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目的探讨缺血后处理对PC12细胞缺血再灌注损伤引起的细胞凋亡的作用及其作用机制。方法将PC12细胞分为3组:正常组、缺血再灌注组、缺血后处理组。缺血再灌注组予以糖氧剥夺12 h后正常培养,缺血后处理组经糖氧剥夺12 h后予以3个循环的正常培养(10 min)→糖氧剥夺(10 min),再正常培养12 h后通过Hoechst染色检测各组细胞的凋亡情况,应用Westernblot检测各组细胞Caspase-3活化蛋白及磷酸化NF-κB/p65蛋白表达水平,采用RT-PCR检测各组细胞NF-κB及Caspase-3 mRNA表达水平。结果Hoechst染色显示缺血后处理可降低缺血再灌注引起的细胞凋亡;与对照组相比,缺血再灌注组磷酸化NF-κB/p65和Cleaved caspase-3的蛋白表达水平高;缺血后处理组磷酸化NF-κB/p65和Cleaved caspase-3的蛋白表达水平明显低于缺血再灌注组;NF-κB和Caspase-3的mRNA表达趋势与蛋白表达基本一致。结论缺血后处理可以减轻缺血再灌注损伤引起的PC12细胞凋亡,这可能与NF-κB/p65信号通路有关。
Objective To investigate the effect and mechanism of ischemic postconditioning on apoptosis induced by ischemia-reperfusion injury in PC12 cells. Methods PC12 cells were divided into 3 groups: normal group, ischemia-reperfusion group and ischemic postconditioning group. The rats in ischemia-reperfusion group were cultured for 12 h after oxygen-glucose deprivation, and the rats in normal control group were subjected to 3 cycles of normal culture (10 min) and glucose-oxygen deprivation (10 min) After 12 h, the apoptosis of cells in each group was detected by Hoechst staining. The expressions of Caspase-3 activation protein and phosphorylated NF-κB / p65 protein were detected by Western blot. The expressions of NF-κB and NF- Caspase-3 mRNA expression level. Results Hoechst staining showed that ischemic postconditioning could reduce the apoptosis induced by ischemia-reperfusion. Compared with the control group, the protein expression of phosphorylated NF-κB / p65 and Cleaved caspase-3 in ischemia-reperfusion group was high; The protein expression of phosphorylated NF-κB / p65 and Cleaved caspase-3 in the post-hematopoietic group was significantly lower than that in the ischemia-reperfusion group. The mRNA expression trends of NF-κB and Caspase-3 were basically consistent with the protein expression. Conclusions Ischemia treatment can reduce ischemia-reperfusion injury-induced apoptosis of PC12 cells, which may be related to NF-κB / p65 signaling pathway.