体外培养大鼠星形胶质细胞液压冲击损伤的研究  

The study in primary cultured astrocytes following fluid percussion injury

作　　者：何滔[1,2] 杨术旺[3] 郭赟[1] 赵占考[1] 陈旭义[1] 张永亮[3] 

机构地区：[1]武警部队医学院附属医院 [2]天津市职业与环境危害生物标志物重点实验室,天津300162 [3]武警部队医学院,天津300162

出　　处：《中国应用生理学杂志》2010年第1期46-50,共5页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金资助项目(30471934)

摘　　要：目的:研究大鼠星形胶质细胞液压冲击损伤后形态学及蛋白质组学表达变化。方法:建立体外培养星形胶质细胞液压冲击损伤模型。原代培养SD大鼠脑皮质星形胶质细胞,随机分为损伤组与对照组,对照组给予(0.2±0.01)MPa液压冲击损伤,损伤后不同时间点观察细胞形态学变化;双向凝胶电泳技术分析液压冲击损伤后蛋白质组学表达变化。结果:星形胶质细胞在液压冲击损伤后发生了显著的形态学改变,损伤后2h星形胶质细胞出现了细胞水肿、细胞皱缩、细胞连接断开和坏死,损伤后24h、48h细胞胞体肥大、突起增粗明显,部分区域细胞反应性增生明显。液压冲击损伤后,星形胶质细胞蛋白质表达谱发生了显著改变,损伤后有13个蛋白点表达发生显著改变,其中5种蛋白得到质谱鉴定,分别是肌动蛋白结合蛋白、破解蛋白、磷酸甘油酸变位酶1、NADH脱氢酶10亚基和膜联蛋白1。结论:液压冲击损伤能够引起星形胶质细胞发生显著的形态学改变和蛋白质谱表达改变,损伤后表达改变的蛋白质可能与星形胶质细胞的损伤后应激反应相关。Objective：To investigate the morphology alterations and proteomics changes in primary astrocytes following fluid percussion injury.Methods：Primary cultures of astrocytes were prepared from cerebral hemispheres of 1～3 d-old SD rats,then,astrocytes were randomly divided into control group and injury group which were subjected to （0.2±0.01） MPa fluid percussion injury.The changes of protein expression pattern in astrocytes between injury and control groups were monitored with two dimensional gel electrophoresis.Results：Astrocytes＇s abnormalities of morphology after injury were apparent.The fluid percussion injury caused astrocytes edema,shrinkage,cell junction disconnection and necrosis at 2 h after injury.24 h and 48 h after injury,most part of astrocytes＇s dendrites and soma became hypertrophy and showed a higher rate of cell proliferation.The dynamic proteomics changes were identified and total different 13 spots were detected in this study from the 2DE gels.The different displayed 5 spots were identified via MALDI-TOF：cofilin 1,destrin,phosphoglycerate mutase 1,NADH dehydrogenase （ubiquinone） 1 alpha subcomplex 10,annexin 1.Conclusion：The obvious alteration of morphology and protein expression pattern in primary cultured astrocytes could be induced after fluid percussion injury.The differential proteins detected were probably related to stress responses.

关 键 词：液压冲击损伤 星形胶质细胞 SD大鼠 双向电泳 

分 类 号：R363.22[医药卫生—病理学]