摘要
目的探究抑制线粒体丙酮酸载体(mitochondrial pyruvate carrier,MPC)是否通过激活腺苷酸激活蛋白激酶(AMP-activated protein kinase,AMPK)-乙酰辅酶A羧化酶(acetyl CoA carboxylase,ACC)通路,改善棕榈酸(palmitic acid,PA)所致HepG2细胞脂肪变性情况。方法利用PA处理HepG2细胞作为高脂负荷模型组,实验分为正常对照组、模型组和MPC抑制组。油红O染色分析细胞的油脂蓄积情况。RT-PCR检测AMPK、ACC、CPT1A、SREBP1 mRNA表达水平。全自动蛋白分析仪检测AMPK、p-AMPK、ACC、p-ACC、SREBP1蛋白表达水平。结果PA可以诱导HepG2细胞胞质内聚集大量环状脂滴,降低p-AMPK、p-ACC蛋白表达水平,提高SREBP1基因以及蛋白表达水平。MPC抑制剂UK5099可以减少PA所致HepG2细胞胞质内脂滴数量,提高p-AMPK、p-ACC蛋白表达水平,降低SREBP1基因以及蛋白表达水平。结论体外抑制MPC可以明显改善PA所致HepG2细胞脂肪变性。激活AMPK-ACC信号通路,可能是MPC抑制剂防治非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)肝细胞脂肪变性的重要作用机制之一。
Aim To investigate the effect of inhibiting mitochondrial pyruvate carrier(MPC)by activating the AMP-activated protein kinase(AMPK)-acetyl-CoA carboxylase(ACC)signaling pathway on improving the lipid accumulation in HepG2 cells treated with palmitic acid(PA)and its mechanism.Methods The HepG2 cells were treated with PA to establish a steatosis model.The experiment included a control group,a model group and MPC inhibitor treatment group.Oil red O staining was used to examine the lipid accumulation.Changes in the mRNA expression of AMPK-ACC pathway related gene AMPK,ACC,carnitine palmitoyltransferase 1A(CPT1A),Sterol regulatory element-binding protein 1(SREBP1),which were related with lipid metabolism were assessed by RT-PCR.The expression of AMPK,p-AMPK,ACC,P-ACC,SREBP1 protein level was detected by automatic protein analyzer.Results PA significantly induced lipid accumulation,reduced the expression of p-AMPK,p-ACC protein level,and increased the expression of SREBP1 gene and protein level.MPC inhibitor UK5099 decreased the number of cell-containing lipid droplets induced by PA,increased the expression of p-AMPK and p-ACC protein level,and decreased the expression of SREBP1 gene and protein level.Conclusions In conclusion,inhibiting MPC can prevent the lipid accumulation of PA-induced HepG2 cells.To activate AMPK-ACC signaling pathway might be one of the most important mechanisms of MPC inhibitor’s prevention and treatment of non-alcoholic hepatic steatosis.
作者
李世英
汪艳
LI Shi-ying;WANG Yan(School of Pharmaceutical Science,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China;Central Lab,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China;Dept of Infectious Diseases and Liver Unit,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2020年第12期1679-1683,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81670522)
深圳市三名工程项目(No SZSM201911001)。
作者简介
李世英(1993-),女,硕士生,研究方向:非酒精性脂肪性肝病药理学,E-mail:1170930700@qq.com;通讯作者:汪艳(1973-),女,博士,教授,博士生导师,研究方向:慢性肝病基础与临床转化,E-mail:yanwang@smu.edu.cn。
