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目的研究叶酸(folate,Fol)对体外培养缺氧神经干细胞(neural stem cell,NSCs)增殖、凋亡和抗氧化能力的影响。方法采用无血清体外细胞培养方法,培养新生大鼠NSCs。将其分为4组,即正常对照(normal control,NC)、缺氧模型(hypoxia,Hyp)、缺氧叶酸缺乏(Hpy+Fol-D)和缺氧叶酸添加(Hpy+Fol)组。除NC组以外,其余三组放入缺氧环境中6h,造成缺氧损伤。采用MTT法检测缺氧后细胞增殖能力的变化;收集增殖6d的细胞,测定缺氧后NSCs超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)的活性;采用Western blot方法检测caspase-3的活性表达水平。结果与NC组比较,缺氧干预后NSCs增殖明显下降,SOD和GSH-PX活性降低,caspase-3的活性表达增加;Hpy组细胞增殖能力、SOD和GSH-PX活性均显著高于Hpy+Fol-D组,低于Hpy+Fol组(P<0.05),caspase-3的活性表达显著高于Hpy+Fol组,低于Hpy+Fol-D组(P<0.05)。结论缺氧可造成NSCs损伤,叶酸缺乏能使缺氧后NSCs的损伤加重,补充叶酸能促进缺氧后NSCs增殖,提高其抗氧化能力,抑制NSCs凋亡,对体外大鼠NSCs的缺氧损伤恢复有促进作用。
Objective To study the effects of folate (Fol) on the proliferation, apoptosis and antioxidant capacity of hypoxic neural stem cells (NSCs) cultured in vitro. Methods The serum-free in vitro cell culture method was used to culture neonatal rat NSCs. The rats were divided into 4 groups: normal control (NC), hypoxia (Hyp), Hpy + Fol-D and Hpy + Fol groups. Except NC group, the other three groups were placed in hypoxic environment for 6h, resulting in hypoxia injury. The proliferation of cells was detected by MTT assay. After proliferation for 6 days, the superoxide dismutase (SOD) and glutathione peroxidase (GSH- Px). Western blot was used to detect the activity of caspase-3. Results Compared with NC group, the proliferation of NSCs was significantly decreased and the activities of SOD and GSH-PX were decreased and the expression of caspase-3 was increased after hypoxia. The cell proliferation, SOD and GSH-PX activities of Hpy group were significantly higher than those of Hpy + Fol -D group, which was lower than that of Hpy + Fol group (P <0.05). The activity of caspase-3 was significantly higher than that of Hpy + Fol group and Hpy + Fol-D group (P <0.05). CONCLUSION: Hypoxia can cause injury of NSCs. Folic acid deficiency can aggravate the damage of NSCs after hypoxia. Supplementation of folic acid can promote the proliferation of NSCs after hypoxia, enhance their antioxidant capacity, inhibit the apoptosis of NSCs and inhibit the hypoxic injury of NSCs in vitro Recovery has a catalytic effect.