柴胡皂苷d对抗线粒体损伤保护人肝细胞作用研究

Study on Protective Effect of Bupleurin D against Mitochondrial Injury in Hepatocytes

目的观察柴胡皂苷d(SSd)对抗线粒体损伤保护人肝细胞的作用,探讨SSd抗氧化应激作用的机制。方法体外培养人正常肝细胞L02,采用H_(2)O_(2)诱导L02细胞氧化应激损伤模型,干预组加入中药单体SSd孵育24 h后再经过H_(2)O_(2)处理细胞,CCK8法检测SSd干预L02细胞损伤后细胞活力,试剂盒检测ALT、AST、MDA水平,荧光染色法检测线粒体膜电位(JC-1)、线粒体超氧化物(MitoSox Red)、化学发光发检测ATP水平。结果SSd能抑制L02细胞损伤引起的细胞活力下降,降低氧化应激损伤L02细胞的ALT、AST、MDA水平;抑制JC-1损伤及ATP水平下降,降低MitoSox Red水平。结论SSd对氧化应激损伤的L02细胞有保护作用,与维持线粒体正常功能、降低活性氧水平相关。

Objective To observe the effect of bupleuronin d(SSd)on protecting hepatocytes from mitochondrial injury and explore the mechanism of SSd anti-oxidative stress.Methods In vitro cultivation of L02 human normal liver cells,H_(2)O_(2) was used to induce L02 cell oxidative stress damage model.The intervention group added Chinese medicine monomer SSd again after incubation for 24 h after H_(2)O_(2) treatment cells.CCK8 method was ued to detect the SSd intervention L02 cell damage after the cell vitality.Levels of ALT,AST and MDA were detected by kits.The fluorescent staining method was used to detect JC-1 and MitoSox Red.The ATP level was detected by chemil uminescence method.Results SSd could inhibit the decrease of cell activity caused by L02 cell injury,reduce levels of ALT,AST and MDA in L02 cells damaged by oxidative stress and inhibit JC-1 damage and ATP level decline,and MitoSox Red level was decreased.Conclusion SSd has a protective effect on L02 cells damaged by oxidative stress,which is related to maintaining normal mitochondrial function and reducing reactive oxygen species.