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目的探讨肿瘤坏死因子α(TNF-α)诱导PC12细胞凋亡以及对bax和bcl-xl基因表达的影响。方法以PC12细胞作为多巴胺神经元细胞模型,用MTT法测定不同浓度TNF-α对PC12细胞增殖的作用;用PI和AnnexinV进行细胞的双染凋亡检测及电镜检测PC12细胞凋亡;应用RT-PCR检测bax和bcl-xl基因mRNA表达的变化,用Westernblot检测bax和bcl-xl蛋白表达的变化。结果 TNF-α(1～100ng/ml)呈浓度依赖方式抑制PC12细胞增殖。30ng/mlTNF-α作用PC12细胞,PI双染及电镜均显示PC12细胞凋亡。RT-PCR表明药物作用后bcl-xlmRNA表达减少而baxmRNA表达增多;免疫印迹显示药物作用后bcl-xl蛋白表达减少而bax蛋白表达增多。结论 TNF-α诱导PC12细胞凋亡且呈量效关系,可能与bax和bcl-xl相关。
Objective To investigate the effects of tumor necrosis factor-α (TNF-α) on the apoptosis of PC12 cells and the gene expression of bax and bcl-xl. Methods PC12 cells were used as a model of dopamine neurons. MTT assay was used to determine the effect of different concentrations of TNF-α on the proliferation of PC12 cells. Double staining and Annexin V staining were used to detect the apoptosis of PC12 cells. The apoptosis of PC12 cells was detected by RT- PCR detection of bax and bcl-xl gene mRNA expression changes detected by Western blot bax and bcl-xl protein expression changes. Results TNF-α (1 ~ 100ng / ml) inhibited the proliferation of PC12 cells in a concentration-dependent manner. PC12 cells were treated with 30ng / ml TNF-α, PI double staining and electron microscopy showed PC12 cell apoptosis. The results of RT-PCR showed that the expression of bcl-xl mRNA decreased and the expression of bax mRNA increased after the drug treatment. Western blotting showed that the expression of bcl-xl protein and bax protein increased after the drug treatment. Conclusion TNF-α induces PC12 cell apoptosis in a dose-response manner, which may be related to bax and bcl-xl.