人脂肪间充质干细胞通过抑制小胶质细胞迁移活性对MPP^+诱导帕金森细胞模型的保护作用

Protective Effect of Human Adipose-derived Mesenchymal Stem Cells on the MPP^+ Induced Cell Model of Parkinson’s Disease by Inhibiting Microglia Migration

目的本研究旨在探讨经人脂肪间充质干细胞(hADMSCs)诱导后的小胶质细胞(microglia)对多巴胺能神经元的作用。方法利用200 μM MPP ^+处理MN9D细胞建立PD细胞损伤模型,利用hADMSCs条件培养基培养BV2小胶质细胞,建立MPP^+致毒的MN9D细胞与BV2小胶质细胞共培养系统,于Transwell共培养系统检测hADMSCs条件培养基作用下对BV2细胞迁移能力的影响。TUNEL法检测MN9D细胞凋亡,Western blot检测MN9D细胞DAT及TH的表达水平。结果 Transwell结果发现,与MPP +处理组相比,hADMSCs条件培养基处理组(MPP^+/CM)中的BV2细胞迁移率明显减少(40.33±13.19 vs 96.31±10.13)。TUNEL结果发现,经hADMSCs条件培养基培养的BV2小胶质细胞可抑制MN9D细胞凋亡,增加MN9D细胞DAT及TH的蛋白表达水平。结论经hADMSCs条件培养基诱导后的小胶质细胞对DA能神经元起保护作用。

Objective To investigate the effect of microglia pretreated by human adipose-derived mesenchymal stem cells (hADMSCs) on dopaminergic (DA) neurons.Methods The MN9D cells were treated with 200uM 1-methyl-4-phenylpyridinium (MPP ^+) to establish the damage model of Parkinson's Dsease(PD) cells in vitro.The co-culture system of MPP^+ toxic MN9D cells and BV2 microglia cells was established by culturing BV2 microglia cells in the hADMSCs conditioned medium.The Transwell co-culture system was used to detect the effect of the hADMSCs conditioned medium on the migration ability of BV2 microglia cells.The TUNEL method was used to detect the apoptosis of MN9D cells and Western blot was used to determine the expression levels of DAT and TH in MN9D cells.Results The results of Transwell assay revealed that the mobility of BV2 microglia cells in the MPP^+/CM treatment group was significantly reduced when compared with the MPP^+ treatment group (40.33±13.19 vs 96.31±10.13).The TUNEL results showed that the BV2 microglia cells cultured in the hADMSCs conditioned medium inhibited the apoptosis of MN9D cells and increased the expression levels of DAT and TH in MN9D cells.Conclusion BV2 microglia cells induced by the hADMSCs conditioned medium have protective effect on DA neurons.