人脂肪组织来源的神经干细胞移植对大鼠局灶性脑缺血再灌注后细胞凋亡及Bcl-2、Bax蛋白表达的影响  

Effects of transplantation of neural stem cells originated from human adipose tissue on the cell apoptosis and the expression of protein Bcl-2 and Bax following focal cerebral ischemia-reperfusion in rats

作　　者：刘斌[1] 董静[1] 李建民[2] 刘宁[1] 王瑞敏[3] 张晋霞[1] 李世英[1] 陈贵良[4] 

机构地区：[1]华北煤炭医学院附属医院神经内一科,唐山063000 [2]华北煤炭医学院附属医院神经外科,唐山063000 [3]华北煤炭医学院实验中心 [4]华北煤炭医学院实验动物中心

出　　处：《临床神经病学杂志》2010年第1期42-45,共4页Journal of Clinical Neurology

基　　金：河北省自然科学基金(C2008000994);河北省优秀专家出国培训基金(2007);唐山市科研基金(08130204a-1-21)

摘　　要：目的探讨人脂肪组织来源的神经干细胞移植对大鼠局灶性脑缺血再灌注后细胞凋亡及Bcl-2、Bax蛋白表达的影响。方法线栓法制作大鼠大脑中动脉缺血2h再灌注模型。60只健康雄性SD大鼠随机分为4组:正常对照组(6只),假手术组(6只),缺血对照组(24只)和移植治疗组(24只);后2组又分为再灌注7d、14d、21d、28d组(各6只)。体外培养脂肪基质细胞,诱导分化为神经干细胞。造模成功后24h,移植治疗组经尾静脉移植人脂肪组织来源的神经干细胞悬液(细胞浓度为2×106/ml),缺血对照组经尾静脉注射生理盐水,假手术组不做任何处理。TUNEL法检测细胞凋亡,免疫组化SABC法检测Bcl-2、Bax表达。结果与缺血对照组比较,移植治疗组各时间点的细胞凋亡数均明显减少(均P<0.01),Bcl-2阳性细胞数明显增高(均P<0.01),Bax阳性细胞数明显减少(P<0.05～0.01)。结论人脂肪组织来源的神经干细胞可能通过上调Bcl-2蛋白表达、下调Bax蛋白表达,减少局灶性脑缺血细胞凋亡;对脑缺血再灌注损伤后的神经细胞起保护作用。Objective To investigate the effects of transplantation of neural stem cells originated from human adipose tissue on the cell apoptosis and the expression of protein Bcl-2 and Bax following focal cerebral ischemia-reperfusion（CIR）in rats.Methods The models of middle cerebral artery occlusion（MCAO）2 h and reperfusion of rats were made by artery thread embolism method.60 healthy male Sprague-Dawley rats were randomly divided into 4 groups：normal control group（n=6）,sham-operated group（n=6）,ischemia control group（n=24）and transplantation group（n=24）.Ischemia control group was divided into CIR 7 d,14 d,21 d and 28 d groups（each group had 6 rats）,and so did the transplantation group.Adipose tissue-derived stromal cells were cultured and induced into neural stem cells in vitro.Transplantation group was transplanted neural stem cells（with concentration of 2×106/ml）through tail vein at 24 h of CIR.Ischemia control group was injected with isodose normal saline through the same vein.The cell apoptosis was detected by TUNEL staining and the expression of protein Bcl-2 and Bax were deteted by immunohistochemical staining.Results Compared with the ischemia control group,the number of apoptotic cells in transplantation group decreased significantly at every time point（all P〈0.01）,the number of Bcl-2 protein positive cells was increased（all P〈0.01）,and the number of Bax protein positive cells was decreased（P〈0.05～0.01）.Conclusions The transplantation of neural stem cells originated from human adipose tissue can decrease the number of apoptotic cells after CIR by up regulation of Bcl-2 and down regulation of Bax.It can protect the neurocyte after CIR.

关 键 词：脂肪基质细胞 神经干细胞 脑缺血再灌注 细胞凋亡 Bcl-2 Bax 

分 类 号：R741[医药卫生—神经病学与精神病学]