中国中药杂志2025，Vol.50Issue(18)：5045-5054,10.DOI:10.19540/j.cnki.cjcmm.20250409.706
基于脂噬探讨雷公藤甲素引起肝细胞脂毒性的机制研究
Mechanism of triptolide-induced hepatic cell lipotoxicity based on lipophagy
摘要
Abstract
This study aimed to investigate the effects of triptolide on lipophagy and to explore the role and underlying mechanism of lipophagy in triptolide-induced hepatocellular injury.HL7702 human normal hepatocytes and C57BL/6J mice were treated with 25,50,or 100 nmol·L-1 or 0.4,0.6,or 0.8 mg·kg-1 triptolide,respectively.Additionally,HL7702 cells were co-treated with Ras-related protein 7(Rab7)small interfering RNA(siRNA)and 50 nmol·L-1 triptolide.Cell viability was assessed by calcein AM/PI double staining,and liver histopathological changes were observed using hematoxylin-eosin(HE)staining.Lipid droplet content was examined via oil red O and nile red staining.Commercial kits were used to measure the content of triglycerides(TG),total cholesterol(TC),free fatty acids(FFAs),glutathione(GSH),and malondialdehyde(MDA),the activities of alanine aminotransferase(ALT)and aspartate aminotransferase(AST),and the levels of mitochondrial membrane potential(MMP)and intracellular reactive oxygen species(ROS).The expression of microtubule-associated protein 1 light chain 3 Ⅱ(LC3 Ⅱ),selective autophagy adaptor p62,Rab7,and perilipin 2(Plin2)were detected by qRT-PCR or Western blot.The results showed that in vitro,25,50 and 100 nmol·L-1 triptolide significantly reduced the cell survival rate,upregulated the expression of Rab7 and LC3 Ⅱ,downregulated the expression of p62 and Plin2,reduced the content of TG,TC,GSH and MMP,and increased the content of FFAs,MDA and ROS.In vivo,0.4,0.6 and 0.8 mg·kg-1 triptolide increased the activity of ALT and AST in serum,caused liver tissue damage,upregulated the expression of Rab7 and LC3 Ⅱ,downregulated the expression of p62,decreased the content of TG,TC and GSH,and increased the content of FFAs and MDA.Rab7 siRNA significantly increased the levels of TG,TC and MMP,decreased the levels of FFAs and ROS,and increased the cell survival rate.In conclusion,triptolide at certain concentrations may induce Rab7-mediated lipophagy to promote lipid droplet degradation,thereby causing hepatocellular lipotoxicity,leading to mitochondrial dysfunction and oxidative stress.关键词
雷公藤甲素/脂噬/肝细胞毒性/线粒体功能损伤/氧化应激/脂毒性Key words
triptolide/lipophagy/hepatotoxicity/mitochondrial dysfunction/oxidative stress/lipotoxicity引用本文复制引用
刘珊,苏越蕊,李宏岩,周琨,韩波,任晋宏,李鹏程,魏砚明..基于脂噬探讨雷公藤甲素引起肝细胞脂毒性的机制研究[J].中国中药杂志,2025,50(18):5045-5054,10.基金项目
山西省科技厅基础研究课题(202103021224295,202303021222294) （202103021224295,202303021222294）
山西省回国留学人员科研资助项目(2022-166,2024-134) （2022-166,2024-134）
基于炎性反应的重大疾病创新药物山西省重点实验室开放基金项目(SXInFDL2024-009) （SXInFDL2024-009）
