基于PI3K/PKB信号通路研究吡菲尼酮缓解肝纤维化的作用机制  

Study of the mechanism of pirfenidone on relieving hepatic fibrosis based on PI3K/PKB signaling pathway

作　　者：牛向平 张素红 苏伟 吴桂红 张舒娴 NIU Xiangping;ZHANG Suhong;SU Wei;WU Guihong;ZHANG Shuxian(Electric Power Teaching Hospital,Capital Medical University,Beijing 100073,China;Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学电力教学医院,北京100073 [2]首都医科大学,北京100069

出　　处：《西北药学杂志》2020年第6期871-875,共5页Northwest Pharmaceutical Journal

基　　金：北京市教育委员会科技计划面上项目(编号:KM201510025002)。

摘　　要：目的基于磷脂酰肌醇-3-羟激酶(PI3K)/磷酸化蛋白激酶B(PKB)信号通路研究吡菲尼酮缓解小鼠肝纤维化程度的作用机制。方法选择65只小鼠,饲养7 d后将其中60只随机均分为模型组、健康组和吡菲尼酮组,每组20只,另留5只不入组。使用四氯化碳诱导模型组和吡菲尼酮组肝纤维化。成功建立肝纤维化模型后,吡菲尼酮组给予吡菲尼酮灌胃,模型组和健康组给予等量生理盐水灌胃。2周后处死小鼠,取肝脏组织进行苏木精-伊红(HE)、胶原纤维(Masson)染色,评估各组肝纤维化程度,并采用实时荧光定量聚合酶链式反应(PCR)和免疫印迹法测定PI3K和PKB的mRNA及蛋白表达量,检测小鼠血清谷草转氨酶(AST)和谷丙转氨酶(ALT)水平。结果模型组与吡菲尼酮组小鼠ALT及AST水平均明显高于健康组(P<0.05);模型组小鼠ALT及AST水平均高于吡菲尼酮组(P<0.05)。模型组小鼠PI3K mRNA和蛋白表达量明显高于吡菲尼酮组及健康组(P<0.05);吡菲尼酮组PI3K mRNA和蛋白表达量高于健康组(P<0.05);模型组小鼠PKB mRNA和蛋白表达量明显高于吡菲尼酮组及健康组(P<0.05)。模型组肝纤维化3期小鼠最多(55.56%);吡菲尼酮组2期小鼠最多(66.67%)。结论吡菲尼酮具有明显缓解小鼠肝纤维化的作用,此作用与其降低小鼠肝脏内PI3K和PKB表达量机制密切相关。Objective To investigate the mechanism of pirfenidone on relieving hepatic fibrosis in mice based on phosphatidylinositol 3-kinase/protein kinase B(PI3K/PKB)signaling pathway.Methods 60 out of 65 mice from the experimental animal center of our city were equally divided into 3 groups after feeding for 7 d,model group,healthy group and pirfenidone group,each with 20 mice,and the rest was not grouped.Carbon tetrachloride-induced hepatic fibrosis was performed in rats of model group and pirfenidone group.After the successful establishment of the hepatic fibrosis model,the pifenidone group was given pifenidone by gavage,the model group and the healthy group were given normal saline by gavage.The mice were killed 2 weeks later,liver tissues were taken for hematoxylin-eosin(HE)and Masson staining to evaluate the degree of hepatic fibrosis,the mRNA and protein expression levels of PI3K and PKB were measured by real-time fluorescence quantitative polymerase chain reaction(PCR)and Western Blot,and the serum aspartate aminotransferase(AST)and alanine aminotransferase(ALT)levels were detected.Results Serum ALT and AST levels of model group and pirafenidone group were significantly higher than those of healthy group(P<0.05).Serum ALT and AST levels of model group were significantly higher than those of pirafenidone group(P<0.05).The expression of PI3K mRNA and protein of model group was significantly higher than that of pirafenidone group and healthy group(P<0.05).The expression of PI3K mRNA and protein of pirafenidone group was significantly higher than that of healthy group(P<0.05).The expression of PKB mRNA and protein of model group was significantly higher than that of pirafenidone group and healthy group(P<0.05).The proportion of mice at stage 3 in the model group was up to 55.56%,meanwhile,the proportion of mice at stage 2 in the pirfenidone group was up to 66.67%.Conclusion Pifenidone can significantly alleviate hepatic fibrosis in mice,which is closely related to the mechanism of reducing the expression of PI3K and P

关 键 词：吡菲尼酮 磷脂酰肌醇-3-羟激酶/磷酸化蛋白激酶B 信号通路 肝纤维化 

分 类 号：R965[医药卫生—药理学]