高压氧对缺氧缺血性脑损伤新生SD大鼠神经干细胞增殖分化的影响

Influence of hyperbaric oxygen therapy on rats neural stem cells proliferation after hypoxic-ischemic brain damage

目的探讨在离体细胞水平下高压氧(HBO)对缺氧缺血性脑损伤(HIBD)时新生SD大鼠神经干细胞(NSCs)增殖分化的影响。方法采用Rice-Vannucci法制成HIBD模型,分别制备正常及HIBD时SD大鼠脑组织匀浆,将传至2～3代的SD大鼠NSCs根据处理的不同随机分为空白对照组(CON组)、与正常脑组织匀浆共培养组(NC+N组)、与HIBD脑组织匀浆共培养组(NC+HIBD组)、与HIBD脑组织匀浆共培养1h后给予HBO处理组(NC+HIBD+HBO组)和与正常脑组织匀浆共培养1h后给予HBO处理组(NC+N+HBO)5组。5组细胞在同一时间行免疫荧光染色,分别进行NSE、GFAP、O4染色,荧光显微镜下计数各组NSCs分化为NSE阳性细胞、GFAP阳性细胞和O4阳性细胞的百分率,并进行比较。结果与CON组相比,NC+N+HBO组、NC+N组、NC+HIBD组分化为神经元和少突胶质细胞的百分比明显增加(P<0.05),其中HIBD组增加最多,在此基础上再给予HBO处理,NSCs向神经元的分化会进一步增加,向少突胶质细胞的分化也相应增加;与CON组相比,NC+N组、NC+HIBD组、NC+HIBD+HBO组分化为星形胶质细胞的百分比明显减少(P<0.05)。结论在离体细胞水平,HBO可促进HIBD大鼠NSCs分化为神经元和少突胶质细胞而抑制其向星形胶质细胞分化。

Objective To investigate the influences of hyperbaric oxygen（HBO） and rat brain extracts on proliferation of rats neural stem cells in vitro.Methods Rats NSCs were randomly divided into five groups respectively according to the treatments as follows： blank control group（CON group）： without any treatment;NC＋N group： incubated with normal brain tissue extract conditioned cultures;NC＋HIBD group： incubated with HIBD brain tissue extract conditioned cultures;NC＋HIBD＋HBO group： incubated with HIBD brain tissue extract conditioned cultures 1 h then treated with HBO;NC＋N＋HBO group： incubated with normal brain tissue extract conditioned cultures 1 h,then treated with HBO.Immunocytochemical staining was performed simultaneously on the 5 groups of NSCs.NSE-positive NSCs,O4-positive NSCs and GFAP-positive NSCs were counted for percentage of the positive cells′.Results Compared with CON,NC＋N＋HBO group,NC＋N group the NC＋ of HIBD percentage of cells which differentiated into neurons and oligodendrocytes increased significantly（P0.05）,which HIBD group increased the most.On this basis,HBO treatment was given,differentiation of NSCs to neuronal would be further increased,there was a corresponding increase in the differentiation of oligodendrocytes;compared with CON,NC＋N group,NC＋HIBD group,NC＋ of HIBD＋HBO group differentiation,the percentage of astrocytes significantly reduced（P0.05）.Conclusion HBO could promote ncNSCs differentiate into neuronal or oligodendrocyte,and inhibit ncNSCs differentiate into astrocytes.