Objective To explore the effects of transplantation of neural stem cells(NSCs) on connexin 43 (Cx43 expression of traumatic brain injury(TBI) in rats.Methods Thirty-six male Sprague Dawley rats were divided into control group and NSCs transplantation group randomly with 18 cases in each group.Primary NSCs were isolated from the embryonic 14-day (E14) rats and cultured.To induce differentiation of NSCs,a single cell suspension of NSCs was switched into the basic neurosphere medium with 10％ fetal bovine serum(FBS).Immunochemistry staining was conducted to identify anti-uestin for NSCs and anti-β Ⅲ-tubulin for neurons.The model of TBI was build as discribed by modified Feeney.The model rats of TBI received delivery of the NSCs suspension with the stoelting quintessential injector.The expressions of Cx43 in the injuried brain tissue were detected by immunohistochemistry,immunoblotting and reverse transcription polymerase chain reaction(RT-PCR) during different time points.Results Primary NSCs were isolated and cultured as suspension,neurospheres were formed.All the clones were nestin(+).Most cells had ecptoma and extended gradually wit the increasing of arborization at the 2nd day of induction and differentiation of NSCs in vitro,and part of the cells were positive of βⅢ-tubulin for neurons at the 5st day.The immunohistochemistry study showed that brown-yellow Cx43 staining was found in the cell membrane,in the cytoplasm juxtaposed to the cell membrane and in the processes in injured brain with or without NSCs transplantation at 1,2 and 4 weeks.And the Cx43 staining was significantly greater in the NSCs-transplanted rats compared to the control rats at different time points.Significantly higher expressions of gene and protein of Cx43 were found in NSCs-transplanted rats compared to the control rats in the period of 4 weeks post-transplantation.Conclusion Significantly higher expression of Cx43 is found in the transplant core and border of the NSCs-transplanted rats after TBI.%目的 探讨神经干细胞(NSCs)移植对创伤性脑损伤(TBI)大鼠脑组织中连接蛋白43(Cx43)表达的影响.方法 将6～7周龄雄性SD大鼠36只随机分为对照组和NSCs移植组,每组18只.从15只E14大鼠胚胎分离NSCs,进行原代培养及传代培养；用含体积分数10％胎牛血清的培养基对NSCs进行诱导分化；采用免疫细胞化学技术对培养的NSCs及其分化为神经元的表型进行鉴定；采用改良的Feeney法制备TBI模型；利用脑立体定位仪和微量注射泵进行NSCs脑内移植；采用免疫组织化学技术、免疫印迹技术和反转录-聚合酶链反应(RT-PCR)技术检测在移植后不同时间脑组织损伤区Cx43的表达.结果 在培养基中,NSCs呈球团状悬浮生长,神经上皮干细胞巢蛋白表达阳性.NSCs体外诱导分化第2天,多数细胞伸出突起,以后突起逐渐延长,分支增加；分化后第5天,部分细胞βⅢ-微管蛋白阳性.免疫组织化学结果显示,无论有无NSCs移植,在移植后第1、2和4周受损脑组织中,均可见Cx43在细胞膜上、膜旁的细胞质及突起中呈棕黄色表达.NSCs移植大鼠的Cx43染色在各个时间点均显著强于对照组(P＜0.05).在移植后的4周内,NSCs移植组移植点及其周围脑组织中Cx43 mRNA和蛋白表达显著高于对照组(P＜0.01).结论 移植NSCs至TBI大鼠损伤脑组织,在移植点周围脑组织中Cx43的表达显著增加.
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