ABSTRACT
BACKGROUND:Hyperbaric oxygen(HBO)treatment promotes the proliferation and differentiation of endogenous neural stem cells in neonatal rats following hypoxic/ischemic brain damage(HIBD).The Wnt signaling pathway is associated with neurogenesis.However,there are few data recording the role of HBO in the differentiation of neural stem cells in vitro.OBJECTIVE:To observe the effect of HBO on differentiation and Wnt3 expression of bone marrow mesenchymal stem cells(BMSCs).METHODS:BMSCs were isoiated and cultured.The rat BMSCs of passages 3-5 were cultured in DMEM/F12(1:1)medium with basic fibroblast growth factor,epidermal growth factor and B27 for 24 hours.The induced BMSCs were randomly divided into two groups:control group(no treatment)and HBO group(HBO,0.10 MPa,60 minutes stabilizing pressure with at least 90% oxygen).The neuron specific encloase(NSE),glial fibrillary acidic protein(GFAP)and 04 marked oligodendrocyte immunocytochemistry were detected by immunofluorescent staining,and Wnt3 protein expression was detected by Western-blot.RESULTS AND CONCLUSION:BMSCs cultured in classic medium of neural stem cells could significantly induce the expression of nestin.The expression of NSE and 04 of HBO group was greater than control group(P＜0.01),but GFAP expression displayed no significant difference between the groups(P＞0.05).Western blot showed HBO could enhance the Writ3 expression (P＜0.05).Results show that HBO can induce BMSCs to differentiate into neural cells and oligodendrocyte,which is correlated with the activation of the Wnt3 protein.