摘要
[目的]探讨人参皂苷Re对乙醇诱导的大鼠H4IIE肝细胞线粒体功能损伤的作用机制.[方法]采用MTT法检测不同浓度乙醇分别刺激H4IIE细胞不同时间时的细胞活力水平,确定建立模型的乙醇浓度及作用时间.实验分为空白对照组、乙醇(EtOH 800 mmol/L)模型组、人参皂苷Re(10μmol/L)组和人参皂苷Re+乙醇(Re 10μmol/L+EtOH 800 mmol/L)组.采用流式细胞仪及荧光显微镜分别检测各组细胞活性氧(ROS)和膜电位(MMP)的变化情况,并使用多功能酶标仪检测各组ATP水平变化;采用Western blot法检测各组氧化应激相关蛋白Keap1、Nrf2、HO-1及Bach1蛋白表达水平.[结果]MTT法检测结果显示,与空白对照组比较,随着乙醇浓度的升高及作用时间的延长,各组细胞存活率明显降低(P<0.05).与空白对照组比较,乙醇模型组ROS生成明显增多,且MMP、ATP水平明显降低,差异均具有统计学意义(P<0.05);乙醇模型组Keap1、total-Nrf2、nucleus-Nrf2、HO-1及Bach1表达均明显升高(P<0.05).[结论]乙醇可降低大鼠H4IIE肝细胞生存率,且在一定范围内呈时间-剂量效应关系;乙醇可致大鼠H4IIE肝细胞线粒体功能损伤;人参皂苷Re对酒精性肝细胞损伤具有一定的保护作用,其机制认为可能与保护线粒体功能及Nrf2-ARE通路的抗氧化作用有关系.
OBJECTIVE To investigate the mechanism of ginsenoside Re on mitochondrial function injury induced by alcohol in rat H4IIE hepatocytes.METHODS MTT method was used to detect the cell viability of H4IIE cells stimulated by different concentrations of ethanol at different times,and to determine the ethanol concentration and action time of the model.The experiment was divided into blank control group,ethano(l EtOH 800 mmol/L)model group,ginsenoside Re(Re 10μmol/L)group,and ginsenoside Re+ethanol(Re 10μmol/L+EtOH 800 mmol/L)group.The changes of reactive oxygen species(ROS)and membrane potential(MMP)were detected by flow cytometry and fluorescence microscope,and the changes of ATP levels in each group were detected by multifunctional microplate reader.The expressions of oxidative stress related proteins Keap1,Nrf2,HO-1 and Bach1 were detected by Western blot.RESULTS MTT assay showed that compared with the blank control group the cell survival rate of each group was significantly decreased with the increase of ethanol concentration and extension of action time(P<0.05).Compared with the blank control group,ROS production was significantly increased,MMP and ATP levels were significantly decreased in the ethanol model group,with statistical significance(P<0.05).The expressions of Keap1,total-Nrf2,nucleus-Nrf2,HO-1 and Bach1 in the ethanol model group were significantly increased(P<0.05).CONCLUSION Ethanol can reduce survival rate of rat H4IIE hepatocytes with a time dose effect relationship in a certain range.Ethanol can damage mitochondrial function of rat H4IIE hepatocytes.Ginsenoside Re has a certain protective effect on alcoholic hepatocyte injury,and its mechanism may be related to the protection of mitochondrial function and the antioxidant effect of Nrf2-ARE pathway.
作者
尹春金
李先燕
金贵虹
权修权
YIN Chunjin;LI Xianyan;JIN Guihong;QUAN Xiuquan(Department of Respiratory and Critical Care Medicine,Dunhua Hospital,Dunhua 133700,Jilin,China;Department of Emergency Medicine,Affiliated Hospital of Yanbian University,Yanji 133000,Jilin,China)
出处
《延边大学医学学报》
CAS
2023年第4期241-248,共8页
Journal of Medical Science Yanbian University
