星形胶质细胞对损伤反应的体外实验研究

Experimental study of astrocyte reaction to injury in vitro

目的 研究体外星形胶质细胞 (Ast)反应性胶质化的发生、发展过程与规律。方法 体外分离培养Ast,利用划伤的方法 ,建立Ast对损伤反应的实验模型 ,通过形态学观察、RT PCR、免疫细胞化学、原位杂交及图像分析等方法 ,研究体外Ast对损伤的反应及规律。结果 损伤体外培养的Ast后 ,观察到反应性胶质化的典型特征 ,表现为Ast胞体肥大、突起增粗、延长 ,GFAP免疫细胞化学染色增强 ;原位杂交、RT PCR分析表明 ,GFAPmRNA表达显著增高。上述变化在伤后 1d即可见到 ,5～ 7d达到高峰。结论 体外分离培养的Ast对损伤表现出活跃的反应 ,呈现出典型的反应性胶质化特征 ,其发生。

Objective To study the occurence, development and regulation of reactive gliosis with astrocyte (Ast) in vitro . Methods Ast was isolated and cultured in vitro and its model of reactive gliosis was established by scratching the cultured astrocytes. The reactivity and rules of Ast to injury was studied by morphological changes, RT PCR, immunocytochemistry, in situ hybridization and imaging analysis. Results After scratching, the astrocytes showed typical features of reactive gliosis, with the hypertrophic cell body, thickened and lengtheded processes, and enhanced glial fibrillary acidic protein (GFAP) staining. In situ hybridization and RT PCR analysis confirmed that the expression of GFAP mRNA was markedly increased. These changes occurred 1 d after scratching and reached the peak 5 to 7 d after injuring. Conclusion A model of reactive astrogliosis was successfully established in vitro which showed an active reaction to injury. The characteristics of reactive gliosis parallel that seen in vivo .