湖南中医药大学学报2025，Vol.45Issue(9)：1636-1643,8.DOI:10.3969/j.issn.1674-070X.2025.09.007
肝喜片通过干扰线粒体动力学诱导肝癌细胞HepG2凋亡的机制研究
Mechanism of Ganxi Tablet inducing apoptosis in hepatocellular carcinoma HepG2 cells by interfering with mitochondrial dynamics
摘要
Abstract
Objective To investigate the molecular mechanism by which Ganxi Tablet induces apoptosis in hepatocellular carcinoma HepG2 cells through regulating the mitochondrial dynamics mediated by mitofusin 2(MFN2)and dynamin-related protein 1(DRP1).Methods HepG2 cells were used as the model,with a blank control group and low-,medium-,and high-dose(10%,15%,and 20%drug-containing serum,respectively)Ganxi Tablet groups established.Real-time label-free cell analysis was employed to dynamically monitor cell proliferation.Flow cytometry was used to measure mitochondrial membrane potential,reactive oxygen species(ROS)levels,and the apoptosis rate.Western blot analysis was performed to assess the protein expression levels of MFN2,DRP1,poly(ADP-ribose)polymerase 1(PARP1),cleaved PARP1,Bcl-2-associated X protein(BAX),and cytochrome c(Cyt c).Results Compared with the blank control group,the cell index(CI)values of the medium-and high-dose Ganxi Tablet groups decreased(P<0.01);compared with the medium-dose Ganxi Tablet group,the CI value of the high-dose Ganxi Tablet group decreased(P<0.05).Compared with the blank control group,the proportion of cells with reduced mitochondrial membrane potential and intracellular ROS levels increased in all Ganxi Tablet groups(P<0.05,P<0.01);compared with the low-dose Ganxi Tablet group,the proportion of cells with reduced mitochondrial membrane potential increased in the medium-and high-dose Ganxi Tablet groups(P<0.05),and intracellular ROS levels increased in the high-dose Ganxi Tablet group(P<0.01).Compared with the blank control group,MFN2 protein expression level decreased in all Ganxi Tablet groups(P<0.05),while DRP1 protein expression level increased(P<0.05,P<0.01);compared with the medium-dose Ganxi Tablet group,MFN2 protein expression level decreased while DRP1 protein expression level increased in the high-dose Ganxi Tablet group(P<0.05).Compared with the blank control and low-dose Ganxi Tablet groups,the apoptosis rate increased in the medium-and high-dose Ganxi Tablet groups(P<0.01);compared with the medium-dose Ganxi Tablet group,the apoptosis rate increased in the high-dose Ganxi Tablet group(P<0.05).Compared with the blank control group,the cleaved PARP1/PARP1 ratio and BAX protein expression level increased in all Ganxi Tablet groups(P<0.05,P<0.01),while Cyt c protein expression level increased in the medium-and high-dose Ganxi Tablet groups(P<0.05,P<0.01).Compared with the low-dose Ganxi Tablet group,the cleaved PARP1/PARP1 ratio and Cyt c protein expression level increased in the medium-dose Ganxi Tablet group(P<0.05),and the cleaved PARP1/PARP1 ratio as well as BAX and Cyt c protein expression levels increased in the high-dose Ganxi Tablet group(P<0.05,P<0.01).Conclusion Ganxi Tablet suppresses MFN2 expression and activates DRP1-mediated mitochondrial fission,thereby disrupting mitochondrial dynamics.This disruption induces mitochondrial fragmentation,oxidative stress,and membrane potential collapse,subsequently promoting Cyt c release,BAX upregulation,and PARP1 cleavage,ultimately activating the mitochondrial-dependent apoptotic pathway.关键词
肝癌/线粒体动力学/HepG2/线粒体融合蛋白2/动力相关蛋白1/凋亡Key words
liver cancer/mitochondrial dynamics/HepG2/mitofusin 2/dynamin-related protein 1/apoptosis分类
医药卫生引用本文复制引用
曾千,吴涛,罗燕,罗吉..肝喜片通过干扰线粒体动力学诱导肝癌细胞HepG2凋亡的机制研究[J].湖南中医药大学学报,2025,45(9):1636-1643,8.基金项目
湖南省自然科学基金面上项目(2025JJ50499) （2025JJ50499）
湖南省自然科学基金医卫行业联合基金(2025JJ180792) （2025JJ180792）
湖南省中医药科研计划重点项目(A2024018) （A2024018）
湖南中医药大学研究生创新课题(2023CX0116). （2023CX0116）
