护肝片对非酒精性脂肪性肝病小鼠细胞自噬、焦亡和PI3K/Akt/mTOR信号通路的影响  

Effects of Hugan Tablets on autophagy,pyroptosis and PI3K/Akt/mTOR signaling pathway in a mouse model of non-alcoholic fatty liver disease

作　　者：王瑞华 黄兰蔚 徐列明[1,3] 平键[1,3] WANG Rui-hua;HUANG Lan-wei;XU Lie-ming;PING Jian(Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai Baoshan Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai 201999,China;Shanghai Key Laboratory of Traditional Chinese Medicine,Ministry of Education Key Laboratory of Liver and Kidney Diseases,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院,上海201203 [2]上海市宝山区中西医结合医院,上海201999 [3]上海中医药大学肝肾疾病病证教育部重点实验室,上海市中医临床重点实验室,上海201203

出　　处：《中成药》2025年第3期766-773,共8页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金青年基金项目(81773980)。

摘　　要：目的探讨护肝片对高脂饮食诱导的非酒精性脂肪性肝病(NAFLD)小鼠的干预作用及对自噬、焦亡和PI3K/Akt/mTOR信号通路的影响。方法C57BL/6小鼠随机分为正常组、模型组、护肝片组(0.7 g/kg)和易善复组(0.23 g/kg),每组10只,采用高脂饮食喂养16周诱导NAFLD模型,第13周起,灌胃给予相应剂量药物,16周末处死小鼠,采集血清及肝脏组织样本。检测血清ALT、AST、TG、TC、LDL水平,肝组织TG、TC、NEFA、MDA水平和SOD、GSH-Px活性;ELISA法检测血清IL-1β、IL-6、TNF-α水平;HE染色及油红O染色观察肝组织病理变化;Western blot法检测肝组织ACC、CPT1A、FAS、p-PI3K、p-Akt、p-mTOR、P62、LC3、NLRP3、GSDMD、Caspase1蛋白表达。结果与模型组比较,护肝片组小鼠体质量、肝体比降低(P<0.01);血清ALT、AST、TG、TC、LDL、IL-6、IL-1β、TNF-α水平降低(P<0.05,P<0.01);肝组织TG、TC、NEFA、MDA水平升高(P<0.05),SOD、GSH-Px活性降低(P<0.05);肝组织中脂质沉积、气球样变等病理改变有所改善,NAS评分和油红O染色面积均降低(P<0.01);肝组织ACC1、FAS、NLRP3、Caspase1、GSDMD、P62、p-PI3K、p-Akt、p-mTOR蛋白表达降低(P<0.05,P<0.01),CPT1A、LC3蛋白表达升高(P<0.01)。结论护肝片可有效防治高脂饮食诱导小鼠NAFLD的形成,其机制可能与抑制PI3K/Akt/mTOR信号通路,促进肝细胞自噬,抑制细胞焦亡有关。AIM To observe the effects of Hugan Tablets on high-fat diet induced non-alcoholic fatty liver disease(NAFLD)in a mouse model,and the autophagy,pyroptosis and the PI3K/Akt/mTOR signaling pathway as well.METHODS The C57BL/6 mice were randomly divided into the normal group,the model group,the Hugan Tablets group(0.7 g/kg)and the Yishanfu group(0.23 g/kg),with 10 mice in each group.The NAFLD mouse model was established by 16 weeks feeding of high-fat diet.From the 13th week,the mice started their corresponding dosing of the drug by gavage followed by killing of the mice at the end of 16th week and collection of their serum and liver tissue samples.The mice had their serum ALT,AST,TG,TC,LDL levels,liver TG,TC,NEFA,MDA levels and activities of SOD and GSH-Px detected;their serum levels of IL-1β,IL-6 and TNF-αdetected by ELISA;their hepatic pathological changes observed using HE staining and oil red O staining;and their hepatic protein expressions of ACC,CPT1A,FAS,p-PI3K,p-Akt,p-mTOR,P62,LC3,NLRP3,GSDMD and Caspase1 detected by Western blot.RESULTS Compared with the model group,the Hugan Tablets group displayed decreased body weight and hepatosmatic index level(P<0.01);decreased levels of serum ALT,AST,TG,TC,LDL,IL-6,IL-1βand TNF-α(P<0.05,P<0.01);increased hepatic levels of TG,TC,NEFA and MDA(P<0.05);decreased activities of SOD and GSH-Px(P<0.05);improved pathological changes of hepatic lipid deposition and hepatocytic ballooning and decreased NAS score and oil red O staining area(P<0.01);decreased hepatic protein expressions of ACC1,FAS,NLRP3,Caspase1,GSDMD,P62,p-PI3K,p-Akt and p-mTOR(P<0.05,P<0.01);and increased protein expressions of CPT1A and LC3(P<0.01).CONCLUSION Hugan Tablets can effectively prevent and control the development of high-fat diet induced NAFLD in mice,and the mechanism may be associated with the promotion of autophagy in hepatocytes and the inhibition of pyroptosis via the inhibition of PI3K/Akt/mTOR signaling pathway.

关 键 词：护肝片 非酒精性脂肪性肝病 自噬 焦亡 PI3K/Akt/mTOR信号通路 

分 类 号：R285.5[医药卫生—中药学]