蝙蝠葛酚性碱抗神经元缺血再灌注损伤的作用机制  

Protective Effect of Phenolic Alkaloids from Menispermum dauricum on Neuronal Ischemia/Reperfusion Injury

作　　者：段秋红[1] 王忠强[2] 王剑飞[1] 卢涛[1] 国艳春[1] 郭莲军[3] 王西明[1] 

机构地区：[1]华中科技大学同济医学院基础医学院生物化学与分子生物学系,武汉430030 [2]广州军区武汉总医院急诊科,武汉430070 [3]华中科技大学同济医学院基础医学院药理学系,武汉430030

出　　处：《华中科技大学学报（医学版）》2008年第2期141-144,148,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30570555)

摘　　要：目的探讨蝙蝠葛酚性碱(PAMd)抗神经元细胞缺血再灌注损伤的作用机制。方法体外培养小鼠神经母细胞瘤细胞株N2a,缺糖、缺氧和缺血清(OGSD)培养90 min,更换正常培养液并加入不同浓度的PAMd,继续培养不同时间后,收集细胞培养液,测定乳酸脱氢酶(LDH)活性和神经递质含量;收集细胞,测定以下指标:①MTT法检测细胞生存活力;②荧光光度法分析胞质活性氧(ROS)含量;③激光共聚焦分析线粒体跨膜电位;④Western blot检测胞质细胞色素C(Cyt C);⑤采用Caspase 3可见光底物DEVD-pNa测定Caspase 3活性。结果①PAMd促进OGSD处理后的N2a细胞存活;②PAMd抑制OGSD处理后的N2a细胞释放兴奋性神经递质;③PAMd直接清除细胞内的ROS;④PAMd能维持线粒体跨膜电位的稳定;⑤PAMd抑制线粒体Cyt C释放和抑制胞质Caspase 3活性。结论PAMd通过阻断线粒体凋亡途径和抗氧化作用发挥神经元保护作用。Objective To explore the protective effect of phenolic alkaloids from Menispermum dauricum （PAMd） on neuronal ischemia/reperfusion injury. Methods After N2a cells cultured in vitro were deprived of glucose, serum and oxygen （OGSD） for 90 min, the different concentrations of PAMd were added to the medium. Then, treated cells were cultured for different intervals. At the end of the treatment, the collected culture solution was used for the analysis of the activity of lactate dehydrogenase （LDH） and content of neurotransmitters and the cells were used for the measurements of the following parameters： cell viability （MTT）, reactive oxygen species （ROS） production （fluorophotometry）, mitochondrial transmembrane potential （confocal laser microscopy）, cytochrome C （Western blot） and caspase 3 activity （DEVD-pNa）. Results PAMd promoted OGSD-insulted N2a cell survival. It could not only inhibit exciting neurotransmitters release and directly scavenge ROS, but also maintain △ψm stability, reduce cytochrome C release and decrease caspase 3 activity. Conclusion PAMd could attenuate the injury induced by OGSD by blocking mitochondrial apoptosis pathway and antioxidation, thus it might play a neuroprotection in cerebral ischemia/reperfusion injury.

关 键 词：蝙蝠葛酚性碱 神经元 缺血再灌注损伤 细胞凋亡 细胞色素C CASPASE 3 

分 类 号：R743[医药卫生—神经病学与精神病学]