人羊膜间充质细胞的神经生物学特性及其治疗帕金森模型小鼠的实验研究

Neurobiological Characteristics of Human Histo-amniotic Mesenchymal and its Effect to Treat Parkinson's Disease Modle Mice

目的评价人羊膜组织来源间充质细胞(hAMCs)的神经生物学特性及其脑移植对帕金森病(PD)小鼠的治疗观察。方法应用免疫细胞化学染色,通过间充质干细胞、神经干细胞、多巴胺能神经元和神经发生相关标记物Vimentin、STRO-1、nestin、CD133、β-tubulin、TH、DAT、Ngn2和mash-1抗体,鉴定在hAMCs中的表达;RT-PCR检测神经干细胞标记物Vimentin及nestin mRNA在hAMCs的表达。C57BL/6小鼠MPTP(i.p.)建立PD模型,将hAMCs移植到右侧纹状体内。通过自发运动和转杆实验以及抗人线粒体和TH抗体在脑纹状体免疫组织化学形态学,评价hAMCs对PD小鼠的治疗作用。结果经神经细胞培养基诱导培养后的hAMCs表达间充质干细胞、神经干细胞、多巴胺能神经元和上述所有与神经发生相关的特异性标记物。hAMCs移植后的PD小鼠自发运动次数增加(P<0.05),转杆时间延长(P<0.05)。结论hAMCs经体外诱导培养后具有神经细胞特性,移植到PD模型小鼠纹状体内能够改善MPTP所致的运动功能损伤。

Objective To evaluate the neurobiological characteristics of human histio-amniotic mesenchymal （hAMCs） and effect of hAMCs transplantation into the brain to treat Parkinson＇s disease（PD） modle mice.Methods The expressions of mesenchymal stem cells,neural stem cells,dopaminergic neurons and markers related to neurogenesis such as Vimentin,STRO-1,nestin,CD133,β-tubulin,TH,DAT,Ngn2 and mash-1 in hAMCs were evaluated through immunocytochemical stain; and the mRNA transcriptions of neural stem cell markers,Vimentin and nestin in hAMCs were detected by RT-PCR. The PD model was induced by MPTP（i.p.） in C57BL/6 mice transplanted with hAMCs into the right striatum. The therapeutical effect of hAMCs on PD mice was evaluated by spontaneous movement,rotating bar test and the immunohistochemistry of anti-human chondrosome and TH antibodies in striatum.Results hAMCs induced by nerve cells culture medium,expressed mesenchymal stem cells,neural stem cells,dopaminergic neurons and other specific markers related to neurogenesis mentioned above. The frequency of spontaneous movement in PD mice was significantly increased（P〈0.05）,and the time of rotating bar was obviously prolonged（P〈0.05） after transplantation with hAMCs.Conclusion hAMCs possess the characteristics of nerve cells after cultured in vitro and can significantly recover the damage of motor function induced by MPTP after transplantation into striatum in PD model mice.