枳椇水提物对对乙酰氨基酚致小鼠急性肝损伤的相关机制研究

目的:探究枳椇水提物对对乙酰氨基酚致小鼠急性肝损伤的相关机制。方法:采用对乙酰氨基酚建立小鼠急性肝损伤模型,将大鼠分为正常组、对乙酰氨基酚(APAP)组、N-乙酰半胱氨酸(NAC)组,枳椇水提物低、中、高剂量组,计算各组小鼠的肝脏指数及肝质比指数;全自动生化仪检测血清ALT、AST、ALP水平变化;HE染色法观察小鼠肝组织病理学变化的影响;TUNEL法检测肝细胞凋亡情况;免疫组化法检测Bax、Bcl-2相关蛋白的表达;Westernblot检测Csapase-3、Caspase-9蛋白表达水平。结果:与正常组比较,APAP组小鼠肝脏重量及肝指数显著升高(P<0.01),ALT、AST、ALP水平显著升高(P<0.05),呈现明显肝组织病理损伤,TUNEL阳性细胞数升高(P<0.05),Bax蛋白表达上调(P<0.05),Bcl-2蛋白表达下调(P<0.05);与APAP组比较,枳椇水提物低、中、高剂量组小鼠肝脏重量及肝指数降低(P<0.05),ALT、AST、ALP水平显著降低(P<0.05),肝组织损伤、炎症细胞浸润明显减轻,肝组织坏死区域明显减少,TUNEL阳性细胞数减少(P<0.05),Bax蛋白表达下调(P<0.05),Bcl-2蛋白表达上调(P<0.05),Caspase-3、Caspase-9蛋白表达水平降低(P<0.05)。结论:枳椇水提物对APAP致小鼠急性肝损伤有保护作用,其作用机制可能与抑制肝细胞凋亡,下调促凋亡蛋白Bax的表达,上调抑凋亡蛋白Bcl-2的表达有关。

Study on the mechanism of water extract of Hovenia Dulcis Thunb on acetaminophen induced acute liver injury in mice

Objective:To investigate the mechanism of water extract of Hovenia Dulcis Thunb on acetaminophen induced acute liver injury in mice.Methods:The acute liver injury model of mice was established by using paracetamol.The rats were divided into normal group,acetaminophen(APAP)group,N-acetylcysteine(NAC)group and water extract of Hovenia Dulcis Thunb low,medium and high dose groups,and the liver index and liverto-mass ratio index of each group were calculated.The changes of serum ALT,AST and ALP were detected by automatic biochemical analyzer.HE staining was used to observe the effect of histopathological changes in the liver of mice.Apoptosis of hepatocytes was detected by TUNEL assay.Immunohistochemical method was used to detect the expression of Bax and Bcl-2 related proteins.Westernblot was used to detect the expression levels of Csapase-3 and Caspase-9.Results:Compared with the normal group,the liver weight and liver index of mice in the APAP group increased significantly(P<0.01),the ALT,AST and ALP levels increased significantly(P<0.05),and the pathological damage of liver tissues was obvious.The number of TUNEL positive cells increased(P<0.05),the expression of Bax protein was up-regulated(P<0.05),and the expression of Bcl-2 protein was down-regulated(P<0.05).Compared with APAP group,water extract of Hovenia Dulcis Thunb low,medium and high dose group mice liver weight and liver index decreased(P<0.05),ALT,AST,ALP level decreased significantly(P<0.05),liver tissue injury,inflammatory cells infiltration,liver tissue necrosis area decreased significantly,and the number of TUNEL positive cells decreased(P<0.05).The Bax protein expression(P<0.05),the Bcl-2 protein expression(P<0.05)and the expression levels of Caspase-3 and Caspase-9 decreased(P<0.05).Conclusion:Water extract of Hovenia Dulcis Thunb has a protective effect on acute liver injury induced by APAP in mice,and its mechanism may be related to inhibiting hepatocyte apoptosis,dow N-regulating the expression of pro-apoptotic protein Bax,and up-regulating the expression of anti-apoptotic protein Bcl-2.