布渣叶清热消滞退黄药效学研究
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摘要
布渣叶是广东习用地产药材,具有消食化滞、清热利湿之功效。可用于饮食积滞,感冒发热及湿热黄疸,已广泛应用于临床。关于布渣叶清热消滞退黄药效研究未见报道,本课题在中医药理论指导下,选取解热、促消化、降酶退黄、抗炎、镇痛等实验,对布渣叶清热消滞退黄药效学进行了研究,方法与结果如下：
(1)解热作用：皮下注射20%酵母混悬液8ml/kg体重的剂量7h后,大鼠体温变化升到最大值,而此时布渣叶水提物高、中剂量组与空白组比较,有显著性差异(P<0.05),且温度变化(△℃)接近正常基础体温水平；在5h段,空白组大鼠体温变化(△℃)降低到比较低的水平,而中、低剂量与空白组比较,有显著性差异(P<0.05),且体温变化(△℃)接近正常基础体温水平。实验结果提示,布渣叶水提物有比较好的解热作用,并能使干酵母致大鼠体温波段变化维持在接近正常水平。
(2)促消化作用：通过灌予营养性黑色半固体糊观察布渣叶不同提取部位及水提物、对小鼠的胃排空及小肠推进作用的影响；通过大鼠胃液分泌影响实验观察布渣叶不同提取部位对大鼠胃液量、pH值和胃蛋白酶活性的影响。①与空白组比较,香砂养胃丸组、布渣叶水提物组、乙酸乙酯部位组及剩余水层部位组均能显著减少小鼠胃内残留率,对小肠推进也有显著促进作用(P<0.05或0.01)。②与空白组比较,香砂养胃丸组、正丁醇部位组及剩余水层部位组均能显著增加大鼠胃液量(P<0.05)；香砂养胃丸组、乙酸乙酯组、正丁醇组及剩余水层组均能显著降低胃液pH值(P<0.05或0.01)；各组中仅正丁醇组能显著提高胃蛋白酶活性(P<0.01)。实验结果提示,布渣叶可通过降低胃内残留率、促进小肠推进、增加胃液分泌量和胃液酸度及提高胃蛋白酶活性达到促消化作用。布渣叶水提物、乙酸乙酯部位、正丁醇部位和剩余水层部位均含有药效成分。
(3)降酶退黄作用：采用α-萘异硫氰酸酯(ANIT)致小鼠胆汁瘀积肝损伤模型,观察布渣叶对模型小鼠血清总胆红素(T-BIL)含量、碱性磷酸酶(ALP)活性、门冬氨酸转氨酶(AST)活性、丙氨酸转氨酶(ALT)活性的影响。①与模型对照组比较,布渣叶各剂量组均能显著降低ANIT所致黄疸模型小鼠血清中T-BIL与D-BIL的含量,并且降低程度基本接近正常对照组,并能显著抑制ALP、AST和ALT的酶活性(P<0.01),说明布渣叶水提物有明显的退黄和改善肝功能的作用。②各给药组与模型对照组比较,茵栀黄组和剩余水层部位组能显著降低ANIT致胆汁瘀积模型小鼠血清中T-BIL的含量、ALP活性、AST活性、ALT活性及肝脏指数(P<0.05或0.01)；布渣叶水提物组和正丁醇部位组能显著降低血清中T-BIL的含量、AST活性、ALT活性及肝脏脏器指数(P<0.05或0.01)；石油醚部位组和乙酸乙酯部位组对血清T-BIL的含量、ALP活性、AST活性、ALT活性及肝脏指数在统计学上均无明显差异(P>0.05)。表明布渣叶水提物、正丁醇部位及各有机溶剂萃取后的剩余水层部位具有明显的降酶退黄作用,而其他部位如石油醚部位、乙酸乙酯部位则无降酶退黄作用。实验结果提示,布渣叶可通过降低胆汁瘀积模型小鼠T-BIL含量、抑制ALP、AST、ALT酶活性发挥降酶退黄作用,其药效部位主要存在于正丁醇部位和剩余水层部位。
(4)抗炎作用：通过二甲苯致小鼠耳廓肿胀实验及腹腔注射醋酸致小鼠腹腔毛细血管通透性增高实验,观察布渣叶水提物对炎症反应的影响。实验结果表明,布渣叶水提物高、中剂量组对二甲苯引起的小鼠耳廓肿胀有明显抑制作用(P<0.05或0.01)；布渣叶水提物高、中、低剂量组均能显著抑制由醋酸引起的组织毛细血管的通透性增加(P<0.05)。实验结果提示,布渣叶水提物具有显著的抗急性炎症作用。
(5)镇痛作用：采用小鼠热板法和冰醋酸致小鼠扭体反应,观察布渣叶水提物对疼痛的抑制作用。结果表明,布渣叶水提物各剂量组均能显著抑制小鼠因热刺激所引起的疼痛反应(P<0.05或0.01)；而高、低剂量组能显著抑制小鼠因化学刺激所引起的疼痛反应(P<0.05或0.01)。实验结果提示,布渣叶水提物具有较好的镇痛作用。
Folium Microcotis in Guangdong is a conventional native drug, with the effect of relieving dyspepsia and clearing away heat evil and promoting diuresis, can be used for dyspeptic retention, fever caused by exogenous pathogens and jaundice with damp-heat pathogen, has been widely used in clinical. About the study on Pharmacodynamics of Relieve Fever, Promoting Digestion and Reducing Jaundice of Folium Microcotis were no reports on efficacy, the subject selected antipyretic, promoting digestion, reducing enzyme and reducing jaundice, anti-inflammatory, analgesic and other experiments, under the guidance of the theory of Traditional Chinese Medicine, methods and results are as follows:
(1) Antipyretic:Subcutaneous injection of 20% of body weight dose of yeast suspension 8ml/kg for 7h, the rat body temperature had rise to maximum, at this time the high dose group and the middle dose group of aqueous extract from Folium Microcotis compared with the control group, there was significant difference(P＜0.05), and the temperature change (△℃) to near normal levels of basal body temperature; In paragraph 5h, blank rat body temperature change (△℃) reduce to a relatively low level, while the low dose and blank group, there were significant differences (P<0.05), and the temperature change (△℃) basal body temperature close to normal levels. The results suggest that aqueous extract Folium Microcotis have better antipyretic effect and can dry yeast-induced changes in body temperature band remained at near normal levels.
(2) Promote digestion:With irrigating the black nutritional semi-solid paste, observing the effects of the different extracts from Folium Microcotis on gastric emptying and the propulsive ratio of mice's small intestine; Through the experiment affected by gastric juice secretion in rats, observing the different Folium Microcotis extracts on gastric juice volume, pH, and pepsin activity.①Compared with the control group, Xiangshayangwei pill group, aqueous extract group of Folium Microcotis, ethyl acetate group and surplus water group can significantly reduce the area group residual rate of the mice stomach, small intestine also on a significant role in promoting (P < 0.05 or 0.01).②Compared with the control group, Xiangshayangwei pill group, n-butanol and surplus water group can increase gastric juice volume was significantly (P< 0.05); Xiangshayangwei pill group, ethyl group, n-butyl alcohol group and surplus water group can significantly reduce the gastric juice pH values (P<0.05 or 0.01); Each group, only n-butanol could significantly increase pepsin activity (P<0.01). These results suggest Folium Microcotis can reduce the gastric emptying rate, the promotion of small intestine, increased gastric secretion, decreased gastric acidity and pepsin activity to enhance and promote digestion. The aqueous extract, ethyl acetate extract, n-butanol extract and the remaining parts of the water layer of Folium Microcotis contained the efficacy component parts.
(3) Reducing enzyme and reducing jaundice:Usingα-naphthyl isothiocyan-ate (ANIT) induced the mice model of cholestasis liver injury, observing the extracts of Folium Microcotis on total bilirubin(T-BIL) levels, alkaline phosphatase (ALP) activity, aspartate aminotransferase (AST) activity and alanine aminotransferase (ALT) activity in the serum of the mice model.①With the model control group, all groups of Folium Microcotis can significantly reduce the serum levels of T-BIL and D-BIL on model animal, and lower level close to the normal control group, and significantly inhibited ALP, AST and ALT enzyme activity (P＜0.01), that aqueous extract from Folium Microcotis had obvious effect on reducing enzyme and receding jaundice.②The treatment group compare with model group, Yinzhihuang group and surplus water group could significantly reduce the ANIT induced cholestasis model of serum levels of T-BIL, ALP activity, AST activity, ALT activity and liver index (P＜0.05 or 0.01); petroleum group and ethyl acetate group were no statistically significant difference (P＞0.05). Showing that aqueous extract, n-butanol and the part of surplus water layer with significant role on reducing enzyme and reducing jaundice, while other parts, such as petroleum ether, ethyl acetate extract had no reducing the enzyme jaundice effect. These results suggest the active constituents of Folium Microcotis exist in surplus water extract and surplus water extract.
(4) Anti-inflammatory effect:Various inflammatory models, including the swelling of ear induced by xylene in mice, the peimeability increase of Capillary permeability by acetic acid in mice were used to explore the anti-inflammatory of Water-extract of Folium Microcotis. The results showed that high and middle doses of Folium Microcotis could significantly inhibit ear edema(P＜0.05 or 0.01), and all the three doses of Folium Microcotis could significantly inhibit the peimeability increase of Capillary permeability (P＜0.05). The results suggest that aqueous extract of Folium Microcotis has significant effect against acute inflammation.
(5) The analgesic effect:The mice hot plate and acetic acid induced writhing response observed Folium Microcotis aqueous extract of pain inhibition. The results showed that water extract of Folium Microcotis can each dose group was significantly inhibited in mice due to thermal stimulation caused by pain response (P<0.05 or 0.01); the high and low dose group was significantly inhibited in mice caused by chemical stimulation pain response (P＜0.05 or 0.01). The results suggest that aqueous extract of Folium Microcotis has good analgesic effect.
(1)解热作用：皮下注射20%酵母混悬液8ml/kg体重的剂量7h后,大鼠体温变化升到最大值,而此时布渣叶水提物高、中剂量组与空白组比较,有显著性差异(P<0.05),且温度变化(△℃)接近正常基础体温水平；在5h段,空白组大鼠体温变化(△℃)降低到比较低的水平,而中、低剂量与空白组比较,有显著性差异(P<0.05),且体温变化(△℃)接近正常基础体温水平。实验结果提示,布渣叶水提物有比较好的解热作用,并能使干酵母致大鼠体温波段变化维持在接近正常水平。
(2)促消化作用：通过灌予营养性黑色半固体糊观察布渣叶不同提取部位及水提物、对小鼠的胃排空及小肠推进作用的影响；通过大鼠胃液分泌影响实验观察布渣叶不同提取部位对大鼠胃液量、pH值和胃蛋白酶活性的影响。①与空白组比较,香砂养胃丸组、布渣叶水提物组、乙酸乙酯部位组及剩余水层部位组均能显著减少小鼠胃内残留率,对小肠推进也有显著促进作用(P<0.05或0.01)。②与空白组比较,香砂养胃丸组、正丁醇部位组及剩余水层部位组均能显著增加大鼠胃液量(P<0.05)；香砂养胃丸组、乙酸乙酯组、正丁醇组及剩余水层组均能显著降低胃液pH值(P<0.05或0.01)；各组中仅正丁醇组能显著提高胃蛋白酶活性(P<0.01)。实验结果提示,布渣叶可通过降低胃内残留率、促进小肠推进、增加胃液分泌量和胃液酸度及提高胃蛋白酶活性达到促消化作用。布渣叶水提物、乙酸乙酯部位、正丁醇部位和剩余水层部位均含有药效成分。
(3)降酶退黄作用：采用α-萘异硫氰酸酯(ANIT)致小鼠胆汁瘀积肝损伤模型,观察布渣叶对模型小鼠血清总胆红素(T-BIL)含量、碱性磷酸酶(ALP)活性、门冬氨酸转氨酶(AST)活性、丙氨酸转氨酶(ALT)活性的影响。①与模型对照组比较,布渣叶各剂量组均能显著降低ANIT所致黄疸模型小鼠血清中T-BIL与D-BIL的含量,并且降低程度基本接近正常对照组,并能显著抑制ALP、AST和ALT的酶活性(P<0.01),说明布渣叶水提物有明显的退黄和改善肝功能的作用。②各给药组与模型对照组比较,茵栀黄组和剩余水层部位组能显著降低ANIT致胆汁瘀积模型小鼠血清中T-BIL的含量、ALP活性、AST活性、ALT活性及肝脏指数(P<0.05或0.01)；布渣叶水提物组和正丁醇部位组能显著降低血清中T-BIL的含量、AST活性、ALT活性及肝脏脏器指数(P<0.05或0.01)；石油醚部位组和乙酸乙酯部位组对血清T-BIL的含量、ALP活性、AST活性、ALT活性及肝脏指数在统计学上均无明显差异(P>0.05)。表明布渣叶水提物、正丁醇部位及各有机溶剂萃取后的剩余水层部位具有明显的降酶退黄作用,而其他部位如石油醚部位、乙酸乙酯部位则无降酶退黄作用。实验结果提示,布渣叶可通过降低胆汁瘀积模型小鼠T-BIL含量、抑制ALP、AST、ALT酶活性发挥降酶退黄作用,其药效部位主要存在于正丁醇部位和剩余水层部位。
(4)抗炎作用：通过二甲苯致小鼠耳廓肿胀实验及腹腔注射醋酸致小鼠腹腔毛细血管通透性增高实验,观察布渣叶水提物对炎症反应的影响。实验结果表明,布渣叶水提物高、中剂量组对二甲苯引起的小鼠耳廓肿胀有明显抑制作用(P<0.05或0.01)；布渣叶水提物高、中、低剂量组均能显著抑制由醋酸引起的组织毛细血管的通透性增加(P<0.05)。实验结果提示,布渣叶水提物具有显著的抗急性炎症作用。
(5)镇痛作用：采用小鼠热板法和冰醋酸致小鼠扭体反应,观察布渣叶水提物对疼痛的抑制作用。结果表明,布渣叶水提物各剂量组均能显著抑制小鼠因热刺激所引起的疼痛反应(P<0.05或0.01)；而高、低剂量组能显著抑制小鼠因化学刺激所引起的疼痛反应(P<0.05或0.01)。实验结果提示,布渣叶水提物具有较好的镇痛作用。
Folium Microcotis in Guangdong is a conventional native drug, with the effect of relieving dyspepsia and clearing away heat evil and promoting diuresis, can be used for dyspeptic retention, fever caused by exogenous pathogens and jaundice with damp-heat pathogen, has been widely used in clinical. About the study on Pharmacodynamics of Relieve Fever, Promoting Digestion and Reducing Jaundice of Folium Microcotis were no reports on efficacy, the subject selected antipyretic, promoting digestion, reducing enzyme and reducing jaundice, anti-inflammatory, analgesic and other experiments, under the guidance of the theory of Traditional Chinese Medicine, methods and results are as follows:
(1) Antipyretic:Subcutaneous injection of 20% of body weight dose of yeast suspension 8ml/kg for 7h, the rat body temperature had rise to maximum, at this time the high dose group and the middle dose group of aqueous extract from Folium Microcotis compared with the control group, there was significant difference(P＜0.05), and the temperature change (△℃) to near normal levels of basal body temperature; In paragraph 5h, blank rat body temperature change (△℃) reduce to a relatively low level, while the low dose and blank group, there were significant differences (P<0.05), and the temperature change (△℃) basal body temperature close to normal levels. The results suggest that aqueous extract Folium Microcotis have better antipyretic effect and can dry yeast-induced changes in body temperature band remained at near normal levels.
(2) Promote digestion:With irrigating the black nutritional semi-solid paste, observing the effects of the different extracts from Folium Microcotis on gastric emptying and the propulsive ratio of mice's small intestine; Through the experiment affected by gastric juice secretion in rats, observing the different Folium Microcotis extracts on gastric juice volume, pH, and pepsin activity.①Compared with the control group, Xiangshayangwei pill group, aqueous extract group of Folium Microcotis, ethyl acetate group and surplus water group can significantly reduce the area group residual rate of the mice stomach, small intestine also on a significant role in promoting (P < 0.05 or 0.01).②Compared with the control group, Xiangshayangwei pill group, n-butanol and surplus water group can increase gastric juice volume was significantly (P< 0.05); Xiangshayangwei pill group, ethyl group, n-butyl alcohol group and surplus water group can significantly reduce the gastric juice pH values (P<0.05 or 0.01); Each group, only n-butanol could significantly increase pepsin activity (P<0.01). These results suggest Folium Microcotis can reduce the gastric emptying rate, the promotion of small intestine, increased gastric secretion, decreased gastric acidity and pepsin activity to enhance and promote digestion. The aqueous extract, ethyl acetate extract, n-butanol extract and the remaining parts of the water layer of Folium Microcotis contained the efficacy component parts.
(3) Reducing enzyme and reducing jaundice:Usingα-naphthyl isothiocyan-ate (ANIT) induced the mice model of cholestasis liver injury, observing the extracts of Folium Microcotis on total bilirubin(T-BIL) levels, alkaline phosphatase (ALP) activity, aspartate aminotransferase (AST) activity and alanine aminotransferase (ALT) activity in the serum of the mice model.①With the model control group, all groups of Folium Microcotis can significantly reduce the serum levels of T-BIL and D-BIL on model animal, and lower level close to the normal control group, and significantly inhibited ALP, AST and ALT enzyme activity (P＜0.01), that aqueous extract from Folium Microcotis had obvious effect on reducing enzyme and receding jaundice.②The treatment group compare with model group, Yinzhihuang group and surplus water group could significantly reduce the ANIT induced cholestasis model of serum levels of T-BIL, ALP activity, AST activity, ALT activity and liver index (P＜0.05 or 0.01); petroleum group and ethyl acetate group were no statistically significant difference (P＞0.05). Showing that aqueous extract, n-butanol and the part of surplus water layer with significant role on reducing enzyme and reducing jaundice, while other parts, such as petroleum ether, ethyl acetate extract had no reducing the enzyme jaundice effect. These results suggest the active constituents of Folium Microcotis exist in surplus water extract and surplus water extract.
(4) Anti-inflammatory effect:Various inflammatory models, including the swelling of ear induced by xylene in mice, the peimeability increase of Capillary permeability by acetic acid in mice were used to explore the anti-inflammatory of Water-extract of Folium Microcotis. The results showed that high and middle doses of Folium Microcotis could significantly inhibit ear edema(P＜0.05 or 0.01), and all the three doses of Folium Microcotis could significantly inhibit the peimeability increase of Capillary permeability (P＜0.05). The results suggest that aqueous extract of Folium Microcotis has significant effect against acute inflammation.
(5) The analgesic effect:The mice hot plate and acetic acid induced writhing response observed Folium Microcotis aqueous extract of pain inhibition. The results showed that water extract of Folium Microcotis can each dose group was significantly inhibited in mice due to thermal stimulation caused by pain response (P<0.05 or 0.01); the high and low dose group was significantly inhibited in mice caused by chemical stimulation pain response (P＜0.05 or 0.01). The results suggest that aqueous extract of Folium Microcotis has good analgesic effect.
引文
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