论文部分内容阅读
目的通过已获得稳定表达葡萄糖调节蛋白(Grp75)的PC12细胞株,检测Grp75对缺糖诱导的细胞凋亡过程中Bax和NF-κB的影响。方法Grp75过表达组和对照组细胞无糖培养6h、12h2、4h和48h后,进行相关的实验。免疫印迹法检测缺糖状态下两组细胞中Grp75的表达水平和NF-κB的活性;应用半定量RT-PCR和免疫印迹法比较Bax表达水平的变化;免疫细胞化学通过构象特异性的Bax 6A7抗体检测Bax的活化。结果Bax活化和NF-κB活性的下降在缺糖诱导的PC12细胞凋亡过程中发挥了重要的作用。而Grp75通过阻止Bax的活化和NF-κB活性的下降抑制缺糖诱导的凋亡。缺糖状态下Grp75过表达组和对照组细胞中Bax表达水平均未发生改变。结论Bax活化和NF-κB活性下降与缺糖诱导的PC12细胞凋亡关系密切,Grp75通过阻止Bax的活化和维持NF-κB的活性保护PC12细胞。
OBJECTIVE: To investigate the effect of Grp75 on Bax and NF-κB during glucose deprivation-induced apoptosis in PC12 cell line, which has been stably expressed glucose regulatory protein (Grp75). Methods Grp75 overexpression group and control group of cells without sugar culture 6h, 12h2, 4h and 48h after the relevant experiments. The expression of Grp75 and the activity of NF-κB in two groups of cells were detected by Western blotting. The changes of Bax expression were compared by semi-quantitative RT-PCR and Western blotting. Immunocytochemistry was performed by conformation-specific Bax 6A7 Antibodies detect activation of Bax. Results The decrease of Bax activation and NF-κB activity played an important role in the process of glucose-induced PC12 cell apoptosis. While Grp75 inhibits glucose-induced apoptosis by preventing the activation of Bax and the decrease of NF-κB activity. No expression of Bax in Grp75 overexpression group and control group was observed under the condition of glucose deprivation. Conclusion The decrease of Bax activation and NF-κB activity are closely related to the apoptosis of PC12 cells induced by glucose deprivation. Grp75 protects PC12 cells by inhibiting the activation of Bax and maintaining the activity of NF-κB.