论文部分内容阅读
[目的]研究维生素B1(thiamine,VitB1)介导的Akt/mTOR/STAT3信号通路在谷氨酸(glutamate,Glu)诱导的PC12细胞损伤中的作用。[方法]应用MTT法检测不同浓度谷氨酸对PC12细胞作用不同时间的细胞生长抑制率,筛选出合适的作用浓度与作用时间后,将细胞分为4组,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+300μmol/L维生素B1处理组;D组:20 mmol/L谷氨酸+300μmol/L维生素B1+800nmol/L雷帕霉素(rapamycin,RAPA)处理组。应用流式细胞术观察各组作用12 h后细胞凋亡率,Western blot观察各组处理1、4、8、12 h后,p-Akt,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)A组的凋亡率为(3.42±0.79)%;C组凋亡率为(23.85±1.58)%,明显低于B组(40.20±2.54)%和D组(53.49±2.83)%(P<0.01);(3)Western blot检测结果表明C组各时间点p-Akt,p-mTOR,p-STAT3表达均高于A、B、D组,并且p-Akt表达量在1 h时最高,p-mTOR和p-STAT3在4 h时达到高峰。[结论]维生素B1激活了细胞Akt/mTOR/STAT3信号通路,该通路对谷氨酸诱导的神经细胞损伤起到了保护作用。
[Objective] To investigate the role of thiamine (VitB1) -mediated Akt / mTOR / STAT3 signaling pathway in the injury of PC12 cells induced by glutamate (Glu). [Methods] MTT assay was used to detect the cell growth inhibition rate of PC12 cells treated with different concentrations of glutamic acid. After screening the appropriate concentration and time, the cells were divided into 4 groups: A group: normal control group ; Group B: 20 mmol / L glutamic acid treatment group; Group C: 20 mmol / L glutamic acid + 300 μmol / L vitamin B1 treatment group; Group D: 20 mmol / L glutamic acid + 300 μmol / 800 nmol / L rapamycin (RAPA) treatment group. Flow cytometry was used to observe the apoptotic rate of each group for 12 h. The protein expressions of p-Akt, p-mTOR and p-STAT3 were detected by Western blot after treated for 1, 4, 8 and 12 h. [Results] (1) The inhibitory effect of glutamic acid on the growth of PC12 cells increased with the increase of time and concentration of action. (2) The apoptosis rate of group A was (3.42 ± 0.79)%. The apoptosis rate of group C was (23.85 ± 1.58)%, which was significantly lower than that of group B (40.20 ± 2.54)% and group D (53.49 ± 2.83)% respectively (P <0.01). (3) Western blot results showed that the expression of p-Akt, The expression of p-mTOR and p-STAT3 were higher than those in groups A, B and D, and the expression of p-Akt was the highest at 1 h. The p-mTOR and p-STAT3 peaked at 4 h. [Conclusion] Vitamin B1 activates the Akt / mTOR / STAT3 signaling pathway, which plays a protective role against glutamate-induced neuronal damage.