摘要
目的:研究红花清肝十三味丸治疗急性肝损伤的作用机制。方法:48只Wistar大鼠随机分为正常对照组、模型对照组、水飞蓟宾0.1 g/kg组和红花清肝十三味丸0.25、0.5、1.0 g/kg组,1次/d,连续灌胃给药8 d。末次给药1 h后,正常对照组大鼠腹腔注射1 m1/kg花生油,其余各组注射同等体积CCl_(4-)花生油溶液建立急性肝损伤模型。生化法检测大鼠血清谷草转氨酶(AST)、谷胱甘肽(GSH)活力,Elisa法检测肝组织总抗氧化力(T-AOC)活力;TUNEL法观察大鼠肝细胞凋亡;Western blot法检测大鼠肝脏c-Jun氨基末端激酶(JNK)、磷酸化c-Jun氨基末端激酶(p-JNK)、p-c-Jun蛋白表达;Western blot法和O-RT PCR法分别检测死亡相关因子配体(FasL)、死亡相关因子(Fas)、半胱氨酸天冬氨酸蛋白酶-8(Caspase-8)、B淋巴细胞瘤-2(BCL-2)、BCL-2相关X蛋白(BAX)、细胞色素C(Cytochrome C)、半胱氨酸天冬氨酸蛋白-9(Caspase-9)的蛋白和mRNA表达。结果:与正常对照组相比,模型对照组大鼠血清AST活力显著升高(P<0.01),血清GSH及肝脏T-AOC活力显著降低(P<0.01),TUNEL染色结果显示,模型对照组小叶中心带大部肝细胞核呈阳性,可见大量凋亡小体,部分已被Kupffer细胞吞噬;模型对照组大鼠肝脏p-JNK、p-c-Jun蛋白表达及p-JNK/JNK较正常对照组升高(P<0.01),FasL、Fas、Caspase-8、Bax、Cytochrome C、Caspase-9的蛋白和mRNA表达上调,而BCL-2的蛋白和mRNA表达下调(P<0.01)。与模型对照组比较,红花清肝十三味丸各组大鼠血清AST水平,肝脏p-JNK、p-c-Jun蛋白表达,p-JNK/JNK及FasL、Fas、Bax、Cytochrome C、Caspase-8、Caspase-9的蛋白和mRNA表达均明显下调(P<0.05),而血清GSH活力明显升高及肝脏BCL-2的蛋白和mRNA表达明显上调(P<0.05);此外,红花清肝十三味丸0.5、1.0 g/kg组大鼠肝组织T-AOC活力显著升高(P<0.01);TUNEL染色显示,红花清肝十三味丸各剂量组均不同程度减少凋亡小体的形成,抑制肝细胞凋亡,且红花清肝十�
Objective:To explore the possible mechanism of Honghua Qinggan Shisanwei Pill(HQSP)in the treatment of acute liver injury.Methods:Forty-eight Wistar rats were randomly divided into the normal control group,model group,positive control(0.1 g/kg silibinin)group,and low(0.25 g/kg),medium(0.5 g/kg),and high dose(1.0 g/kg)HQSP groups,and treated with the corresponding drugs by gavage,once per day,for eight days.One hour after the last administration,rats in the normal control group were intraperitoneally injected with 1 mL/kg peanut oil,while those in the other groups were injected with the same volume of a mixture of CCl_(4) and peanut oil to induce acute liver injury.The serum glutamic oxalacetic transaminase(AST)and glutathione(GSH)activities were detected by biochemistry,and the total antioxidant capacity(T-AOC)in hepatic tissue by Elisa.The hepatocyte apoptosis was observed after TUNEL staining.The protein expression levels of c-Jun N-terminal kinase(JNK),phosphorylated JNK(p-JNK),and p-c-Jun in rat liver were measured by Western blotting.The mRNA and protein expression levels of Fas ligand(FasL),Fas,cysteine-aspartic acid protease-8(Caspase-8),B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),cytochrome C,and Caspase-9 were determined by Western blotting and O-RT PCR.Results:Compared with the normal control group,the model group showed significantly elevated serum AST activity(P<0.01)but lowered serum GSH and liver T-AOC(P<0.01).The TUNEL staining revealed that most of hepatic nuclei in the center of lobules were observed positive in the model group and a large number of apoptotic bodies were present,some of which had been phagocytized by Kupffer cells.The protein expression levels of p-JNK and p-c-Jun,p-JNK/JNK,and the mRNA and protein expression levels of FasL,Fas,Caspase-8,Bax,cytochrome C,and Caspase-9 in the model group were higher than those in the normal control group(P<0.01),while the Bcl-2 mRNA and protein expression levels were lower(P<0.01).Compared with the model group,HQSP at each dose remark
作者
卢任玲
金蓉
张子英
龚延萍
马丽杰
Lu Renling;Jin Rong;Zhang Ziying;Gong Yanping;Ma Lijie(Basic Medical College,Inner Mongolia Medical University,Hohhot 010110;Shandong Drug and Food Vocational College,Weihai 264210)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2021年第3期21-26,共6页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家自然科学基金资助项目(编号:81360676)
内蒙古自然科学基金资助项目(编号:2020MS08040)。
关键词
红花清肝十三味丸
急性肝损伤
氨基末端激酶
凋亡
Honghua Qinggan Shisanwei Pill(HQSP)
acute liver injury
c-Jun N-terminal kinase(JNK)
apoptosis
