肉苁蓉总苷对原代培养小鼠肝细胞酒精损伤保护作用研究

Protective effect of cistanchis glycosides on ethanol-induced damage in the primary hepatocytes of mice

目的:研究肉苁蓉总苷对原代培养肝细胞的保护作用。方法:采用原位灌流法收集原代肝细胞,MTT法评价肉苁蓉总苷对酒精损伤原代培养肝细胞存活率的影响;荧光显微镜技术评价肉苁蓉总苷对酒精损伤原代培养肝细胞及细胞核形态的影响;流式细胞仪检测肉苁蓉总苷对酒精损伤原代培养肝细胞凋亡的影响;免疫细胞化学染色法检测肉苁蓉总苷对bcl-2和c-fos表达的影响。结果:原代肝细胞经酒精损伤后,存活率降低,出现明显凋亡和坏死改变,凋亡抑制基因bcl-2减弱、促凋亡基因c-fos表达增强。肉苁蓉总苷可明显提高细胞存活率,改善凋亡和坏死情况,增强bcl-2表达,抑制c-fos表达。且作用呈剂量依赖性。结论:肉苁蓉总苷可通过增加凋亡抑制基因bcl-2表达,减少促凋亡基因c-fos表达,减少凋亡和坏死,增加细胞存活率来实现对原代培养肝细胞的保护作用。

AIM： To discuss the protection of cistanchis glycosides on ethanol-induced damage in the primary hepatocyte of mice. METHODS： The primary hepatocytes were collected by perfusion in situ method.The cell livability was evaluated by MTT,fluorescence microscopy and flow cytometry were used to evaluate the cell apoptosis and necrosis,immunohistochemistry was used to detect the expression of bcl-2 and c-fos.RESULTS：After the primary hepatocytes were damaged by alcohol,the cell livability decreased,apoptosis and necrosis were obvious,the expression of bcl-2 was decreased while that of c-fos was increased.Cistanchis glycosides could increase the cell livability,ameliorate apoptosis and necrosis,increase the expression of bcl-2,and decrease the expression of c-fos in a dose-dependent manner.CONCLUSION： The protective effects of cistanchis glycosides on ethanol-induced liver damage in the primary hepatocytes is associated with increasing the expression of bcl-2,and decreasing the expression of c-fos;resulting less apoptosis,less necrosis,and increasing the cell livability.