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

发布时间：2018-02-09 22:37

本文关键词： 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水通道蛋白基因修饰神经功能　出处：《中国组织工程研究》2016年23期 　论文类型：期刊论文

【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(P0.05);(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(P0.05),人羊膜间充质干细胞组次之(P0.05),脑损伤组最少(P0.05)。(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。
[Abstract]:Background: erythropoietin is a glycoprotein that regulates hematopoietic function in bone marrow. It not only regulates the development of the central nervous system, but also nourishes and protects the nerve. However, the experimental results confirmed that the transplantation of human amniotic mesenchymal stem cells modified with erythropoietin gene was a new way to treat brain injury. Objective: to observe the subarachnoid space of erythropoietin modified human amniotic mesenchymal stem cells. Effects of transplantation on functional recovery after nerve injury in rats with brain injury methods: human amniotic mesenchymal stem cells were cultured in vitro. The eukaryotic expression plasmid pcD NA3.1 erythropoietin was constructed. Western blot was used to detect the expression of human erythropoietin protein before and after transfection of human amniotic mesenchymal stem cells. Erythropoietin modified human amniotic mesenchymal stem cells were transplanted into caudal vein and compared with model group and human amniotic mesenchymal stem cell group. Human erythropoietin gene transfected human amniotic mesenchymal stem cells could express human erythropoietin (EPO) 2) in vitro. The expression of growth-associated protein 43 and aquaporin 9 were detected by RT-PCR and Western blot. Modified neurological impairment score of human erythropoietin gene human amniotic mesenchymal stem cells group, The expression of brain growth-associated protein 43 and aquaporin 9 gene and protein were significantly lower than those of other two groups (P 0.05). The number of CM-Dil positive cells in human erythropoietin human amniotic mesenchymal stem cells group was the highest (P 0.05). Human amniotic mesenchymal stem cells (HMMSCs) group was followed by P0.05, and brain injury group was the least (P0.05N. 4). Transplantation of human amniotic mesenchymal stem cells modified with erythropoietin gene can reduce the expression of growth-associated protein 43 and aquaporin-9 gene and protein around the brain injury area, inhibit the apoptosis of nerve cells and promote the recovery of nerve function.
【作者单位】：天津医科大学研究生院;天津市津南区咸水沽医院脑系科;
【分类号】：R651.15

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