Smad4介导Activin A信号在PC12细胞OGD损伤中作用研究  

Effects of ActivinA signaling mediated by Smad4 in PC12 cell suffered by OGD

作　　者：尤如芹 王姣琦[1] 刘洪雨[1] 崔杨[1] 何金婷[1] 纪秋野[2] 刘永峰 莽靖[1] 徐忠信[1] 

机构地区：[1]吉林大学中日联谊医院神经内科,吉林长春130033 [2]吉林大学中日联谊医院科学研究中心,吉林长春130033 [3]美国奥尔巴尼医学院

出　　处：《中国实验诊断学》2015年第4期525-528,共4页Chinese Journal of Laboratory Diagnosis

基　　金：国家自然基金面上项目(81371298);吉林省科技发展计划国际合作项目(20120723);吉林省产业技术研究与开发项目(2013C030-4);吉林大学白求恩前沿交叉学科创新项目(2013107028);吉林大学白求恩医学青年科研基金项目(2013207052)

摘　　要：目的明确OGD损伤诱导的PC12细胞激活素A(Activin A,ActA)信号在细胞内基于Smad4的转导模式及作用。方法使用鼠神经生长因子(NGF,50ng/ml)诱导大鼠嗜铬细胞瘤PC12细胞向神经元样转化,利用无糖DMEM培养液联合连二亚硫酸钠(1mmol/L)构建神经元氧糖剥夺(oxygen-glucose deprivation,OGD)模型,体外模拟神经元缺血性损伤。OGD 0、3h、6h、12h后Realtime-PCR检测ActA mRNA表达水平;激光共聚焦观察免疫荧光染色后的Smad4质核分布情况。结果 NGF诱导后PC12细胞贴壁并伸出较长的突触,向神经元样转化。细胞OGD处理3h后ActA mRNA表达水平较0h显著升高,6h开始下降;OGD3h后Smad4在细胞核内的分布较OGD 0h明显增加,6小时开始下降。结论 OGD损伤可诱导神经元样PC12细胞ActA信号激活,Smad4可通过质核移位介导ActA信号在受体水平下游的信号转导,且随时间延长呈动态变化。Objective To explore the transduction mechanisms and function of the ActivinA （ActA）signaling based on Smad4 in the oxygen-glucose deprivation （OGD）PC12 cell line.Methods Nerve growth factor （NGF,50ng/ml） was used in this study to induce the differentiation of rattus PC12 pheochromocytoma cells to nerve-like cells.Then the glucose-free DMEM and hyposulfite of Na2 S2 O4 （1 mmol/L）was introduced to cells to construct an OGD model in vitro.The expression of ActA mRNA in OGD 0 h/3 h/6 h/12 h group was abserved by Realtime-PCR.And after im-munofluorescence staining,the intranuclear mobility of Smad4 in OGD 0 h/3 h/6 h/12 h group was observed under the confocal laser scanning microscope at 554 nm.Results PC12 cell was successfully differentiated into nerve-like cell af-ter NGF exposure.Compared to the group of OGD 0 h,the expression of ActA mRNA in OGD 3 h group was signifi-cantly increased,while gradually decreased in OGD 6 h and 12 h group.The fluorescence intensity of Smad4 in nuclei was increased in OGD 3 h group compared to that in OGD 0 h group,and decreased in OGD 6h group.Conclusion OGD injury could activate the ActA signaling in nerve-like PC12 cell line.Smad4 could mediate the tranduction of ActA signaling through cytoplasmic-nuclear translocation.

关 键 词：脑缺血损伤 激活素A SMAD4 

分 类 号：R741[医药卫生—神经病学与精神病学]