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目的探讨亲环素A(CyPA)对Aβ25-35诱导PC12细胞凋亡及氧化应激损伤的影响。方法先用不同浓度(0.1,1,10,100 nmol/L)的CyPA预处理PC12细胞30 min,再加入10μmol/L的Aβ25-35继续培养24 h或48 h,然后提取细胞进行试验。应用碘化丙啶(PI)单染,然后流式细胞仪检测细胞凋亡率,应用Western blot的方法检测caspase-3蛋白的表达,同时应用南京建成生物工程研究所的SOD试剂盒和GSH-PX试剂盒检测细胞内SOD和GSH-Px活性。结果1,10和100 nmol/L的rhCyPA预处理组与Aβ25-35单独孵育组相比,细胞的凋亡率明显下降,而caspase-3蛋白表达明显减少,10 nmol/L和100 nmol/L的CyPA可以增加细胞内SOD和GSH-Px的活性,而1 nmol/L的CyPA可以增加细胞内GSH-Px的活性。与Aβ25-35诱导组比较差异有统计学意义(P<0.05),且CyPA剂量越高效果越明显。结论 CyPA可以通过增加SOD和GSH-Px活性和减少caspase-3蛋白表达从而保护PC12细胞减少Aβ25-35诱导的细胞凋亡,且具有浓度依赖性。
Objective To investigate the effect of cyclophilin A (CyPA) on Aβ25-35-induced PC12 cell apoptosis and oxidative stress injury. Methods PC12 cells were pretreated with CyPA at different concentrations (0.1, 1, 10, and 100 nmol / L) for 30 min, followed by addition of 10 μmol / L Aβ25-35 for 24 h or 48 h. The cells were harvested and tested. The cells were stained with propidium iodide (PI), then the apoptosis rate was detected by flow cytometry. The expression of caspase-3 protein was detected by Western blot. At the same time, the SOD kit and GSH- PX kit to detect intracellular SOD and GSH-Px activity. Results The apoptotic rates of 1, 10 and 100 nmol / L rhCyPA pretreatment groups were significantly decreased compared with those of Aβ25-35 alone group, while the expression of caspase-3 protein was significantly decreased. The apoptosis rates of 10 nmol / L and 100 nmol / L CyPA can increase intracellular SOD and GSH-Px activity, while 1 nmol / L CyPA can increase intracellular GSH-Px activity. Compared with Aβ25-35 induction group, the difference was statistically significant (P <0.05), and the higher the dose of CyPA, the more obvious the effect. Conclusion CyPA protects PC12 cells from Aβ25-35-induced apoptosis by increasing the activity of SOD and GSH-Px and decreasing the expression of caspase-3 protein in a concentration-dependent manner.