人促红细胞生成素基因修饰人羊膜间充质干细胞蛛网膜下腔移植对脑损伤的影响  

Human erythropoietin-modified human amniotic mesenchymal stem cells via subarachnoid transplantation promote neurologic recovery from brain injury

作　　者：吕刚[1,2] 张宏远[2] 

机构地区：[1]天津医科大学研究生院,天津市300350 [2]天津市津南区咸水沽医院脑系科,天津市300350

出　　处：《中国组织工程研究》2016年第23期3432-3438,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:有研究表明促红细胞生成素是一种调节骨髓造血功能的糖蛋白,不仅具有调节中枢神经系统发育作用,还可以营养神经及保护神经,但实验结果证实促红细胞生成素基因修饰人羊膜间充质干细胞移植成为治疗脑损伤的新途径。目的:观察促红细胞生成素修饰的人羊膜间充质干细胞蛛网膜下腔移植对脑损伤大鼠神经损伤后功能恢复的影响。方法:体外培养人羊膜间充质干细胞,构建真核表达质粒pcD NA3.1促红细胞生成素,并转染入人羊膜间充质干细胞。用Western blot检测人羊膜间充质干细胞在转染前后人促红细胞生成素蛋白的表达。构建大脑中动脉闭塞模型大鼠,用促红细胞生成素修饰的人羊膜间充质干细胞尾静脉移植,并设模型组和人羊膜间充质干细胞组作对比。结果与结论:(1)转染结果鉴定:Western blot结果显示,转染人促红细胞生成素基因的人羊膜间充质干细胞体外能表达人促红细胞生成素;(2)行为学变化、生长相关蛋白43及水通道蛋白9表达:神经功能评分,RT-PCR和Western blot检测结果显示,人促红细胞生成素基因+人羊膜间充质干细胞组改良的神经功能缺损评分、脑组织生长相关蛋白43及水通道蛋白9基因及蛋白的表达明显小于其他2组(P<0.05);(3)细胞存活情况:人促红细胞生成素+人羊膜间充质干细胞组CM-Dil的阳性细胞数最多(P<0.05),人羊膜间充质干细胞组次之(P<0.05),脑损伤组最少(P<0.05)。(4)结果证实,促红细胞生成素基因修饰人羊膜间充质干细胞移植可降低脑损伤区周围生长相关蛋白43及水通道蛋白9基因及蛋白的表达,抑制神经细胞的凋亡,促进神经功能恢复。BACKGROUND： Studies have shown that as a regulator of bone marrow function erythropoietin is a glycoprotein that controls the development of the central nervous system and has neurotrophic and neuroprotective potential. Therefore, transplantation of human amniotic mesenchymal stem cells genetically modified by human erythropoietin is a new choice for brain injury treatment. OBJECTIVE： To observe the effect of transplantation of human amniotic mesenchymal stem cells genetically modified by human erythropoietin on the functional recovery from brain injury in rats. METHODS： Eukaryotic expression plasmid pcD NA3.1 carrying erythropoietin was successfully constructed and transferred into amniotic mesenchymal stem cells cultured in vitro. Expression of erythropoietin was detected using western blot assay before and after transfection. Rat models of middle cerebral arterial occlusion was made and given transplantation of transfected amniotic mesenchymal stem cells via the tail vein（transfection group）. Additionally, model and simple cell transplantation groups were set in a comparative study. RESULTS AND CONCLUSION： Findings from western blot detection showed that transfected cells could express human erythropoietin. Compared with the other groups, modified neurologic severity scores, growth-associated protein 43 and aquaporin 9 at m RNA and protein levels were all decreased significantly in the transfection group. Furthermore, the number of cells positive for CM-Dil was highest in the transfection group, followed by simple cell transplantation group, and lowest in the model group（all P 0.05）. Overall findings from this study show that human erythropoietin-modified human amniotic mesenchymal stem cell transplantation promotes neurologic recovery from brain injury through eliciting a reduction in growth-associated protein 43 and aquaporin 9 at m RNA and protein levels as well as inhibiting cell apoptosis.

关 键 词：干细胞 组织工程 脑损伤 移植 人促红细胞生成素 干细胞移植 人羊膜间充质干细胞 大鼠 生长相关蛋白43 水通道蛋白9 基因 修饰 神经功能 

分 类 号：R394.2[医药卫生—医学遗传学]