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[目的]探讨线粒体途径在羧甲基壳聚糖(carboxymethylated chitosan,CMCS)保护过氧化氢(H_2O_2)诱导雪旺细胞(Schwann cells,SCs)凋亡中的作用及机制。[方法]体外培养SCs,S-100免疫荧光染色鉴定。将SCs分为空白对照组、H_2O_2诱导组、H_2O_2加CMCS处理组。流式细胞仪检测细胞凋亡比率,罗丹明(Rhodamine123)荧光染色检测细胞线粒体膜电位水平,Western blot检测SCs内细胞色素C(Cytochrome C)表达水平。[结果]本实验培养细胞经S-100荧光染色鉴定阳性率达95%以上,H_2O_2诱导SCs凋亡,降低线粒体膜电位,增加细胞色素C释放;而在加入CMCS后SCs凋亡比例降低,线粒体膜电位增加,细胞色素C释放减少。[结论]CMCS通过抑制线粒体细胞凋亡途径保护H_2O_2诱导SCs凋亡。
[Objective] To explore the role and mechanism of mitochondrial pathway in the apoptosis of Schwann cells (SCs) induced by carboxymethylchitosan (CMCS) and hydrogen peroxide (H 2 O 2). [Method] SCs were cultured in vitro and identified by S-100 immunofluorescence staining. SCs were divided into blank control group, H 2 O 2 induction group, H 2 O 2 plus CMCS treatment group. Flow cytometry was used to detect the ratio of apoptosis, Rhodamine123 fluorescence staining was used to detect the level of mitochondrial membrane potential, and Western blot was used to detect the expression of Cytochrome C in SCs. [Results] The positive rate of the cultured cells was above 95% by S-100 fluorescent staining, H 2 O 2 induced the apoptosis of SCs, decreased the mitochondrial membrane potential and increased the release of cytochrome C. The percentage of apoptosis of SCs decreased after addition of CMCS. Increased membrane potential, cytochrome C release decreased. [Conclusion] CMCS can protect HUVEC from inducing apoptosis of SCs by inhibiting mitochondrial apoptosis pathway.