脂肪组织来源的神经干细胞移植对大鼠局灶性脑缺血细胞凋亡及Caspase-3蛋白表达的影响  

Effects of neural stem cells from human adipose tissue-derived stromal cellstransplantation on apoptosis and caspase-3 after focal cerebral ischemia in rats

作　　者：刘斌[1] 王亚薇[1] 董静[1] 王瑞敏[2] 张晋霞[1] 

机构地区：[1]河北联合大学附属医院神经内一科,河北唐山063000 [2]河北联合大学实验中心,河北唐山063000

出　　处：《中国现代医学杂志》2011年第19期2231-2235,共5页China Journal of Modern Medicine

基　　金：河北省自然科学基金(No:C2008000994);河北省优秀专家出国培训基金(No:2007);唐山市科研基金(No: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细胞凋亡及Caspase-3蛋白表达的影响。方法线栓法制作大鼠大脑中动脉缺血(MCAO)2 h再灌注模型。60只健康雄性SD大鼠随机分为4组:正常组(6只),假手术组(6只),缺血对照组(24只)和移植治疗组(24只)。后两组又分为缺血2 h再灌7、14、21和28 d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Caspase-3表达。结果与缺血对照组比较,移植治疗组各时间点(7、14、21和28 d)的细胞凋亡数均明显减少(均P<0.01),Caspase-3蛋白表达均明显减少(均P<0.01)。结论脂肪组织来源的神经干细胞可能通过下调Caspase-3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 study the effects of neural stem cells from human adipose tissue-derived stromal cells transplantation on the apoptosis and Caspase-3 after focal cerebral ischemia in rats.【Methods】 The middle cerebral artery occlusion（MCAO）2 h and reperfusion models were made by intraluminal vascular occlusion in rats.60 healthy male Sprague-Dawley rats were separated into 4 groups randomly： normal group（n =6）,sham-operated group（n =6）,ischemia control group（n =24） and transplantation treated group（n =24）.Ischemia control group and transplantation treated group were divided randomly into 7,14,21 and 28 d reperfusion group（n =6）.Adipose tissue-derived stromal cells were cultivated,proliferated and induced into neural stem cells in vitro.Transplantation treated group was transplanted neural stem cells（2×106/mL） through tail vein at 24 h hours after MACO.Equivalent volume of normal saline was injected into the rats in ischemia control group.The apoptotic cell was detected by TUNEL staining and the expression of Caspase-3 protein were detected by immunohistochemical SABC staining.【Results】 Compared with the ischemia control group,the number of apoptotic cells in the transplantation treated group decreased at every time point（7 d,14 d,21 d,28 d） after transplantation（all P 0.01）,the Caspase-3 protein cells in the transplantation treated group were significantly reduced.（all P 0.01）.【Conclusion】 Neural stem cells differentiated from human adipose tissue-derived stromal cells can decrease the number of apoptotic cells after cerebral ischemia and protect the nerve cells from cerebral ischemia-reperfusion injury by down-regulation Caspase-3.

关 键 词：脂肪基质细胞 神经干细胞 脑缺血 细胞凋亡 胱冬酶-3 

分 类 号：R-332[医药卫生]