【摘 要】
：
目的 探讨神经生长因子(nerve growth factor,NGF)过表达诱导的脐血干细胞(umbilical cord mesenchy-mal stem cells,UMSCs)向多巴胺能神经元分化治疗帕金森病(Parkinson\'s disease,PD)模型大鼠的效果及其对Tribbles同源蛋白3(TRB3)介导的信号通路的影响.方法 建立PD模型,体外培养及诱导UMSCs,将pcDNA-NC、pcDNA-NGF转染至UMSCs中,分为pcDNA-NC组和pcDNA-NGF组.实验设置
【机 构】
：
郑州市第一人民医院神经内科,郑州 450000
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目的 探讨神经生长因子(nerve growth factor,NGF)过表达诱导的脐血干细胞(umbilical cord mesenchy-mal stem cells,UMSCs)向多巴胺能神经元分化治疗帕金森病(Parkinson\'s disease,PD)模型大鼠的效果及其对Tribbles同源蛋白3(TRB3)介导的信号通路的影响.方法 建立PD模型,体外培养及诱导UMSCs,将pcDNA-NC、pcDNA-NGF转染至UMSCs中,分为pcDNA-NC组和pcDNA-NGF组.实验设置4组:NC组(大鼠仅进行手术,不进行注射)、PD组(PD模型)、UMSCs组(正常大鼠中注射含有pcDNA-NGF的UMSCs细胞悬液)、UMSCs+PD组(PD模型大鼠中注射含有pcDNA-NGF的UMSCs细胞悬液),每组各9只.采用实时荧光定量聚合酶链反应(qRT-PCR)检测NGF、神经胶质纤维酸性蛋白(GFAP)、神经元细胞内微管相关蛋白2(MAP2)、微管蛋白(Tubulin)、TRB3 mRNA的表达量;观察各组大鼠旋转圈数;采用免疫组化法检测各组大鼠黑质区酪氨酸羟化酶(TH)阳性细胞数;蛋白免疫印迹法(Western blot)检测NGF、TRB3的蛋白表达量.结果 与pcDNA-NC组比较,pcDNA-NGF组NGF mRNA及蛋白、巢蛋白(Nestin)、GFAP、MAP2、Tubulin mRNA的表达水平升高(P<0.05).与NC组比较,PD组大鼠旋转圈数增多(P<0.05);与UMSCs组比较,UMSCs+PD组大鼠旋转圈数增多(P<0.05);与PD组比较,UMSCs+PD组大鼠旋转圈数减少(P<0.05).与NC组比较,PD组大鼠黑质区TH阳性细胞计数减少(P<0.05);与PD组比较,UMSCs+PD组大鼠黑质区TH阳性细胞计数增多(P<0.05).与NC组比较,PD组大鼠脑组织中TRB3 mRNA及蛋白表达水平升高(P<0.05);与PD组比较,UMSCs+PD组大鼠脑组织中TRB3 mRNA及蛋白表达水平降低(P<0.05).结论 NGF过表达诱导的UMSCs可促进细胞向多巴胺能神经元分化,从而改善PD大鼠的症状,其作用机制可能是通过调控TRB3信号通路而发挥作用.
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