抗纤软肝颗粒对肝纤维化小鼠PI3K/Akt/mTOR信号通路的作用  

Effect of Kangxian Ruangan Granules(抗纤软肝颗粒)on the Expression of PI3K/Akt/mTOR Signaling Pathway in Mice with Liver Fibrosis

作　　者：徐俊 戚璐[1] 郑新平 周涛[1] 李硕 胡聪 程良斌[1] XU Jun;QI Lu;ZHENG Xin-ping;ZHOU Tao;LI Shuo;HU Cong;CHENG Liang-bin(Hubei University of Traditional Chinese Medicine,Wuhan Hubei 430065,China;Jingzhou Hospital of Traditional Chinese Medicine,Jingzhou Hubei 434000,China)

机构地区：[1]湖北中医药大学,湖北武汉430065 [2]荆州市中医医院,湖北荆州434000

出　　处：《中医药导报》2020年第14期4-8,共5页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家中医药管理局国家中医临床研究基地业务建设科研专项(JDZX2015179)。

摘　　要：目的:观察抗纤软肝颗粒对CCl4诱导肝纤维化小鼠磷酸肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路的影响,探讨抗纤软肝颗粒防治肝纤维化的可能机制。方法:从45只小鼠中随机选取10只作为对照组,其余小鼠皮下注射40%CCl4橄榄油溶液,每3 d一次,持续8周,诱导肝纤维化小鼠模型。将造模成功小鼠随机分为模型组、抗纤软肝颗粒组和秋水仙碱组。于第9周,各给药组分别灌胃相应的受试药物,1次/d,给药8周。实验结束后,计算各组小鼠体质量变化、肝脏指数;对肝组织进行HE和Masson染色,免疫组化法检测肝组织PI3K、Akt表达,Western blotting检测mTOR表达;RT-qPCR检测肝组织PI3K mRNA、Akt mRNA、mTOR mRNA表达。结果:HE和Masson染色显示,模型组小鼠的肝组织中胶原纤维广泛增生,纤维间隔相互连接,假小叶形成;抗纤软肝颗粒组与秋水仙碱组小鼠的肝脏组织纤维化程度较模型组减轻。免疫组化结果显示,模型组小鼠肝组织PI3K、Akt表达明显高于对照组(P<0.05),抗纤软肝颗粒组与秋水仙碱组的肝组织PI3K、Akt的表达相较于模型组明显下降(P<0.05)。Western blotting结果显示,模型组小鼠肝组织mTOR高于对照组(P<0.05),抗纤软肝颗粒组与秋水仙碱组小鼠肝组织mTOR的表达相较于模型组明显下降(P<0.05)。RT-qPCR显示,模型组小鼠肝脏PI3K mRNA、Akt mRNA、mTOR mRNA表达明显高于对照组(P<0.05),而抗纤软肝颗粒组与秋水仙碱组小鼠肝脏PI3K mRNA、Akt mRNA、mTOR mRNA表达较模型组下降(P<0.05)。结论:抗纤软肝颗粒对肝纤维化小鼠具有一定的治疗作用,其机制可能与抑制PI3K/Akt/mTOR信号通路有关。Objective:To observe the effect of Kangxian Ruangan granules on phosphoinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian rapamycin target protein(mTOR)signaling pathway in CCl4-induced liver fibrosis mice,and to explore the possible mechanism of Kangxian Ruangan granules in preventing and treating liver fibrosis.Methods:10 mice were randomly selected as the control group,and the remaining mice were subcutaneously injected with 40%CCl4 olive oil solution every 3 days for 8 weeks to induce liver fibrosis in mice.The successfully modeled mice were randomly divided into model group,Kangxian Ruangan granules group and colchicine group.In the 9th week,each administration group was given the corresponding test drug by gavage,once daily,for 8 weeks.After the experiment,the weight changes and liver index of each group of mice were calculated,the liver tissues were stained with HE and Masson,the expression of PI3K and Akt in the liver tissues were detected by immunohistochemistry,and the expression of mTOR was detected by Western blotting.The expression of PI3K mRNA,akt mRNA and mTOR mRNA in liver tissue was detected by RT-qPCR.Results:HE and Masson staining showed that the collagen fibers in the liver tissues of the model group were extensively proliferated,the fibrous septum were interconnected,and pseudolobules were formed;the degree of liver fibrosis in Kangxian Ruangan granules group and colchicine group was less than that in the model group.The results of immunohistochemistry showed that the expression of PI3K and Akt in the liver tissue of the model group was significantly higher than that of the control group(P<0.05),and the expression of PI3K and Akt in the liver tissues of the Kangxian Ruangan granules group and the colchicine group were significantly lower than that of the model group(P<0.05).Western blotting results showed that mTOR in liver tissue of mice in the model group was higher than that in the control group(P<0.05),and the expression of mTOR in liver tissue of mice in the Kangxian Ruangan granule

关 键 词：抗纤软肝颗粒 肝纤维化 磷酸肌醇3-激酶 蛋白激酶B 小鼠 

分 类 号：R285.5[医药卫生—中药学]