吡格列酮通过AMPK/mTOR/SREBP-1通路抑制SREBP-1活性促进自噬改善代谢相关脂肪性肝病  

Pioglitazone inhibits SREBP-1 activity via AMPK/mTOR/SREBP-1 pathway and promotes autophagy to improve metabolic associated fatty liver disease

作　　者：廖星晨 田甜 张庆玉 吕昂 吴鹏波[1] 谭诗云[1] LIAO Xingchen;TIAN Tian;ZHANG Qingyu;LYU Ang;WU Pengbo;TAN Shiyun(Department of Gastroenterology,Renmin Hospital of Wuhan University/Hubei Key Laboratory of Digestive System Disease,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院消化内科/消化系统疾病湖北省重点实验室,湖北武汉430060

出　　处：《胃肠病学和肝病学杂志》2022年第10期1095-1100,共6页Chinese Journal of Gastroenterology and Hepatology

基　　金：湖北省自然科学基金(2018CFB236)。

摘　　要：目的探究吡格列酮(Pioglitazone,PGZ)通过AMPK/mTOR/SREBP-1通路对油酸(oleic acid,OA)诱导的代谢相关脂肪性肝病(metabolicassociated fatty liver disease,MAFLD)HepG2细胞模型SREBP-1蛋白表达、脂质蓄积、自噬及凋亡的影响。方法利用OA处理HepG2细胞24 h建造MAFLD细胞模型,分别用0(空白对照)、10、20、30、40、50μmol/L的PGZ处理HepG2细胞24 h,评估PGZ对细胞活力的影响。取对数生长期HepG2细胞分为对照组(CON组)、模型组(OA组)、PGZ低浓度组(PGZ-L组)、PGZ高浓度组(PGZ-H组),分别加入10μmol/L和30μmol/L PGZ干预其过程。用油红O染色法及Bodipy染色法观察各组细胞内脂滴蓄积情况,Western blotting法分别检测各组HepG2细胞p-AMPK、AMPK、p-mTOR、mTOR、SREBP-1、LC3、p62、Bax、Bcl-2蛋白表达水平的变化。结果CCK-8结果显示,在10~50μmol/L的浓度范围内,PGZ对HepG2细胞无明显细胞毒性。与CON组相比,OA组p-AMPK/AMPK、LC3-Ⅱ/Ⅰ比值、Bcl-2蛋白表达水平降低,p-mTOR/mTOR、SREBP-1、p62、Bax蛋白表达水平升高,自噬通量受损、凋亡水平升高。经过PGZ处理,这种情况被逆转,与OA组相比,PGZ-L组和PGZ-H组p-AMPK/AMPK、LC3-Ⅱ/Ⅰ比值、Bcl-2蛋白表达水平升高,p-mTOR/mTOR、SREBP-1、p62、Bax蛋白表达水平降低。与PGZ-L组比较,PGZ-H组上述指标的变化更为明显。结论PGZ可激活AMPK/mTOR/SREBP-1通路,抑制SREBP-1活性,促进自噬,抑制凋亡,改善OA诱导的MAFLD细胞模型内脂质蓄积。Objective To explore the effects of Pioglitazone(PGZ)on the lipid deposition,expression of SREBP-1 protein,autophagy and apoptosis via AMPK/mTOR/SREBP-1 pathway in HepG2 cell model of metabolic associated fatty liver disease(MAFLD)induced by oleic acid(OA).Methods HepG2 cells were treated with OA for 24 hours to build MAFLD cell models.HepG2 cells were treated with various concentrations of PGZ(0,10,20,30,40 and 50μmol/L)for 24 hours,to assess the effects of Pioglitazone on cell viability.The cells were divided into four groups:control group(CON group),model group(OA group),PGZ low-concentration group(PGZ-L group)and PGZ high-concentration group(PGZ-H group),and then 10μmol/L and 30μmol/L of PGZ were added to interfere with the process.The effect of PGZ on lipid deposition in HepG2 cells was detected by oil red O staining and Bodipy staining.Western blotting was used to detect the expression levels of p-AMPK,AMPK,p-mTOR,mTOR,SREBP-1,LC3,p62,Bax and Bcl-2 protein in HepG2 cells.Results The result of CCK-8 showed that PGZ didn′t reduce the viability of HepG2 cells in the range of 10μmol/L to 50μmol/L.Compared with the CON group,the expression levels of SREBP-1,Bax and p62 protein and the ratio of p-mTOR/mTOR in the OA group increased and the ratio of p-AMPK/AMPK and LC3-Ⅱ/Ⅰand the expression level of Bcl-2 protein decreased.The autophagy was inhibited and the apoptosis was induced.But the situation was reversed after interfering with PGZ.Compared with the OA group,the expression levels of SREBP-1,Bax and p62 protein and the ratio of p-mTOR/mTOR in the PGZ-L group and PGZ-H group decreased and the ratio of p-AMPK/AMPK and LC3-Ⅱ/Ⅰand the expression level of Bcl-2 protein increased.Compared with PGZ-L group,the changes of the above indicators in PGZ-H group were more obvious.Conclusion PGZ can activate AMPK/mTOR/SREBP-1 pathway to inhibit SREBP-1,promote autophagy,inhibit apoptosis,and improve lipid accumulation in MAFLD cell model induced by OA.

关 键 词：代谢相关脂肪性肝病 吡格列酮 甾醇调节元件结合蛋白1 雷帕霉素靶蛋白 腺苷酸活化蛋白激酶 自噬 

分 类 号：R575[医药卫生—消化系统]