促红细胞生成素对星形胶质细胞损伤的保护作用

目的探讨促红细胞生成素（EPO）对体外培养的星形胶质细胞的作用。方法采用出生3 d内的SD大鼠大脑制成细胞悬液后,在含15%胎牛血清的DMEM培养液中培养,通过摇床和逐渐传代纯化星形胶质细胞,分为正常组、正常E组（正常细胞＋EPO 10 U/L）、损伤组（缺氧3 h）、损伤E组（损伤细胞＋EPO 10 U/L）。用免疫荧光技术鉴定星形胶质细胞,MTT法检测细胞活性和凋亡情况,PCR技术测定细胞纤维酸性蛋白（GFAP）。结果星形胶质细胞摇床后传至第3代,经鉴定其星形胶质细胞纯度达95%以上;缺营养3 h后部分细胞形态变圆甚至死亡,漂浮在正常细胞表面;正常E组和损伤E组细胞在分别加入EPO 3 d后,损伤E组细胞活性与GFAP表达均明显升高,正常E组则没有明显变化。结论EPO对缺营养损伤的星形胶质细胞有显著的保护作用,而对正常星形胶质细胞无保护作用。

Protective effect of erythropoietin on astrocytes

Objectives To investigate the protective effect of erythropoietin（EPO） on astrocytes（AST） culturing in vitro.Methods Cell suspension made with 3rd-born SD rat′s brain,was cultured by DMEM culture medium containing 15% fetal bovine serum.AST in above-mentioned cell suspension was purified through shaking and passaging and was divided into normal group,normal E group（normal＋EPO 10 U/L）,injury group,injury E group（injury＋EPO 10 U/L）.The AST was identified by immunofluorescence technique.The cytoactive and apoptosis were detected by MTT and the express of GFAP was determined with PCR technique. Results AST was passaged to the 3rd generation of post-shaker. The purity of the astroeytes was more than 95%. Part of cells became round and even died, floated in the surface of normal cells after nulli- nutrition for 3 h. After applying EPO 3rd to normal group and injury group,the cell activity and the expression of GFAP were significantly increased in injury group,but the normal group did not change significantly. Conclusion Appropriate doses of EPO have significant protective effect to the lack of nutritional injury AST, whereas no significant protective effect to normal AST.