Research Of SDF-1 On Effects Of Neural Stem Cells In Vitro Hypoxia And The Mechanism

Objective: By hypoxia incubator to establishe hypoxic model of neural stem cells(NSC),Study of stromal cell-derived factor-1(SDF-1) on neural stem cells influence expression of CXCR4 in vitro hypoxia, explore SDF-1 on NSCs of hypoxic conditions neuroprotective effects.Methods: Cultured C17.2 neural stem cell line as the research object and incubated by hypoxia incubator to mimic the hypoxia injury in vitro. Part 1 : Div-ided into two groups: the normal group, the hypoxia treatment group, MTT was performed to examine whether hypoxia influence the survival rate of NSCs and to explore the optimal time to injure NSCs at 0h,24 h, 48 h,72h time points.AO/EB double fluorescence staining was performed to observe the cellular morphology of NSCs at 0h,24 h,48h time points. Western Blot to detect the expression of HIF-1?. Part 2 : Then divided into two group: the normal control group,the hypoxia treatment group. MTT cell proliferation assay was performed to examine whrther SDF-1 protect the NSCs from hypoxic.Part 3 : the normal control group,the hypoxia treatment group, the SDF-1 treatment group, the SDF-1 with the CXCR4 antagonist AMD3100 treatment group and the AMD3100 treat-ment group.Western Blot to detect the expression of CXCR4,bcl-2,bax protein in each group.Results: Part 1 : The survival rate and cellular morphology of NSCs:a. The results indicated that the 24,48,72 hours hypoxic groups cell survival rate compare to the normal group were decline(P<0.05). AO/EB fluorescent staining: The results indicated that the hypoxic group cell quantity compare to the normal group were decline(P<0.05), and observe the late stage apoptotic cells.b. HIF- 1 alpha protein expression: Under 24 h hypoxic condition, the expression of HIF- 1 alpha protein compared with the normal group increased obviously(P<0.05) ? Under 48 h hypoxic condition, the expression of the HIF- 1 alpha protein increased obviously(P<0.05).Part 2 : Cell survival of NSCs: Under hypoxic conditions for 24 h and 48 h, the SDF-1 treatment group compared with the hypoxic treatment group promoted cell survival rate(P<0.05).Part 3 : The expression of bcl-2,bax,CXCR4 under the hypoxic condition through Western Blot:CXCR4 protein expression: The expression of CXCR4 protein of SDF-1 treatment group was increased compared with the hypoxia treatment group(P<0.05). The expression of CXCR4 protein of hypoxia treatment group was increased compared with the normal group(P<0.05).bcl-2 protein expression: The expression of bcl-2 protein of SDF-1 treatment group was increased compared with the hypoxia treatment group(P<0.05).The expression of bcl-2 protein of hypoxic treatment group was decreased compared with the normal group(P<0.05).bax protein expression:The expression of bax protein of SDF-1 treatment group was decreased compared with the hypoxia treatment group(P<0.05).The expression of bax protein of hypoxic treatment group was increased compared with the normal group(P<0.05)Conclusions: SDF-1 can inhibited the apoptosi of NSCs induced by hypoxic condition, promote the survival rate of NSCs in hypoxic conditions, the mechanism may be related to SDF-1 can regulate bcl-2 and bax.