ABSTRACT
BACKGROUND: Previous studies have verified that under normal culture of neural stem cells (NSCs), folic acid can accelerate proliferation of NSCs by phosphorylation of mitogen activated protein kinase path activation ERK1/2. OBJECTIVE: To investigate the effect of folic acid on NSC extracellular signal regulatory protein kinase pERK1/2 under hypoxic condition. METHODS: NSCs from Neonatal rats were cultured in vitro by serum-free culture method, and incubated in a flask at 1×10~8/L. Except normal control group, self-made hypoxia equipment was used in the hypoxia model, folic acid deficiency and folic acid supplemented groups at day 3. At 37 ℃, hypoxia culture was conducted in the thermostat for 6 hours. The contents of folic acid were 4 mg/L, 4 mg/L, 0.65 mg/L, 8 mg/L in the four groups. Cells following 6 days were collected to count the density using trypan blue. RT-PCR was utilized to detect pERK1/2 mRNA expression. Western blot assay was employed to determine pERK1/2 protein expression. RESULTS AND CONCLUSION: Compared with the normal control group, the proliferation of NSCs and the expression of ERK1/2mRNA and pERK1/2 protein were decreased significantly in the hypoxia model group. Compared with the hypoxia model group, the proliferation of NSCs and the expression of ERK1/2mRNA and pERK1/2 protein were increased in the folic acid supplemented group, whereas decreased in the folic acid deficiency group. There were significant differences among groups (P < 0.001). Above-described results verified that folic acid supplementation can activate ERK1/2 phosphorylatin and accelerate proliferation of NSCs under hypoxia condition.
ABSTRACT
Objective To study the effects of folate(Fol) on proliferation,apoptosis and the antioxidative capacity of the NSCs under hypoxia in vitro.Method The NSCs taken from infant rats brain were cultured by serum-free medium in vitro and divided into four group which were normal control group(NC),hypoxia group(Hyp),Hpy+Fol-D(folate deficient)group and Hpy+Fol group.Except NC group,the other groups were cultured in hypoxia condition for 6 h.The proliferation of NSCs after hypoxia damage was detected by MTT test.The NSCs were collected after being cultured 6 d and then the activity of SOD and GSH-PX was examined.The expression of activated caspase-3 was detected by Western blot.Results Compared with NC group,the proliferation of NSCs and the activity of SOD and GSH-PX decreased significantly after hypoxia,while the expression of activated caspase-3 increased0 The proliferation of NSCs and the activity of SOD and GSH-PX in Hyp group were significantly higher than Hyp+Fol-D group but much lower than Hpy+Fol group(P