caspase-3限制小鼠脑卒中恢复期齿状回神经前体细胞增殖  

Restricted effect of caspase-3on proliferation of neuronal precursor cells in dentate gyrus of mice during stroke recovery

作　　者：朱西民 戴义芹 徐昊辰 孙春刚 范文英[1,2] 赵冰樵[1,2] 

机构地区：[1]复旦大学基础医学院医学神经生物学国家重点实验室 [2]复旦大学脑科学研究院脑科学协同创新中心,上海200032

出　　处：《复旦学报（医学版）》2016年第3期280-285,共6页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金重点项目(81530034);国家自然科学基金面上项目(81071062;81271457)~~

摘　　要：目的探讨caspase-3对脑卒中恢复期海马齿状回(dentate gyrus,DG)神经前体细胞(neuronal precursor cells,NPCs)增殖的作用。方法构建小鼠大脑中动脉远端堵塞(distal middle cerebral artery occlusion,dMCAO)缺血模型,取缺血手术后第7天缺血侧DG进行体外细胞培养。研究分为对照组和给药组(caspase-3抑制剂给药组);采用Western blot、免疫细胞化学、流式细胞仪和TUNEL染色检测DG NPCs中caspase-3的表达及NPCs的凋亡和增殖情况。在小鼠脑卒中后第14天,采用免疫组织化学方法检测caspase-3抑制对DG NPCs增殖的影响。结果体外培养的源自脑缺血后DG的神经球NPCs大量表达激活型caspase-3,但是超过87%的caspase-3阳性细胞与TUNEL阳性没有共定位关系。caspase-3限制了DG NPCs的自我更新,但并未参与细胞的凋亡过程。抑制caspase-3可促进脑卒中恢复期DG NPCs的增殖。结论 caspase-3可通过细胞凋亡之外的途径调控缺血性脑损伤后的神经再生。Objective To investigate the effect of caspase-3 on the proliferation of neuronal precursor cells（NPCs）in the dentate gyruse（DG）during stroke recovery. Methods The stroke model was produced by electrocoagulating the distal middle cerebral artery（dMCA）of mice,then the NPGs were cultered from ischemia DG on the 7th day after stroke.Treatment groups and control groups were assigned in a randomized manner.The expression of caspase-3 and apoptosis and proliferation of NPCs were analyzed by Western blot,immunocytochemistry,flow cytometry analysis and TUNEL staining in NPCs cultured from ischemic DG.Besides,the effects of caspase-3inhibition on the proliferation of NPCs in the DG was also investigated by immunohistochemistry in the mouse at 14 day after stroke.Results Caspase-3 was expressed in cultured NPCs,but more than87% of the caspase-3 positive cells and TUNEL positive cells were not collocated.Caspase-3 negatively regulated self-renewal of NPCs,but did not participate in cell death in culture DG NPCs.Caspase-3 negatively regulated NPCs proliferation in the DG during stroke recovery. Conclusions Caspase-3 may possess a novel function that limits the neurogenic response after stroke.

关 键 词：脑卒中 神经再生 CASPASE-3 齿状回 神经前体细胞 小鼠 

分 类 号：R363[医药卫生—病理学]