Study on regulating of PFT-a to the PUMA in neural stem cells transplantation after cerebral ischemia

Objective To explore the effect of p53 inhibitor(PFT-a) on neural stem cells after cerebral ischemia in p53 downstream apoptotic gene(PUMA) expression.Methods Using SD rat to isolate NSCs,through the primary and passage culture,by oxygen glucose deprivation(OGD) treatment for 6 h,they were divided into OGD + dimethyl sulfoxide(DMSO) group and OGD + PFT-a group,changes of expression of the p53 protein and PUMA by immunoblotting technique were compared between the 2 groups.In vivo tests using Longa method to establish the CT confirmed the brain artery embolism model in 48 rats,they were randomly divided into PFT-a + NSCs transplantation group(n =24) and DMSO + NSCs transplantation group(n =24);using immunofluorescence to detect the PUMA expression in the different groups of transplantation.Results(1) In vitro Oxygen glucose deprivation after 6 h,OGD + PFT-a group's p53 protein into the nucleus level decreased,expression is up-regulated in the cytoplasm,while OGD + DMSO group was mainly expressed in the cell nucleus;the expression level of PUMA was significantly lower than that in OGD + DMSO group(P 0.05);(2) In vivo experiment,compared with the DMSO + NSCs group,PFT-a + NSCs PUMA transplantation group's positive expression cells was significantly reduced[(38.9 ±4.3)%vs.(29.4 ± 5.6)%,P 0.01].Conclusion The PFT-a combined with neural stem cells can inhibit the expression of PUMA,which may be an important mechanism of PFT-a to promote cell survival in the cerebral ischemic area after the transplantation of neural stem.