甲基苯丙胺对PC12细胞中二硫键异构酶S亚硝基化的影响

Effect of methamphetamine exposure on S-nitrosylation of protein disulphide isomerase in PC12 cells

目的研究甲基苯丙胺(METH)对PC12细胞中二硫键异构酶(PDI)S亚硝基化的影响以及对神经元的毒性作用。方法不同浓度的METH处理PC12细胞,CCK-8法检测细胞存活率,L-NNA与不同浓度的METH共同处理PC12细胞,生物素转化法检测各细胞内PDI及S亚硝基化PDI(PDI-SNO)表达情况,CCK-8法检测细胞存活率情况,HE染色法观察细胞形态学变化。结果METH使得PC12细胞存活率降低,PDI-SNO表达率增高,当同时用一氧化氮合酶抑制剂N-硝基-L-精氨酸(L-NNA)与METH共同处理细胞时,L-NNA可有效地提高细胞的存活率,抑制PDI-SNO的表达率。结论 METH可对PC12细胞产生明显毒性作用,且使PC12细胞中的PDI发生显著的S亚硝基化。

Objective To study the effect of methamphetamine （METH） exposure on S-nitrosylation of protein disulphide isomerase and the neurotoxicity of METH in PC12 cells. Methods PC12 cells were exposed to different concentrations of METH, and the cell viability was assessed using the cell-counting kit-8. PC12 cells exposed to METH in the presence of the NOS inhibitor N-nitro-L-arginine （L-NNA） were examined for cell viability and S-nitrosylation of protein disulphide isomerase using the biotin-switch method, and the changes in cell morphology were examined with HE staining. Results METH exposure obviously decreased the cell viability and increased S-nitrosylation of protein disulphide isomerase, and the effect of METH was obviously inhibited by L-NNA treatment. Conclusion METH can cause obvious neurotoxicity and promote S-nitrosylation of protein disulphide isomerase in PC12 cells.