Pol Food Nutri Sci,1999,8:3-16
3 Suetsuna K,Chen JR.Isolation and characterization of peptides with antioxidant activity bioprocesses used in isolation and recovery[J].Curr Pharm Des,2003,9(16):1309-1323
4 Mahmoud MI.Physicochemical and functional properties of protein hydrolysates in nutritional products [J].Food Technol,1994,48:89-95
5 夏书华.螺蛳中生物活性物质的研究[D].博士学位论文,江南大学,2006
6 孔祥珍.小麦面筋蛋白的酶解及其阿片肽的研究[D].博士学位论文,江南大学,2006
7 Suetsuna K,Chen JR.Isolation and characterization of peptides with antioxidant activity derived from wheat gluten[J].Food Science and Technology Research,2002,8(3):227-230
8 Schusdziarra V,Schick R,Holand A,et al.Effect of opiate-active substances on pancreatic polypeptide levels in dogs[J].Peptides,1983,4:205-210
9 Nissen JA.Enzymatic hydrolysis of food protein.New York:Elsevier Applied Science Pulisher,1986
10 黄伟坤.食品检验与分析[M].北京:中国轻工业出版社,1990
11 李朝兴,印寿根,王克雷等.大孔吸附树脂对绞股蓝皂甙的吸附研究[J].离子交换与吸附,1994,10(3):203-207
12 赵亚华.生物化学实验技术教程[M].广州:华南理工大学出版社,2000
13 王章存,姚惠源.大米蛋白质的酶法水解及其性质研究[J].中国粮油学报,2003,18(5):5-7
14 程云辉.麦胚蛋白酶解物的制备、结构及其生物活性功能的研究[D].博士学位论文,江南大学,2006
15 Marquez MC,Vazquez MA.Modeling of enzymatic protein hydrolysis[J].Process Biochem,1999,35:111-117
16齐崴,何志敏,何丽霞.酪蛋白酶解制备酪蛋白磷酸肽研究Ⅰ过程分析与优化[J].食品科学,2001,22(7):25-28
17 Godfrey T,Reichelt J.Industrial Enzymology:the application of enzymes in industry.1983.England,Macmillan Publishers Ltd.pp 539-540,497-498
18王璋编.食品酶学[M].北京,轻工业出版社,1990,pp 180-185
19 Adamson NJ,Reynolds EC.Characterization of casein phosphopeptides prepared using Alcalase:determination of enzyme specificity[J].Enzyme and microbial technology,1996,19:202-207
20 Triveni R,Shamala TR,Rastogi NK.Optimised production and utilization of exopolysaccharide from Agrobacterium radiobacter[J].Process Biochemistry,2001,36:787-795
21 Kilic M,Bayraktar E,Ates S.Investigation of extractive citric acid fermentation using response-surface methodology[J].Process Biochemistry,2002,37:759-767
22 Mizubuti IY,Biondo Juniora O,de Oliveira Souza LW,et al.Response surface methodology for extraction optimization of pigeon pea protein[J].Food Chemistry,2000,70:259-265
23 Nag B,Medicherla S,Sharma SD.Orally active fraction of Momordica charantia,active peptides thereof,and their use in the treatment of diabetes[P].U.S.(P) US6127338,2000-10-03
24 Khanna P,Jain SC.Panangariya A.Hyopglycemic activity of polypeotide-P from a plant source[J].Nat Prod,1981,44:648-655
25 Owen R.Fennema著,王璋等译.食品化学(第三版)[M].北京:中国轻工业出版社,2003
26 邓劲光,胡熙恩,朱永.大孔吸附树脂对氢化可的松的吸附与洗脱性能研究[J].离子交换与吸附,2000,16(2):134-139
1 Charpentier G.Oral combination therapy from type 2 diabetes[J].Diabetes/Metabolism Research and Reviews,2002,18(Suppl 3):$70-$76
2 Ahmed I,Adeghate E,Sharma AK,et al.Effects of Momordica charantia fruit juice on islet morphology in the pancreas of the streptozotocin-diabetic rat[J].Diabetes Research Clinical Practices,1998,40:145-151
3 Welihinda J,Arvidson G,Gylfe E,et al.The insulin-releasing activity of the tropical plant Momordica charantia[J].Acta Biot Mel Ger,1982,41(12):1229-1240
4 Javill-Taylor KJ,Anderson RA,Graves DJ.A hydroxychalcome derived from cinnamon functions as a mimetic for insulin in 3T3-L1 adipocytes[J].Journal of the American College of Nutrion,2001,20:327-336
5 Qin B,Nagasaki M,Ren M,et al.Cinnamon extract(traditional herb) potentiates in vivo insulin-regulated glucose utilization via enhancing insulin signaling in rat[J].Diabetes Research and Clinical Practice,2003,63:139-148
6 Zhang RX,Zhou JH,Jia,ZP,et al.Hypoglycemic effect of Rehmannia glutinosa oligosaccharide in hyperglycemic and alloxan-induced diabetic rats and its mechanism[J].Journal of Ethnopharmacology,2004,90:39-43
7 沈同,王镜岩.生物化学[M].北京:高等教育出版社,1999
8 郑建仙.功能性食品[M].北京:中国轻工业出版社,1999,pp.486-487
9 Gold AH.The effect of diabetes and insulin on liver glycogen synthetase activation[J].Journal of Biological Chemistry,1970,245:903-905
10 Goel RK,Mahajan MP,Kulkarni SK.Evaluation of anti-hyperglycemic activity of some novel monocyclic beta lactams[J].J Pharm Pharm Sci,2004,7(1):80-83
11 Lolitkar MM,Rao MRR.Pharmacology of hypoglycaemic principle isolated from the fruits of Charantia Linn[J].Indian Journal Pharmacy,1966,28(5):129-133
12 Sharma SB,Nasir A,Prabhu KM,et al.Hypoglycaemic and hypolipidemic effect of ethanolic extract of seeds of Eugenia jambolana in alloxan-induced diabetic rabbits[J].Journal of Ethnopharmacology,2003,85:201-206
13关子安,孙茂欣,关大顺等.现代糖尿病学[M].天津:天津科学技术出版社,2000
14黄燕飞,刘志红.脂毒性与2型糖尿病及其并发症的关系[J].肾脏病与透析肾移植杂志,2002,11(1):58-63
15 Ginsberg HN.Basic mechanisms underlying the common hypertriglyceridemia and low HDL cholesterol levels[J].Diabetes,1996,45:S27-S30
16刘静,吴兆苏,赵冬.白由脂肪酸与代谢综合症的关系[J].心肺血管杂志,2005,24:125-127
17 Dhandapani S,Subramanian VR,Rajagopal S,et al.Hypolipidemic effect of Cuminum Cyminum L.on alloxan-induced diabetic rats[J].Pharmacological Research,2002,46(3):251-255
18 Bergman RN,Ader M.Free fatty acids and pathogenesis of type 2 diabetes mellitus[J].Trends Endocrinol Metab.2000.11:351-356
19邹效漫.二型糖尿病的脂蛋白代谢特点[A].见:叶平.血脂的基础与临床[M].北京:人民军医出版社,2002,191-200
20朱姿英,薛耀明.高甘油三酯血症和2型糖尿病[J].中国糖尿病杂志,2003,11(2):153-154
21 许樟荣,王爱红.糖尿病与血脂异常[J].中华老年医学杂志,2006,25(4)
22 Bopanna KN,Kannan J,Gadgil S,et al.Antidiabetic and antihyperglycemic effects of neem seed kernel powder on alloxan diabetic rabbits[J].Indian Journal of Pharmacology 1997,29:162-167
23 Luc G,Fruchart JC.Oxidation of lipoproteins and atherosclerosis.American Journal of Clinical Nutrition,1991,53:2065-2095
24 Ram A,Lauria P,Gupta R,et al.Hypocholesterolemic effects of Teminalia arjuna tree bark[J].Journal of Ethnopharmacology,1997,55:165-169
25 Sharma SR,Dwivedi SK,Swarup D.Hypoglycaemic,anti-hyperglycaemic and hypolipidemic activities of Caesalpinia sylvestre seeds in rats[J].Journal of Ethnopharmacology,1997,58:39-44
26 Jouad H,Lemhadri A,Maghrani M,et al.Cholesterol lowering activity of the aqueous extract of Spergularia in normal and recent onset diabetic rats[J].Journal of Ethnopharmacology,2003,87:43-49
27 Oberley LW.Free radicals and diabetes[J].Free Radical Biol and Med,1988,5:13 - 24
28 Jemings PE.The relationship of oxidative stress to thrombotic in type Ⅰ diabetic patients with diabetic retinopathy[J].Diabetic Med,1992,87:860 - 865
29 Maxwell SRJ,Thomason S,Sandier D,et al.Antioxidant status in patients with uncomplicated insulin-dependent and non-insulin-dependent diabetes mellitus[J].European Journal of Clinical Investigation,1997,27:484-490
30 Zhang XF,Tan BKH.Antihyperglycemic and antioxidant properties of Andrographis paniculata in normal and diabetic rats[J].Clinical and Experimental Pharmacology and Physiology,2000,27:358-363
31 El-Missiry MA,Othman AI,Amer MA.L-Arginine ameliorates oxidative stress in ailoxan-induced experimental diabetes mellitus[J].Journal of Applied Toxicology,2004,24:93-97
32 Manonmani G,Bhavapriya V,Kalpana S,et al.Antioxidant activity of Cassia Fistula(linn.) flowers in alloxan induced diabetic rats[J].Journal of Ethnopharmacology,2005,97:39-42
33 金晖,唐尧.糖尿病中氧化物与抗氧化物损伤[J].医学综述,1996,2(2):49-51
34 Vassalle C,Lubrano V,Abbate A,et al.Determination of metric plus nitrate and malondialdehyde in human plasma:analytical performance and the effect of smoking and exercise[J].Clin Chem Lab Med,2002,22(2):130-134
35 陈瑗.自由基医学基础与病理生理(第一版)[M].北京:人民卫生出版社,2002
36 丁爱玲,杨鄂平,吴丽霞等.中药对运动大鼠肾脏和肌肉自由基代谢的影响[J].北京体育大学学报,2004,27(2):206-208
37 刘方,武子斌,牛淑敏等.中药材抗氧化及自由基清除活性的研究[M].中国药学杂志,2001,36(7):442-445
38 Park SY,Bok SH,Jeon S,et al.Effect ofrutin and tannic acid supplements on cholesterol metabolism in rats[J].Nutrition Research,2002,22:283-295
39 Ohkawa H,Ohishi N,Yake K.Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction [J].Anal Biochem,1979,95:351-358
40 Chugh SN,Dhawan R,Kishore K,et al.Glibenclamide vs.gliclazide in reducing oxidative stress in patients of noninsulin dependent diabetes mellitus - a double blind randomized study[J].Journal of Association of Physicians of India,2001,49:803-807
41 Elmali E,Altan N,Bukan N.Effect of the sulphonylurea glibenclamide on liver and kidney antioxidant enzymes in streptozocin-induced diabetic rats[J].Drugs in R & D,2004,5(4):203-208
42 Asayama K,Nfeler F,Eenglish D,et al.Alloxan-induced free radical production in isolated cells,selective effect on islets cells[J].Diabetes,1984,33(10):1008-1011
43 蔡英兰,金政,金香子,等.榛花对糖尿病小鼠降糖作用及对肝脏和胰岛病理学影响[J].辽宁中医杂志,2002,32(12):1325-1326