北京中医药大学学报2025，Vol.48Issue(2)：193-204,12.DOI:10.3969/j.issn.1006-2157.2025.02.008
截断逆挽方调控AMPK改善HepG2细胞线粒体损伤的机制
Jieduan Niwan Formula regulates AMPK to alleviate mitochondrial damage in HepG2 cells
摘要
Abstract
Objective To investigate the regulatory effect of Jieduan Niwan Formula(JDNWF)drug-containing serum on AMPK-mediated mitochondrial quality control in D-GalN-induced HepG2 cells.Methods Twenty male Wistar rats were randomly divided into blank control and JDNWF-containing serum groups,10 rats per group.The JDNWF-containing serum group was gavaged with JDNWF(21.7 g/kg),whereas the blank control group was gavaged with saline.Blood was collected to prepare JDNWF-containing and blank control serum.Cell viability,mitochondrial damage indicators,and MQC pathway protein expression levels were evaluated to determine the optimal volume fraction of JDNWF.HepG2 cells were divided into control,D-GalN,DMSO,AMPK inhibitor,JDNWF drug-containing serum,and JDNWF drug-containing serum plus AMPK inhibitor groups,and corresponding drug interventions were administered to each group.Cells were collected after the interventions,and the CCK-8 assay was used to measure cell viability,the 2'-7'-dichlorodihydrofluorescein diacetate fluorescent probe was used to detect reactive oxygen species(ROS)levels,JC-1 was used to detect mitochondrial membrane potential,thiobarbituric acid was used to measure malondialdehyde(MDA)levels,WST-8 was used to measure superoxide dismutase(SOD)activity,and western blotting was used to detect the expression levels of mitochondrial quality control-related proteins,including p-AMPK,AMPK,PGC-1α,NRF1,TFAM,MFN2,and DRP1.Results 5%JDNWF drug-containing serum most significantly restored cell viability,mitochondrial damage markers,and MQC pathway protein expression in the model group.Therefore,it was chosen for intervention in subsequent experiments.Compared to the control group,the cell viability of the D-GalN,DMSO,and AMPK inhibitor groups was significantly reduced(P<0.01).In contrast,the heterogeneity of mitochondrial membrane potential,ROS,and MDA levels was significantly increased(P<0.01),and SOD activity was significantly decreased(P<0.01).The p-AMPK,PGC-1α,NRF1,TFAM,MFN2,and DRP1 protein expression levels were significantly decreased(P<0.01).After JDNWF drug-containing serum intervention,compared to the DMSO group,cell viability significantly increased(P<0.01),mitochondrial membrane potential heterogeneity,ROS,and MDA levels significantly decreased(P<0.01),SOD activity significantly increased(P<0.01),and p-AMPK,PGC-1α,NRF1,TFAM,and MFN2 protein expression levels significantly increased(P<0.01),whereas DRP1 protein expression significantly decreased(P<0.01).Compared to the JDNWF drug-containing serum group,the cell viability in the JDNWF plus AMPK inhibitor group significantly decreased(P<0.01),mitochondrial membrane potential heterogeneity and ROS levels significantly increased(P<0.01),MDA levels significantly increased(P<0.05),SOD activity significantly decreased(P<0.05),p-AMPK,PGC-1α,NRF1,and TFAM protein expression levels significantly decreased(P<0.01),MFN2 protein expression significantly decreased(P<0.05),and DRP1 protein expression significantly increased(P<0.01).Conclusion JDNWF drug-containing serum may restore mitochondrial function and improve D-GalN-induced HepG2 cell injury by regulating AMPK-mediated mitochondrial quality control.关键词
截断逆挽方/肝细胞损伤/线粒体质量控制/单磷酸腺苷激活的蛋白激酶/慢加急性肝衰竭Key words
Jieduan Niwan Formula/hepatocyte injury/mitochondrial quality control/amp-activated protein kinase/acute-on-chronic liver failure分类
医药卫生引用本文复制引用
马瑞旻,王瀚婧,张文馨,马重阳,张秋云,杜宇琼..截断逆挽方调控AMPK改善HepG2细胞线粒体损伤的机制[J].北京中医药大学学报,2025,48(2):193-204,12.基金项目
国家自然科学基金项目(No.82074237,No.81573767) （No.82074237,No.81573767）
北京市中医药科技发展基金项目(No.JJ-2020-51) National Natural Science Foundation of China(Nos.82074237 and 81573767) （No.JJ-2020-51）
