藏药苦空确屯卡察尔保肝作用研究及机制探讨
摘要
随着社会经济发展,人民生活水平的不断提高,各种类型肝病,包括病毒性肝炎、脂肪肝、酒精性肝病等急慢性肝病的发病率在全球范围内呈上升趋势,已经严重威胁到全球人类的健康。尤其是病毒性肝炎,由于至今世界尚无一种药物能真正起到根治作用,开发和筛选对肝脏有确切保护作用的新药,仍是一项艰巨的工作。
目的：苦空确屯卡察尔(ZYJN)是在藏医经验方“七味红花殊胜丸”基础上加减而成,临床上拟用于各种急、慢性肝炎,新旧肝病的治疗,本课题着重探讨其保肝降酶、利胆退黄作用及作用机制。
方法：①腹腔注射D-Galn造成小鼠急性肝损伤模型,测定小鼠肝脏指数及血清ALT、AST水平,并对肝组织做HE染色,观察ZYJN对急性肝损伤的影响；皮下注射CCL_4造成大鼠慢性肝损伤模型,测定大鼠肝脏系数及血清ALT、AST、TP、ALB、T-BIL、LN、HA水平以及肝组织匀浆MDA和Hyp含量,做肝组织HE染色和免疫组化染色,观察ZYJN对慢性肝损伤的影响；②大鼠胆总管插管,观察ZYJN对正常大鼠胆汁流量的影响；腹腔注射ANIT造成小鼠胆汁淤积性黄疸,观察ZYJN的退黄功效；③腹腔注射环磷酰胺造成小鼠免疫低下模型,观察ZYJN对免疫功能的影响。
结果：①ZYJN能明显降低D-Galn致小鼠血清ALT、AST水平及肝脏指数,明显改善急性肝损伤小鼠肝脏病理变化；明显降低CCL4致慢性肝损伤大鼠肝脏指数以及血清ALT、AST、LN、HA水平和肝组织MDA、Hyp的含量,升高血清TP水平,并能明显改善慢性肝损伤大鼠肝脏病理变化,抑制肝组织对a-SMA、TGF-β1的表达,但是对MMP-1的表达无明显影响：②ZYJN单次给药能够明显增加大鼠胆汁流量,在给药0.5h后作用明显,维持6h；能明显降低ANIT所致胆汁淤积性黄疸小鼠血清T-BIL、ALP、ALT、AST水平；③ZYJN能明显增强环磷酰胺致免疫力低下小鼠巨噬细胞对刚果红的吞噬能力,增强小鼠免疫力：
结论：苦空确屯卡察尔具有较好的保肝降酶、利胆退黄作用,其作用机制可能与以下几方面相关：①通过降低细胞膜通透性、抑制细胞膜脂质过氧化、减轻肝纤维化程度、改善肝组织病理变化从而对急、慢性肝损伤起到保护作用；②通过促进胆汁分泌、保护胆管上皮细胞、减少炎症发生起到退黄利胆的作用；③通过增强单核巨噬细胞的吞噬作用起到增强非特异性免疫功能的作用。
Object:To study the hepatic-protective effects and mechanism of "kukongquet-unkachaer"(ZYJN) which was the empirical formula of tibetan medicine and used for various kinds of acute hepatitis, chronic hepatitis clinically.
Metherd:①D-Galn was given to the mouse by intraperitoneal injection to establish acute liver injury model. The hepatosomatic index of the mouse、the alanine aminotransferase(ALT), aspartate aminotransferase (AST) levels in serum were all detected, and the liver pathological injury were observed after HE staining. CCL4 was given to the rat by subcutaneous injection to establish chronic liver injury models. The ALT,AST,total protein(TP),albumin(ALB),total bilirubin(T-BIL), laminin(LN), Hyaluronic(HA), levels in serum, hepatosomatic index and the malonaldehyde(MDA), hydroxyproline(Hyp) contents in liver homogenate were determined, the liver pathological injury was observed after HE staining and immunohistochemistry staining. All above were to research the protective effect of ZYJN on the liver. A pipe was intubated into the common bile duct of the rat, and bile flow of the rat was observed to research the affection of ZYJN on the bile flow; a-isothiocyanate(ANIT) was given to the mouse by intraperitoneal injection to made cholestatic jaundice, and the effects of ZYJN on jaundice-relieving was observed. Cyclophosphamide was given to the mouse by intraperitoneal injection to establish hyp-immune models, the effects of ZYJN on immune function was observed.
Results:①In mouse acute liver injury models induced by D-Galn,ZYJN significantly decreased ALT,AST levels in serum and improved the morphological changes; In rat chronic liver injury models induced by CCL4, ZYJN significantly decreased ALT,AST,LN,HA levels in serum,MDA,Hyp contents in liver homogenate, increased TP levels in serum. Besides, ZYJN significantly improved morphological changes and decrease the expression of a-SMA and TGF-β1 in liver tissue.②ZYJN can improve the excretion of rat's bile after single dosage especially in 0.5h, decrease significantly the T-BIL,ALP, ALT,AST levels in serum;③ZYJN enhanced the function of monocytes in hypoimmunologic model mouse.
Conclusion:ZYJN has the function on protecting acute and chronic liver injury, the mechanism may be as follows:①By diminutus membrane permeability suppress-ing lipid peroxidation of cell membrane, lightening the level of hepatic fibrosis, improving the patho-state of hepatic tissue, ZYJN protected acute and chronic liver injury.②YJN has the good pharmacological effects on promoting bile secretion and reducing jaundice;③ZYJN enhanced immunity by strengthen phagocytosis of mononuclear macrophage.
目的：苦空确屯卡察尔(ZYJN)是在藏医经验方“七味红花殊胜丸”基础上加减而成,临床上拟用于各种急、慢性肝炎,新旧肝病的治疗,本课题着重探讨其保肝降酶、利胆退黄作用及作用机制。
方法：①腹腔注射D-Galn造成小鼠急性肝损伤模型,测定小鼠肝脏指数及血清ALT、AST水平,并对肝组织做HE染色,观察ZYJN对急性肝损伤的影响；皮下注射CCL_4造成大鼠慢性肝损伤模型,测定大鼠肝脏系数及血清ALT、AST、TP、ALB、T-BIL、LN、HA水平以及肝组织匀浆MDA和Hyp含量,做肝组织HE染色和免疫组化染色,观察ZYJN对慢性肝损伤的影响；②大鼠胆总管插管,观察ZYJN对正常大鼠胆汁流量的影响；腹腔注射ANIT造成小鼠胆汁淤积性黄疸,观察ZYJN的退黄功效；③腹腔注射环磷酰胺造成小鼠免疫低下模型,观察ZYJN对免疫功能的影响。
结果：①ZYJN能明显降低D-Galn致小鼠血清ALT、AST水平及肝脏指数,明显改善急性肝损伤小鼠肝脏病理变化；明显降低CCL4致慢性肝损伤大鼠肝脏指数以及血清ALT、AST、LN、HA水平和肝组织MDA、Hyp的含量,升高血清TP水平,并能明显改善慢性肝损伤大鼠肝脏病理变化,抑制肝组织对a-SMA、TGF-β1的表达,但是对MMP-1的表达无明显影响：②ZYJN单次给药能够明显增加大鼠胆汁流量,在给药0.5h后作用明显,维持6h；能明显降低ANIT所致胆汁淤积性黄疸小鼠血清T-BIL、ALP、ALT、AST水平；③ZYJN能明显增强环磷酰胺致免疫力低下小鼠巨噬细胞对刚果红的吞噬能力,增强小鼠免疫力：
结论：苦空确屯卡察尔具有较好的保肝降酶、利胆退黄作用,其作用机制可能与以下几方面相关：①通过降低细胞膜通透性、抑制细胞膜脂质过氧化、减轻肝纤维化程度、改善肝组织病理变化从而对急、慢性肝损伤起到保护作用；②通过促进胆汁分泌、保护胆管上皮细胞、减少炎症发生起到退黄利胆的作用；③通过增强单核巨噬细胞的吞噬作用起到增强非特异性免疫功能的作用。
Object:To study the hepatic-protective effects and mechanism of "kukongquet-unkachaer"(ZYJN) which was the empirical formula of tibetan medicine and used for various kinds of acute hepatitis, chronic hepatitis clinically.
Metherd:①D-Galn was given to the mouse by intraperitoneal injection to establish acute liver injury model. The hepatosomatic index of the mouse、the alanine aminotransferase(ALT), aspartate aminotransferase (AST) levels in serum were all detected, and the liver pathological injury were observed after HE staining. CCL4 was given to the rat by subcutaneous injection to establish chronic liver injury models. The ALT,AST,total protein(TP),albumin(ALB),total bilirubin(T-BIL), laminin(LN), Hyaluronic(HA), levels in serum, hepatosomatic index and the malonaldehyde(MDA), hydroxyproline(Hyp) contents in liver homogenate were determined, the liver pathological injury was observed after HE staining and immunohistochemistry staining. All above were to research the protective effect of ZYJN on the liver. A pipe was intubated into the common bile duct of the rat, and bile flow of the rat was observed to research the affection of ZYJN on the bile flow; a-isothiocyanate(ANIT) was given to the mouse by intraperitoneal injection to made cholestatic jaundice, and the effects of ZYJN on jaundice-relieving was observed. Cyclophosphamide was given to the mouse by intraperitoneal injection to establish hyp-immune models, the effects of ZYJN on immune function was observed.
Results:①In mouse acute liver injury models induced by D-Galn,ZYJN significantly decreased ALT,AST levels in serum and improved the morphological changes; In rat chronic liver injury models induced by CCL4, ZYJN significantly decreased ALT,AST,LN,HA levels in serum,MDA,Hyp contents in liver homogenate, increased TP levels in serum. Besides, ZYJN significantly improved morphological changes and decrease the expression of a-SMA and TGF-β1 in liver tissue.②ZYJN can improve the excretion of rat's bile after single dosage especially in 0.5h, decrease significantly the T-BIL,ALP, ALT,AST levels in serum;③ZYJN enhanced the function of monocytes in hypoimmunologic model mouse.
Conclusion:ZYJN has the function on protecting acute and chronic liver injury, the mechanism may be as follows:①By diminutus membrane permeability suppress-ing lipid peroxidation of cell membrane, lightening the level of hepatic fibrosis, improving the patho-state of hepatic tissue, ZYJN protected acute and chronic liver injury.②YJN has the good pharmacological effects on promoting bile secretion and reducing jaundice;③ZYJN enhanced immunity by strengthen phagocytosis of mononuclear macrophage.
引文
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