羧甲基壳聚糖通过抑制线粒体途径保护氧化损伤诱导雪旺细胞凋亡  

Neuroprotective effects of carboxymethylated chitosan on hydrogen peroxide induced apoptosis by inhibiting the mitochondrial signaling pathway in Schwann cells

作　　者：贺斌[1] 陶海鹰[1] 卫爱林[1] 李孝海[1] 陈任[1] 潘峰[1] 郑啸 

机构地区：[1]武汉大学人民医院骨科,武汉430060 [2]武汉市新洲区人民医院骨科,武汉430400

出　　处：《中国矫形外科杂志》2016年第20期1892-1897,共6页Orthopedic Journal of China

基　　金：国家自然科学基金资助项目(编号:81301056);湖北省自然科学基金资助项目(编号:2013CKB002;2015CKB753)

摘　　要：[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] An in vitro model was designed to investigate the mechanisms of carboxymethylated chitosan （CMCS） against oxidative stress induced apoptosis in Schwann cells （SCs）. [Method] SCs were cultured in vitro and identi- fied by S- 100 immunofluorence staining. SCs were divided into blank control group,H202 induced group,as well as H202 ＋ CMCS treated group. Cell apoptotic rate was detected by flow cytometry, mitochondrial transmembrane potential was detected by Rhodamine （Rho123） immunofluorence staining, cytochrome C （CytC） protein level was detected by Western blot analysis. [ Result] In this study, the S -100 positive cells were more than 95%. In H202 induced group, SCs apoptotic rate increased, mi- tochondrial transmembrane potential decreased, and CytC protein level increased. On the contrary, after treated with CMCS in the H2 02 ＋ CMCS group, apoptotic rate decreased, mitochondrial transmembrane potential increased, and CytC protein level de- creased. [ Conclusion] CMCS protects SCs form oxidative induced apoptosis by inhibiting mitochondrial signaling pathway.

关 键 词：羧甲基壳聚糖 雪旺细胞 凋亡 线粒体 氧化应激 

分 类 号：R745[医药卫生—神经病学与精神病学]