牛黄参含药血清诱导人肝癌细胞HepG-2凋亡的机理研究

Serum Containing Niuhuanshen Capsule Induces Apoptosis of Human Hepatocellular Carcinoma HepG2 Cell

目的探讨牛黄参胶囊含药血清对人肝癌Hep G-2细胞凋亡及Fas表达的影响。方法将实验分为对照组,环磷酰胺组,牛黄参胶囊含药血清5%、10%和20%组。采用DNA Ladder观察DNA凋亡条带的形成,运用荧光显微镜观察凋亡细胞核的形态,采用流式细胞术检测其诱导细胞凋亡的凋亡率;分别采用免疫组织化学和流式细胞术检测牛黄参胶囊含药血清对Hep G2细胞表面Fas表达的影响。结果牛黄参胶囊含药血清作用于Hep G2细胞48 h后,有明显的DNA凋亡条带的形成,Hep G2细胞核染色质明显固缩和碎裂,产生凋亡小体;其促进Hep G2细胞的凋亡具有明显的浓度依赖性(P<0.01);能明显上调Hep G2细胞表面Fas的表达,具有明显的剂量依赖性(P<0.01)。结论牛黄参胶囊含药血清能促进人肝癌Hep G-2细胞凋亡,上调Fas蛋白表达可能是其抗肝癌药效机制之一。

Objective To investigate the effects of the serum containing Niuhuangshen capsule( NHS) on the apoptosis of human hepatoma carcinoma Hep G2 cell and on the expression Fas in Hep G2. Methods The effects of the NHS on apoptosis of human hepatoma carcinoma Hep G2 were determined by DNA ladder and fluorescence microscope. The cell apoptosis percentage was examined by using flowcytometry. Immunohistochemistry and flowcytometry were used to detect the expression of Fas in the level of protein. Results In 48 h,NHS inuced the cell shrinkage,cell pyknosis,chromosomes condensation and DNA fragment. In addition,the flow cytometric analysis revealed NHS( 5%,10%,20%) dose- dependently incresed the apoptosis percentage of Hep G2 cell. Immunohistochemistry and flowcytometry demonstrated the expression of Fas was increased with dose dependent. Conclusion The serum containing Niuhuangshen capsule probably induced Hep G2 apoptosis by taking advantage of up- regulation of Fas.