碱性成纤维生长因子对体外培养鼠胚胎中脑神经细胞缺氧反应的影响

Effect of basic fibroblast growth factor on anoxia of rat embryo midbrain nerve cells cultured in vitro

目的：观察不同浓度碱性成纤维生长因子（bFGF）对胚胎中脑神经细胞缺氧的保护作用。方法：14 d的大鼠胚胎中脑取材,分离、消化后作原代培养,分为对照组,单独缺氧组,不同浓度的（1.0,5.0,10.0,50.0,100.0 ng/mL）bFGF＋缺氧组;采用羟胺比色法对超氧化物歧化酶（SOD）活性进行测定,采用硫代巴比妥酸比色法进行丙二醛（MDA）测定;采用考马斯亮蓝法进行相对蛋白质含量测定,同时进行细胞形态学观察。结果：单独缺氧组中脑神经细胞MDA含量升高,SOD活性、相对蛋白质含量及细胞集落存活率下降,与对照组比较（P〈0.01）,并出现细胞形态学损伤;5.0~50.0 ng/mL bFGF＋缺氧组中脑神经细胞MDA含量下降,SOD活性、相对蛋白质含量、细胞集落存活率升高,与单独缺氧组比较（P〈0.01）,细胞无明显形态学损伤;当bFGF剂量为1.0 ng/mL和100 ng/mL时神经细胞MDA含量、SOD活性、相对蛋白质含量及细胞集落存活率与单独缺氧组比较无明显改变（P〉0.05）,细胞有明显的形态学损伤。结论：适当的bFGF剂量能对胚胎中脑神经细胞缺氧损伤具有明显保护作用。

Objective：To investigate the protection effect of different densities of basic fibroblast growth factor（bFGF） on anoxia of the embryo midbrain neurocytes cultured in vitro.Methods：Primary culture of embryo midbrain neurocytes from rats at 14 old was performed after being separated and digested,and then the cells were divided into control group,group in anoxia,different densities bFGF treated group.Superoxide dismutase（SOD） activity was determined by hydroxylamine colorimetric method,malondialdehyde（MDA） by thiobarbituric acid colorimetric method and relative protein content by coomasiee brilliant blue.The morphological observation was simultaneously performed.Results：In the neurocytes of the anoxia alone group,MDA content was increased,SOD activity,relative protein content and colony survival rate were decreased compared with control group（P0.01）,morphological damage occurs;in the anoxia group treated with 5.0-50.0 ng/mL bFGF,MDA content was decreased,SOD activity and relative protein content and colony survival rate were increased compared with anoxia alone group（P0.01）,showing no obvious morphological cell damage;when the bFGF dose of 1.0 ng/mL and 100 ng/mL,the MDA content,SOD activity,relative protein content and colony survival had no significant change compared with anoxia alone group（P0.05）,showing obvious morphological damage in cells.Conclusion：The appropriate dose of bFGF can have a significant protective effect of hypoxic injury in embryonic midbrain neurocytes.