五倍子酸通过调节人肝癌HepG-2细胞线粒体氧化磷酸化对细胞凋亡的诱导作用  

Induction effect of gallic acid on apoptosis of human hepatocellular carcinoma HepG-2 cells through regulating mitochondrial oxidative phosphorylation

作　　者：张海英[1] 张文馨 赵文静[2] 李伟[1] ZHANG Haiying;ZHANG Wenxin;ZHAO Wenjing;LI Wei(Key Laboratory of Pathology,Ministry of Education,Jilin University,Changchun 130021,China;Central Laboratory,Hepatobiliary Disease Hospital of Jilin Province,Changchun 130062,China)

机构地区：[1]吉林大学基础医学院病理生物学教育部重点实验室,吉林长春130021 [2]吉林省肝胆病医院中心实验室,吉林长春130062 [3]吉林大学白求恩第二临床医学院

出　　处：《吉林大学学报（医学版）》2021年第1期125-132,共8页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科技发展计划自然科学基金项目(20170623093-06TC)。

摘　　要：目的:观察五倍子酸(GA)对人肝癌HepG-2细胞生长的抑制作用,并阐明其作用机制。方法:选择人肝癌HepG-2细胞,采用不同浓度(0,3.125、6.250、12.500、25.000、50.000和100.000 mg·L^-1)GA处理HepG-2细胞,分别作为对照组和3.125、6.250、12.500、25.000、50.000及100.000 mg·L^-1 GA组,采用MTT法检测各组细胞存活率。HepG-2细胞分为对照组及6.25,12.50,25.00 mg·L^-1 GA组,采用能量代谢分析仪检测线粒体氧化磷酸化(OXPHOS)动态全过程,流式细胞术检测各组细胞线粒体膜电位(MMP),比色法检测各组细胞中三磷酸腺苷(ATP)水平,Western blotting法检测各组细胞胞浆和线粒体中细胞色素C(Cyt C)蛋白表达量,吖啶橙染色观察各组细胞形态表现,AnnexinⅤ-FITC/PI双染流式细胞术检测各组细胞凋亡率。结果:与对照组比较,不同浓度GA组细胞存活率明显降低(P<0.05)。与对照组比较,6.25、12.50和25.00 mg·L^-1 GA组细胞基础有氧呼吸、最大有氧呼吸能力、有氧呼吸储备能力和ATP产生量均明显降低(P<0.05),细胞中MMP和ATP水平均明显降低(P<0.05),细胞胞浆中Cyt C蛋白表达量增加,线粒体中Cyt C蛋白表达量降低,吖啶橙染色可见细胞体积缩小、细胞核碎裂和染色质凝集,细胞早期凋亡率和晚期凋亡率明显升高(P<0.05)。结论:GA可以抑制HepG-2细胞增殖并诱导细胞凋亡,其机制可能与GA降低MMP和抑制线粒体OXPHOS功能有关。Objective:To observe the inhibitory effect of gallic acid(GA)on the growth of human hepatocellular carcinoma HepG-2 cells,and to elucidate its mechanism.Method:The human hepatocellular carcinoma HepG-2 cells were selected and treated with different concentrations(0,3.125,6.250,12.500,25.000,50.000,and 100.000 mg·L^-1)of GA and used as control group and 3.125,6.250,12.500,25.000,50.000,and 100.000 mg·L^-1 GA groups,respectively.The survival rates of cells in various groups were detected by MTT assay.The HepG2 cells were divided into control group and 6.25,12.50,and 25.00 mg·L^-1 GA groups.The mitochondrial oxidative phosphorylation(OXPHOS)of Hep-G2 cells was analyzed using energy metabolism analyzer.Flow cytometry was used to detect the mitochondrial membrane potential(MMP)of the cells in various groups.Colorimetry was used to detect the adenosine triphosphate(ATP)levels in the cells in various groups.The expression amounts of cytochrome C(Cyt C)protein in cytoplasm and mitochondria in various groups were detected by Western blotting method.The morphology of cells was measured by acridine orange staining,and AnnexinⅤ-FITC/PI double staining flow cytometry was used to detect the apoptotic rates of the cells in various groups.Results:Compared with control group,the survival rates of HepG2 cells in different concentrations of GA groups were significantly decreased(P<0.05).Compared with control group,the basal respiration,maximum respiration capacities,respiration reserve capacities and ATP levels in 6.25,12.50,and 25.00 mg·L^-1 GA groups were significantly decreased(P<0.05);the MMP and ATP levels in the cells were significantly reduced(P<0.05);the expression amounts of Cyt C protein in the cytoplasm were increased,and the expression amounts of Cyt C protein in mitochondria were decreased;the cells in GA groups were shrank,the nuclei were shattered,and chromatin was aggregated;the early apoptotic rates and late apoptotic rates were increased significantly(P<0.05).Conclusion:GA can inhibit the proliferation and i

关 键 词：五倍子酸 肝肿瘤 细胞凋亡 能量代谢 线粒体氧化磷酸化 

分 类 号：R285.5[医药卫生—中药学] R735.7[医药卫生—中医学]