基于IRS-PI3K-AKT-mTOR 信号通路探讨祛痰活血方治疗非酒精性脂肪性肝病大鼠的机制  

Investigation of the mechanism of action of Qutan Huoxue decoction in treating non-alcoholic fatty liver disease in rats based on the IRS-PI3K-AKT-mTOR signaling pathway

作　　者：曾勇 彭孟云 张玉蓉 汪静 ZENG Yong;PENG Mengyun;ZHANG Yurong;WANG Jing(Department of Hepatobiliary Diseases,the Affiliated T.C.M Hospital of Southwest Medical University,Luzhou 646000,Sichuan Province,China)

机构地区：[1]西南医科大学附属中医医院肝胆病科,四川泸州646000

出　　处：《西南医科大学学报》2020年第4期323-328,共6页Journal of Southwest Medical University

基　　金：泸州市人民政府-西南医科大学高层次人才(陈晨团队)引进专项资助;四川省教育厅项目(18ZB0652);泸州市科技计划项目[2017-S-41(5/7)]。

摘　　要：目的:利用非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)大鼠模型探讨祛痰活血方通过调控IRS(胰岛素受体底物)/PI3K(磷脂酰肌醇3激酶)/AKT(蛋白激酶B)/mTOR(雷帕霉素靶蛋白)信号通路治疗NAFLD的机制。方法:选择健康雄性SD大鼠30只适应性喂养1周后,随机分为正常组(10只)、造模组(20只),正常组予普通饲料,造模组予高脂饲料建模,时间共计8周,验证造模成功后,将剩余造模组(18只)随机分为模型组(9只)、祛痰活血方组(9只)。正常组、模型组予以生理盐水10 mL/(kg·d)灌胃,祛痰活血方组予以祛痰活血方生药11.7 g/(kg·d)灌胃。干预4周后处死大鼠,试剂盒检测血清总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、空腹血糖(FBG),ELISA法检测大鼠空腹血清胰岛素(FINS)、极低密度脂蛋白(VLDL),计算胰岛素抵抗指数(HOMA-IR)。Western-Blot法检测肝脏组织IRS-1、PI3K、p-PI3K、AKT、p-AKT、mTORC1蛋白表达水平。采用苏木精-伊红(HE)及油红O染色观察各组大鼠肝脏组织形态学改变及脂肪变程度。结果:与正常组相比,模型组TC、TG、LDL-C、VLDL明显升高,HDL-C降低,FBG、FINS、HOMA-IR明显升高,肝脏组织中IRS-1、p-PI3K、p-AKT、mTORC1蛋白表达升高,差异均有统计学意义(P<0.05);与模型组相比,祛痰活血方能显著降低TC、TG、LDL-C、VLDL,升高HDL-C,降低FBG、FINS、HOMA-IR,降低肝脏组织中IRS-1、p-PI3K、p-AKT、mTORC1蛋白表达,差异均有统计学意义(P<0.05);与模型组比较,祛痰活血方能显著改善NAFLD大鼠肝脏脂质沉积和脂肪变程度。结论:祛痰活血方可通过调节IRS/PI3K/AKT/mTOR信号通路改善胰岛素抵抗,调节糖脂代谢,改善NAFLD大鼠肝脏脂肪变性程度。Objective:To explore the therapeutic mechanism of Qutan Huoxue decoction in a rat model of non-alcoholic fatty liver disease(NAFLD)based on the insulin receptor substrate(IRS)/phosphatidylinositol 3-ki⁃nase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)signaling pathway.Methods:Thirty healthy male Sprague-Dawley rats were selected for adaptive feeding for 1 week and randomly divided into normal group(n=10)and modeling group(n=20).The normal group was given normal diet and the modeling group was giv⁃en high-fat diet for 8 weeks.After the modeling was verified to be successful,the remaining rats in the modeling group(n=18)were randomly divided into model group(n=9)and Qutan Huoxue decoction group(n=9).The nor⁃mal group and the model group were given normal saline by gavage at a dose of 10 mL/(kg·d),and the Qutan Huoxue decoction group was given intragastric administration of the Qutan Huoxue crude drug at a dose of 11.7 g/(kg·d).The rats were sacrificed after 4 weeks of intervention.Kits were used to measure serum total cholesterol(TC),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),and fasting blood glucose(FBG).Enzyme-linked immunosorbent assay(ELISA)was used to determine fasting insulin(FINS)and very-low-density-lipoprotein(VLDL)in rats,and to calculate the homeostasis model assessment of insulin resis⁃tance(HOMA-IR).Western blot was used to determine the expression levels of IRS-1,PI3K,p-PI3K,AKT,p-AKT,and mTORC1 proteins in liver tissues.Hematoxylin-eosin(HE)staining and oil red O staining were used to observe the morphological changes of liver tissues and the degree of hepatic steatosis in the rats of each group.Results:Compared with the normal group,the model group had significantly higher TC,TG,LDL-C,VLDL,FBG,FINS,and HOMA-IR,as well as expression of IRS-1,p-PI3K,p-AKT,and mTORC1 proteins in liver tissues,and a significantly lower HDL-C(all P<0.05);compared with the model group,the Qutan Huoxue decoction group had significant

关 键 词：非酒精性脂肪性肝病 祛痰活血方 胰岛素抵抗 IRS/PI3K/AKT/mTOR信号通路 

分 类 号：R89[医药卫生—法医学] R36[医药卫生—临床医学]