酸性成纤维细胞生长因子对庆大霉素诱导的海马星形胶质细胞MDA、NO、SOD、GSH-Px的影响

摘要

Objective: To study the protective effects of acidic fibroblast growth factor (aFGF) on rats' as-trocytes of hippocampus injured by Gentamicin (GM) in vitro. Methods: After hippocampus were separated, sheared and digested with trypsin, astrocytes were cultured. The third generations were cultured for 3 days in a 24-well culture plate and divided into 3 groups for the experiments (I) control group: normal culture; ?GM group: 2 g/L GM was added for 24 hoursi ?GM plus aFGF intervention group (aFGF + GM) . After adding aFGF, 24 hours, then added GM, 24 hours. Each group contained 6-hole complex and the experiment repeated 3 times. Cell morphology and the changes of MDA, NO, SOD and GSH-Px were observed. Results; Control group; cell morphology was various, three-pole or multi-pole, exhibited strong refraction, processes were intertwined and formed a network; GM group: large number of cells exhibited shrinkage, fusing into a mass, the network and the process connection was destroyed, and much particles and vacuoles existed in cytoplasm) aFGF+GM group: the extent of cell shrinkage and cell fusion were relieved, the damage of process connection was not obvious, and few particles and vacuoles existed in cytoplasm. Comparing GM group with control group, SOD and GSH-Px enzyme activities decreased, while MDA and NO levels increased ( P <0. 01) in GM group, indicating that the cultures of astrocytes and the injured models of GM were successful. The differences in biochemistry and enzymology of the culture con-rntents between GM and aFGF + GM groups were significant (P < 0. 01 ). Conclusion: aFGF could obviously protect damaged astrocytes by GM.%目的:研究酸性成纤维细胞生长因子(aFGF)对庆大霉素(GM)引起的体外培养的大鼠海马星形胶质细胞损害的保护作用.方法:大鼠海马经分离、剪切、胰蛋白酶消化,进行星形胶质细胞的体外培养.传3代的细胞接种于24孔板培养3d后用于实验.实验分3组:①对照组:正常培养；②GM组:2 g/L GM作用24 h；③aFGF+GM组:加入4.25 μg/L aFGF 24 h后再加2 g/L GM培养24 h.每组设6个复孔,实验重复3次.观察细胞形态及丙二醛(MDA)、一氧化氮(NO)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)变化.结果:对照组:细胞形态多样,呈三极、多极型,折光性强,多数细胞突起相互交织成网状；GM组:细胞大量皱缩,聚集成团,网状结构被破坏,突起连接中断,细胞质内出现较多的颗粒和空泡；aFGF+ GM组:细胞皱缩程度减轻,聚集成团现象减少,突起连接中断不明显,细胞质内少见颗粒和空泡.GM组与对照组比较,SOD、GSH-Px酶活性下降,而MDA、NO升高(P＜0.01),提示星形胶质细胞的培养和庆大霉素损伤的建模成功；aFGF+ GM组与GM组比较,各生化和酶学指标差异均有统计学意义(P＜0.01).结论:aFGF对庆大霉素诱导的星形胶质细胞损伤有明显的保护作用.