体外培养大鼠大脑皮质星形胶质细胞损伤后bFGF的表达

In Vitro Study on bFGF Expression of Rat Astrocytes after Mechanical Injury

目的观察体外培养星形胶质细胞损伤后碱性成纤维细胞生长因子(basic fibroblast growth fac-tor,bFGF)的表达变化,进一步了解脑损伤修复的分子机制,以期为法医病理学鉴定提供更多的依据。方法取出生后24h内SD大鼠大脑皮质进行星形胶质细胞体外分离培养和纯化,将其随机分为对照组,损伤后30min和1、3、6、12、24h及3、7d组,应用ABC法检测不同时间段bFGF表达的差异。结果体外培养星形胶质细胞的纯度达95%以上。对照组有少量的bFGF蛋白表达。机械性损伤后1～3h,bFGF反应强度开始有变化,6～12h明显增强,24h达高峰,3d以后开始下降。结论机械性划痕损伤后bFGF表达与在体动物实验模型一样具有时序性变化规律,仅表达时间提前,说明损伤后bFGF表达可为脑损伤时间的推断依据之一;体外培养细胞损伤模型对组织细胞损伤的分子机制研究具有一定意义。

Objective To study the alteration of basic fibroblast growth factor（bFCF） expression in astrocytes in vitro after mechanical injury and to understand the repair mechanism of brain injury. Methods Astrocytes were isolated from cerebral cortex of SD rats born in 24 hours, and then cultured and purified. The cultured astrocytes were randomly divided into control group and injury groups that were subjected to mechanical injury at 30 min, 1 h, 3 h, 6 h, 12 h, 24 h, 3 d and 7 d. The levels of bFGF expression in the astroeytes after injury were detected by ABC immunohistochemistry. Results More than 95% of the cultured cells were as- trocytes. The levels of bFGF expression were very low in the control group. On the other hand, increased levels of bFGF expression could be observed at 1-3 h after injury. The expression levels increased significantly at 6-12h, reached peak level at 24h, remained at the high level up to 3 days, and then deereased gradually. Conclusion The changes of bFGF expression levels in cultured astrocytes in vitro after mechanical injury are similar to that observed in vivo experimental model, both of which show time-dependent characteristic, with only slightly earlier expression of bFGF observed in vitro. Thus, the expression of bFCF after injury can be one of evidences for estimation of brain injury intervals. The cell injury model in vitro may have superiority in the study of the molecule mechanism of tissue and cell injury.