柠檬酸对高脂血症小鼠胰岛素敏感性的影响
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摘要
目的:研究柠檬酸(citric acid,CA)调节血脂的作用和对高脂血症小鼠胰岛素敏感性的影响。方法:选用KM小鼠,建立高脂血症模型,以血脂康为阳性对照组,以低、中、高剂量柠檬酸干预模型小鼠,利用试剂盒测定血脂和胰岛素水平,并利用反转录聚合酶链式反应法测定小鼠肝脏6-磷酸葡萄糖酶(glucose-6-phosphatase,G-6-Pase)和四肢骨骼肌葡萄糖转运体4(glucose transporter-4,GLUT-4)mRNA的表达水平,考察柠檬酸对高脂血症小鼠血脂的影响以及对极胰岛素敏感性的改善作用。结果:1)与空白对照组相比,高脂模型组(hyperlipidemia model,HM)血清甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC)和低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)浓度极显著升高(P<0.01),高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)浓度极显著降低(P<0.01);与HM组相比,柠檬酸干预组(CA+HM)和阳性对照组血清TG、TC和LDL-C浓度显著降低(P<0.05),HDL-C的浓度极显著升高(P<0.01),但与空白对照组相比无显著差异(P>0.05);2)HM组空腹血糖浓度和灌糖2 h后的血糖浓度与空白对照组、阳性对照组以及CA+HM组相比极显著升高(P<0.01),且HM组对糖的代谢作用相对于其他组极显著减弱(P<0.01);3)与空白对照组相比,HM组空腹胰岛素(fasting insulin,FIN)含量和胰岛素抵抗指数(homeostatic model assessment of insulin resistance,HOMA-IR)极显著升高(P<0.01),胰岛素敏感指数(insulin sensitive index,ISI)极显著降低(P<0.01);与HM组相比,CA+HM组和阳性对照组的FIN含量、ISI和HOMA-IR有不同程度的升高或降低,其中CA+HM(H)组ISI极显著升高(P<0.01),FIN含量极显著降低(P<0.01),HOMA-IR显著降低(P<0.05),CA+HM(M)组ISI显著升高(P<0.05),FIN含量和HOMA-IR显著降低(P<0.05),CA+HM(L)组ISI显著升高(P<0.05),FIN含量极显著降低(P<0.01),HOMA-IR显著降低(P<0.05),但与空白对照组相比无显著差异(P>0.05);4)与空白对照组相比,HM组肝脏G-6-Pase mRNA表达量极显著增加(P<0.01);与HM组相比,CA+HM组和阳性对照组肝脏G-6-Pase mRNA表达量显著下调(P<0.05);与空白对照组相比,HM组四肢骨骼肌GLUT-4 mRNA表达量极显著降低(P<0.01);与HM组相比,CA+HM组和阳性对照组四肢骨骼肌GLUT-4 mRNA表达量显著上调(P<0.05)。结论:柠檬酸能够调节高脂血症小鼠的血脂水平,并能改善高脂血症小鼠对胰岛素的敏感性。
Objective: To explore the effect of citric acid(CA) on the regulation of blood lipids and insulin sensitivity in hyperlipidemic mice. Methods: KM mice were used to establish a hyperlipidemic animal model. Xuezhikang, a lipidlower drug in China, was used as a positive control, and low, medium and high-dose citric acid was given orally to the mouse model. Blood lipids and insulin levels were measured by commercial kits. The mRNA expression levels of glucose-6-phosphatase(G-6-Pase) in liver and glucose transporter-4(GLUT-4) in skeletal muscle were measured by reverse transcription-polymerase chain reaction(RT-PCR) to evaluate the effect of citric acid on blood lipids and insulin sensitivity in hyperlipidemic mice. Results: Compared with the blank control, the serum levels of triglyceride(TG), total cholesterol(TC) and low density lipoprotein cholesterol(LDL-C) in the hyperlipidemic model(HM) group was significantly increased,and serum HDL-C was significantly reduced(P < 0.01). Contrary results were observed when comparing the citric acid intervention and positive control groups with the HM group(P < 0.01), while there was no significant difference between the former ones and the blank control(P > 0.05). Compared with all other groups, fasting blood glucose(FBG) and 2 h postoral glucose tolerance test glucose levels were significantly higher in the HM group(P < 0.01), and glucose metabolism was significantly lower(P < 0.01). Fasting insulin(FIN) and homeostatic model assessment of insulin resistance(HOMA-IR) in the HM group were significantly higher and insulin sensitivity index(ISI) was significantly lower than in the blank control(P < 0.01). FIN, HOMA-IR and ISI in the citric acid intervention and positive control groups were different from those in the HM group; citric acid significantly increased ISI and decreased FIN and HOMA-IR(P < 0.01, P < 0.01 and P < 0.05 at high dose; P < 0.05, P < 0.05 and P < 0.05 at medium dose; P < 0.05, P < 0.01 and P < 0.05 at low dose) compared with the HM group, whereas all three dose groups showed no significant difference from the blank control(P > 0.05). The expression of G-6-Pase mRNA in liver in the HM group was significantly higher than in the blank control group(P < 0.01) and the citric acid intervention and positive control groups(P < 0.05), whereas the opposite was observed for the expression of GLUT-4 mRNA in skeletal muscle in the HM group versus the other groups. Conclusion: Citric acid can regulate blood lipid levels and improve insulin sensitivity in hyperlipidemic mice.
Objective: To explore the effect of citric acid(CA) on the regulation of blood lipids and insulin sensitivity in hyperlipidemic mice. Methods: KM mice were used to establish a hyperlipidemic animal model. Xuezhikang, a lipidlower drug in China, was used as a positive control, and low, medium and high-dose citric acid was given orally to the mouse model. Blood lipids and insulin levels were measured by commercial kits. The mRNA expression levels of glucose-6-phosphatase(G-6-Pase) in liver and glucose transporter-4(GLUT-4) in skeletal muscle were measured by reverse transcription-polymerase chain reaction(RT-PCR) to evaluate the effect of citric acid on blood lipids and insulin sensitivity in hyperlipidemic mice. Results: Compared with the blank control, the serum levels of triglyceride(TG), total cholesterol(TC) and low density lipoprotein cholesterol(LDL-C) in the hyperlipidemic model(HM) group was significantly increased,and serum HDL-C was significantly reduced(P < 0.01). Contrary results were observed when comparing the citric acid intervention and positive control groups with the HM group(P < 0.01), while there was no significant difference between the former ones and the blank control(P > 0.05). Compared with all other groups, fasting blood glucose(FBG) and 2 h postoral glucose tolerance test glucose levels were significantly higher in the HM group(P < 0.01), and glucose metabolism was significantly lower(P < 0.01). Fasting insulin(FIN) and homeostatic model assessment of insulin resistance(HOMA-IR) in the HM group were significantly higher and insulin sensitivity index(ISI) was significantly lower than in the blank control(P < 0.01). FIN, HOMA-IR and ISI in the citric acid intervention and positive control groups were different from those in the HM group; citric acid significantly increased ISI and decreased FIN and HOMA-IR(P < 0.01, P < 0.01 and P < 0.05 at high dose; P < 0.05, P < 0.05 and P < 0.05 at medium dose; P < 0.05, P < 0.01 and P < 0.05 at low dose) compared with the HM group, whereas all three dose groups showed no significant difference from the blank control(P > 0.05). The expression of G-6-Pase mRNA in liver in the HM group was significantly higher than in the blank control group(P < 0.01) and the citric acid intervention and positive control groups(P < 0.05), whereas the opposite was observed for the expression of GLUT-4 mRNA in skeletal muscle in the HM group versus the other groups. Conclusion: Citric acid can regulate blood lipid levels and improve insulin sensitivity in hyperlipidemic mice.
引文
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[20]郑铁生,王亚娜,宗爱萍.龙虾壳聚糖干预后糖尿病小鼠血糖和糖耐量的变化[J].中国临床康复,2006,10(31):67-69.
[21]李晓月,张晶晶,张红建,等.菊粉对反式脂肪酸致小鼠胰岛素抵抗的影响[J].食品科学,2015,36(1):201-204.DOI:10.7506/spkx1002-6630-201501038.
[22]RODOMAN G V,SHALAEVA T I,DOBRETSOV G E.Blood fatty acid transporters in acute pancreatitis[J].Voprosy Meditsinskoi Khimii,2001,47(6):63-66.DOI:10.1007/978-3-540-38918-7_223.
[23]JURAJ K,LIZETTE L,KAREN D,et al.Effect of liraglutide on dietary lipid induced insulin resistance in humans[J].Diabetes Obesity&Metabolism,2018,3(20):69-76.DOI:10.1111/dom.13037.
[24]韩佳彤,丁传波,顾效瑜,等.脱落酸对果糖诱导的胰岛素抵抗小鼠的影响[J].食品科学,2015,36(15):195-198.DOI:10.7506/spkx1002-6630-201515036.
[25]LI Y,HE J,LI F P,et al.The role of resistin in hepatic insulin resistance and the underlying mechanism[J].Chinese Pharmacological Bulletin,2009,25(2):190-193.DOI:10.11569/wcjd.v22.i9.1241.
[26]RALPH A,DEFRONZO D T.Skeletal muscle insulin resistance is the primay defect in type 2 diabetes[J].Diabetes Care,2009,32(2):157-160.DOI:10.2337/dc09-s302.
[27]GRUNDYS M,BREWERH B,CLEEMANJ I,et al.Definition of metabolic syndrome:report of the national heart,lung,and blood institute American heart association conference on scientific issues related to definition[J].Arteriosclerosis Thrombosis and Vascular Biology,2004,24(2):13-18.DOI:10.1161/01.atv.0000111245.75752.c6.
[28]张世卿,佟丽.胰岛素抵抗作用发生机制及实验模型的研究进展[J].中药新药与临床药理,2012,23(3):364-368.
[29]KOBAYASHI H,GAO Y H,UETAC,et al.Multilineage differentiation of cbfal deficient calvarial cells in vitro[J].Biochemical and Biophysical Research Communications,2000,273(2):630-636.DOI:10.1006/bbrc.2000.2981.
[30]姚文兵,杨红.生物化学[M].7版.北京:人民卫生出版社,2014:277-278.
[2]朱绍玲.高血脂症的社区预防与用药指导[J].中国社区医师,2013,15(3):149-151.
[3]NARA M,SUMINO H,NARA M,et al.Impaired blood rheology and elevated remnant lipoprotein particle cholesterol in hypercholesterolaemic subjects[J].Journal of International Medical Research,2009,37(2):308-310.DOI:10.1177/147323000903700204.
[4]何莲,陈邵燕,唐雯,等.慢性肾脏病与高脂血症关系的研究[J].中国实用内科杂志,2008,28(5):355-357.
[5]陈娟,邓军,张宇燕,等.丹参素对高脂血症大鼠脂代谢调节机制研究[J].中国中药杂志,2015,40(2):313-317.
[6]海静如.胰岛素抵抗与高脂血症[J].广西医学,2000,22(6):1253-1256.
[7]袁红,袁芳.单纯高脂血症与胰岛素抵抗指数的关系研究[J].中国实用内科杂志,2007,27(5):246-247.
[8]GRUNDY S M,HANSEN B,SMITH S C.Clinical management of metabolic syndrome:report of the American heart association national heart,lung,and blood institute American diabetes association conference on seientifie issues related to management[J].Circulation,2004,10(9):551-556.DOI:10.1161/01.cir.0000112379.88385.67.
[9]孔树佳,付继华.胰岛素抵抗与糖脂代谢紊乱[J].中国老年学杂志,2009,29(18):2403-2405.
[10]梁秀慈,孟文,钟英丽,等.绿原酸对高脂乳诱导小鼠胰岛素抵抗形成的影响[J].中国药理学通报,2013,29(5):654-658.
[11]冯志合,卢涛.中国柠檬酸行业概况[J].中国食品添加剂,2011,5(3):158-163.
[12]唐茂妍,陈旭东.饲用柠檬酸的应用研究进展[J].饲料与畜牧,2015,10(3):46-51.
[13]吕祥娟,刘强,宋雷,等.柠檬酸及其在猪料中的应用[J].山东畜牧兽医,2015,36(10):57-59.
[14]李珊,阿依姑丽·艾合麦提,孙鹏,等.RP-HPLC法同时测定新疆野山杏果肉中10种有机酸成分[J].食品工业科技,2017,38(20):250-255.
[15]阿依姑丽·艾合麦提,李珊,英提扎尔·艾孜木江,等.野山杏果肉总有机酸的分离纯化及其清除自由基能力的研究[J].食品科技,2017,42(5):219-224.
[16]阿依姑丽·艾合麦提,李珊,英提扎尔·艾孜木江,等.野山杏果肉总有机酸体外抑制脂质过氧化活性研究[J].西北药学杂志,2017,32(4):426-430.
[17]李珊,阿依姑丽·艾合麦提,英提扎尔·艾孜木江,等.新疆野山杏果肉总有机酸对高血脂症大鼠血清中脂质水平及抗氧化指标的影响[J].食品工业科技,2018,39(13):301-305;324.
[18]阿依姑丽·艾合麦提,英提扎尔·艾孜木江,李珊,等.野山杏果肉总有机酸对高脂血症小鼠血脂及肝脏的影响[J].华西药学杂志,2017,32(4):385-387.
[19]熊静悦,周桢昊,赵璐,等.脂肪乳剂建立高脂血症模型小鼠的比较研究[J].中医药学刊,2006,24(8):1507-1508.
[20]郑铁生,王亚娜,宗爱萍.龙虾壳聚糖干预后糖尿病小鼠血糖和糖耐量的变化[J].中国临床康复,2006,10(31):67-69.
[21]李晓月,张晶晶,张红建,等.菊粉对反式脂肪酸致小鼠胰岛素抵抗的影响[J].食品科学,2015,36(1):201-204.DOI:10.7506/spkx1002-6630-201501038.
[22]RODOMAN G V,SHALAEVA T I,DOBRETSOV G E.Blood fatty acid transporters in acute pancreatitis[J].Voprosy Meditsinskoi Khimii,2001,47(6):63-66.DOI:10.1007/978-3-540-38918-7_223.
[23]JURAJ K,LIZETTE L,KAREN D,et al.Effect of liraglutide on dietary lipid induced insulin resistance in humans[J].Diabetes Obesity&Metabolism,2018,3(20):69-76.DOI:10.1111/dom.13037.
[24]韩佳彤,丁传波,顾效瑜,等.脱落酸对果糖诱导的胰岛素抵抗小鼠的影响[J].食品科学,2015,36(15):195-198.DOI:10.7506/spkx1002-6630-201515036.
[25]LI Y,HE J,LI F P,et al.The role of resistin in hepatic insulin resistance and the underlying mechanism[J].Chinese Pharmacological Bulletin,2009,25(2):190-193.DOI:10.11569/wcjd.v22.i9.1241.
[26]RALPH A,DEFRONZO D T.Skeletal muscle insulin resistance is the primay defect in type 2 diabetes[J].Diabetes Care,2009,32(2):157-160.DOI:10.2337/dc09-s302.
[27]GRUNDYS M,BREWERH B,CLEEMANJ I,et al.Definition of metabolic syndrome:report of the national heart,lung,and blood institute American heart association conference on scientific issues related to definition[J].Arteriosclerosis Thrombosis and Vascular Biology,2004,24(2):13-18.DOI:10.1161/01.atv.0000111245.75752.c6.
[28]张世卿,佟丽.胰岛素抵抗作用发生机制及实验模型的研究进展[J].中药新药与临床药理,2012,23(3):364-368.
[29]KOBAYASHI H,GAO Y H,UETAC,et al.Multilineage differentiation of cbfal deficient calvarial cells in vitro[J].Biochemical and Biophysical Research Communications,2000,273(2):630-636.DOI:10.1006/bbrc.2000.2981.
[30]姚文兵,杨红.生物化学[M].7版.北京:人民卫生出版社,2014:277-278.