氯化钴诱导化学缺氧对N9小胶质细胞的损伤作用及机制  

Injury effect of CoCl2-induced chemical hypoxia on N9 microglia

作　　者：王国红[1] 毕凌云[2] 李超堃[1] 侯软玲[1] 尹雅玲[1] 李东亮[1,3] 

机构地区：[1]新乡医学院生理学与神经生物学教研室,河南新乡453003 [2]新乡医学院第一附属医院儿科,河南卫辉453100 [3]复旦大学医学神经生物学国家重点实验室,上海200032

出　　处：《中国药理学通报》2013年第4期557-562,共6页Chinese Pharmacological Bulletin

基　　金：河南省2009年科技发展计划项目(No 092102310098);复旦大学神经生物学国家重点实验室开放课题资助(No 10-10);新乡医学院重点领域开放课题资助(No ZD 2011-2)

摘　　要：目的研究氯化钴(CoCl2)对N9小胶质细胞的损伤作用及机制。方法不同剂量CoCl2处理N9小胶质细胞后,XTT法检测细胞活力;倒置相差显微镜下观察细胞形态变化;硫代巴比妥酸(TBA)显色法检测细胞上清液中丙二醛(MDA)含量;Hoechst 33258染色检测细胞凋亡;流式细胞术检测细胞周期变化。结果 CoCl2(50～1 000μmol.L-1)处理24 h后,细胞活力明显降低(P<0.01),且呈剂量依赖性。细胞形态发生明显改变,部分细胞出现皱缩、破碎,细胞密度减小,随CoCl2剂量增大,细胞形态损伤程度进一步恶化。MDA含量检测结果显示,氯化钴处理后,细胞上清液中MDA含量明显增多(P<0.05)。Hoechst 33258染色结果显示,氯化钴处理组的细胞凋亡比例较对照组明显增多(P<0.01),且比例随CoCl2剂量增大而增多。细胞周期检测结果显示,CoCl2可使细胞周期阻滞于G1/G0期。结论 CoCl2可呈剂量依赖性损伤N9小胶质细胞,机制与诱导细胞凋亡、引发氧化应激反应及细胞周期阻滞有关。Aim To investigate the injury effect of CoCl2 on N9 microglia and explore its mechanism.Methods N9 microglia was exposed to different doses of CoCl2 for 24h,the cell viability was assessed by XTT assay.Cellular morphological changes were observed under phase-contrast microscope.The MDA level in the cultured supernatant was detected with thiobarbituric acid（TBA） method.The cell apoptosis was evaluated by Hoechst 33258 staining.The cell cy-cle was detected by flow cytometry（FCM）.Results CoCl2（50～1 000 μmol·L-1） reduced the cell viability significantly（P＆lt;0.01） in a dose-dependent manner.The morphology of the cells changed obviously after CoCl2 treatment for 24h.Cell shrinkage and necrosis were observed and the cell density decreased.The morphological damage aggravated with the increase of the dose of CoCl2.TBA assay showed that the MDA content in the medium markedly increased（P＆lt;0.05） after CoCl2 treatment.Hoechst 33258 staining showed that CoCl2 increased the percentage of apoptotic cells dose-dependently（P＆lt;0.01）.FCM showed that CoCl2 blocked N9 microglia at G1/G0 phase.Conclusion CoCl2 can damage N9 microglia dose-dependently by inducing cell apoptosis,causing oxidative stress and cell cycle arrest.

关 键 词：氯化钴 诱导 化学缺氧 N9小胶质细胞 细胞损伤 模型 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]