胱硫醚-β-合酶/硫化氢系统在鱼藤酮诱导PC12细胞损伤中的变化  

Changes of endogenous cystathionine-β-synthase / hydrogen sulfide system in PC12 cells injury induced by rotenone

作　　者：许岩[1] 马娜[2] 邓树泳[2] 王金全[2] 冯鉴强[3] 孟金兰[2] 

机构地区：[1]南方医科大学中西医结合医院外三科,广东广州510315 [2]广东药学院基础学院生理学系,广东广州510006 [3]中山大学中山医学院生理学教研室,广东广州510080

出　　处：《中国药理学通报》2014年第10期1372-1376,共5页Chinese Pharmacological Bulletin

基　　金：广东省高校优秀青年创新人才培养计划资助项目(No2012LYM_0083);广东省医学科研基金资助项目(NoA2011295);广东省高等学校大学生创新创业训练计划项目(No 1057313016);2013年广东药学院大学生创新性实验计划资助项目

摘　　要：目的动态研究胱硫醚-β-合酶/硫化氢(CBS/H2S)系统在鱼藤酮诱导PC12细胞损伤中的变化。方法鱼藤酮诱导具有多巴胺能神经元特性的PC12细胞损伤作为帕金森病(Parkinson's disease,PD)细胞模型,免疫印迹法(Western blot)检测CBS的表达;亚甲基蓝分光光度计法检测细胞内CBS酶活性及硫化氢(hydrogen sulfide,H2S)的生成;应用CCK-8比色法检测细胞存活率;GSH检测试剂盒检测细胞内GSH含量。结果鱼藤酮损伤6和12 h组,PC12细胞内CBS的表达和活性升高,H2S生成量增加;而在24和48 h组,CBS表达和活性则明显降低,H2S的生成量明显减少;鱼藤酮可以时间依赖性的降低细胞存活率及减少细胞内GSH的含量。结论鱼藤酮刺激引起内源性CBS的表达和活性先升高后降低,H2S的生成先增加后减少,这可能与PC12细胞对抗鱼藤酮诱导的氧化应激损伤有关。Aim To research dynamically the changes of endogenous cystathionine- β-synthase/hydrogen sul-fide system in PC12 cells injury induced by rotenone. Methods Rotenone-induced injury in PC12 cells （ characteristic of dopaminergic neurons） was used as a PD cell model. The expression of CBS was evaluated by Western blot. Intracellular CBS activity and H2 S production were detected by Methylene blue spectro-phot-ometric method. The viability of PC12 cells was measured by CCK-8 assay. GSH detection kit was used to detect the intracellular GSH content. Results In the groups of 6 and 12 hours, the expression and activ-ity of CBS were elevated, and the production of H2 S was increased. In the groups of 24 and 48 hours, CBSexpression and activity were significantly decreased, and the amount of H2 S was significantly reduced. Ap-plication of 1. 5 μmol·L-1 rotenone for different time （6-48h） could decrease the cell viability and intra-cellular GSH contents in a time-dependent manner. Conclusions The expression and activity of endoge-nous CBS, stimulated by rotenone, are elevated firstly and then decreased. The generation of H2 S, stimulated by rotenone, is increased and then reduced significant-ly, which may be related to PC12 cells against oxida-tive stress damage induced by rotenone.

关 键 词：鱼藤酮 帕金森病 PC12细胞 硫化氢 胱硫醚-β-合酶 氧化应激 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R745.7[医药卫生—基础医学]