叶酸对缺氧新生鼠神经干细胞体外增殖的影响

Effects of folic acid on proliferation of NSCs of neonatal rats after hypoxia

目的:探讨叶酸(FA)对缺氧条件下体外新生大鼠神经干细胞(NSCs)增殖的影响。方法:取24h内新生SD大鼠大脑半球,进行NSCs原代培养。分为正常对照组、缺氧模型组、缺氧叶酸添加组、缺氧叶酸缺乏组共4组。培养第3天除正常对照组外其他3组进行缺氧处理,造成缺氧损伤;培养6d后收集细胞,进行MTT法、5-溴脱氧尿苷(BrdU)掺入法和免疫组化双标记法检测叶酸对缺氧NSCs增殖的影响。结果:MTT检测显示,正常对照组细胞OD值较缺氧模型组高,差异有统计学意义(P<0.05);缺氧叶酸添加组OD值明显高于其它缺氧组,差异均有统计学意义(P<0.05);免疫组化双标阳性率比较,缺氧模型组双标阳性率明显下降,与正常对照组比较差异有统计学意义(P<0.05),缺氧叶酸添加组明显高于其它几组,差异均有统计学意义(P<0.05)。结论:长时间持续缺氧可造成神经干细胞缺氧损伤;添加叶酸使缺氧神经干细胞增长率增加,提示叶酸能促进体外缺氧新生鼠神经干细胞的增殖。

Objective： To investigate the effect of folic acid on neural stem cells （NSCs） proliferation of neonatal rats after hypoxia in vitro. Methods： NSCs were isolated from the brain of neonatal SD rats within 24 hours for primary culture and divided into four groups which were normal control group, hypoxia model group, hypoxia FA-supplement group and hypoxia FA-deficiency group. The NSCs of the hypoxia groups were exposed to hypoxia condition for 6 h at the third day, then collected after culture for 6 d. Proliferation of NSCs were detected by M＇FF, BrdU incorporation assay and double immunocytochemisty method. Results： MrIT showed the OD value in normal control group were higher than that in hypoxia model group （P〈0.05）. The OD value in hypoxia FA-supplement group was significantly higher than that in hypoxia model group and hypoxia FA-deficiency group （P〈0.05）. The percentage of Nestin/BrdU positive cells in total cells in hypoxia model group was decreased than that in normal control group （P〈0.05）, and in hypoxia FA- supplement group the percentage was higher compared with the other two groups （P 〈0.05）.Conclusion： Folic acid could promote proliferation of NSCs of neonatal rats after hypoxia.