论文部分内容阅读
目的:研究谷氨酸(glutamate,Glu)诱导PC12细胞损伤后mTOR/STAT3信号通路的表达情况及对细胞损伤的保护作用。方法:用不同浓度谷氨酸作用不同时间诱导PC12细胞损伤,筛选出合适的浓度和作用时间后,将细胞分为3组进行下一步实验,分别为A组:正常对照组;B组:20 mmol/L谷氨酸处理组;C组:20 mmol/L谷氨酸+800 nmol/L雷帕霉素(rapamycin,RAPA)处理组。应用流式细胞术检测各组处理12 h后细胞凋亡率,Western blot观察各组处理1 h、4 h、8 h、12 h后,p-mTOR,p-STAT3蛋白表达情况。结果:(1)谷氨酸对PC12细胞的生长抑制作用随作用时间和作用浓度的增加而增强。(2)C组凋亡率明显高于A组和B组。(3)Western blot检测结果表明B组各时间点p-mTOR,p-STAT3表达均高于A、C组,并在4 h时达到高峰。结论:细胞损伤激活了mTOR/STAT3信号通路,该通路的激活减少了细胞凋亡,对谷氨酸导致的神经细胞损伤具有保护作用,有助于神经损伤的修复。
AIM: To investigate the expression of mTOR / STAT3 signaling pathway and its protective effect on cell injury induced by glutamate (Glu) in PC12 cells. Methods: The injury of PC12 cells was induced by different concentrations of glutamic acid for different periods of time, and the appropriate concentration and time were screened. The cells were divided into 3 groups for further experiments: group A: normal control group; group B: 20 mmol / L glutamate; Group C: 20 mmol / L glutamate + 800 nmol / L rapamycin (RAPA). Flow cytometry was used to detect the apoptosis rate of each group after treatment for 12 h. The expression of p-mTOR and p-STAT3 protein in each group were detected by Western blot at 1 h, 4 h, 8 h and 12 h after treatment. Results: (1) The growth inhibitory effect of glutamic acid on PC12 cells increased with time and concentration. (2) The apoptosis rate in group C was significantly higher than that in group A and group B. (3) The results of Western blot showed that the expression of p-mTOR and p-STAT3 in group B at each time point were higher than those in group A and C, and peaked at 4 h. Conclusion: The cell injury activates the mTOR / STAT3 signaling pathway. Activation of this pathway reduces apoptosis and protects neurons against glutamate-induced neuronal injury, contributing to the repair of nerve injury.