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目的:研究胰岛素抗MPP+诱导PC12细胞凋亡中胰岛素受体(IR)的变化。方法:应用半定量逆转录聚合酶链式反应(RT-PCR)测定胰岛素抗MPP+诱导PC12细胞凋亡过程中IRmRNA的基因表达,免疫沉淀Western印迹分析技术测定相同条件下IR的蛋白表达,应用IR自身磷酸化的特异阻断剂HNMPA-AM3,通过MTT法观察PC12细胞生存率的改变、瞬转IR质粒后再次观察细胞生存率的改变及进一步通过免疫沉淀Western印迹分析技术检测IR蛋白质磷酸化水平。结果:IR磷酸化程度的变化与细胞生存率的变化有关,HNMPA-AM3可阻断胰岛素对MPP+诱导的PC12细胞的抗凋亡作用,IR可增强胰岛素的抗凋亡作用。结论:IR磷酸化增加可抵抗MPP+诱导的PC12细胞的凋亡。
AIM: To investigate the changes of insulin receptor (IR) in PC12 cells induced by anti-MPP +. Methods: The gene expression of IRmRNA in PC12 cells induced by insulin against MPP + was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), and the protein expression of IR was detected by immunoprecipitation Western blot analysis. IR HNMPA-AM3, a specific inhibitor of autophosphorylation, was used to observe the changes of survival rate of PC12 cells by MTT method. The changes of cell survival rate were observed again after transfection of IR plasmid and further the phosphorylation level of IR protein was detected by immunoprecipitation Western blot analysis . Results: The change of IR phosphorylation was related to the change of cell survival rate. HNMPA-AM3 could block the anti-apoptotic effect of insulin on MPP + -induced PC12 cells, and IR could enhance the anti-apoptotic effect of insulin. Conclusion: The increase of IR phosphorylation can resist the apoptosis of PC12 cells induced by MPP +.