重组人过氧化物还原酶-5诱导人脐带间充质干细胞向神经干细胞样细胞分化

    Differentiation of Human Umbilical Cord Mesenchymal Stem Cells into Neural Stem Cell-like Cells Induced by Recombinant Human Peroxiredoxin-5

    • 摘要:
      目的  探讨人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUC-MSCs)在重组人过氧化物还原酶-5(human peroxiredoxin-5,hPRDX5)诱导下分化为神经干细胞(neural stem cells,NSCs)样细胞的可行性以及可能的作用机制。
      方法 通过无菌条件取正常足月新生儿脐带,培养和鉴定hUC-MSCs。将培养至第4代的hUC-MSCs分为5组:空白对照组、1 µg·mL−1 hPRDX5组、10 µg·mL−1 hPRDX5组、阳性对照组(N2、B27、bFGF和EGF)、阳性对照+1 µg·mL−1 hPRDX5组,考察hPRDX5诱导hUC-MSCs分化成NSCs样细胞的潜能。通过倒置显微镜观察细胞形态。免疫组织化学检测NSCs标志物巢蛋白(Nestin)。免疫荧光染色检测神经元微管相关蛋白2(microtubule-associated protein 2,MAP2)。Western blotting检测Nestin和信号通路相关蛋白Toll样受体4(toll-like receptor 4,TLR4)。
      结果 hUC-MSCs高表达CD29、CD73、CD90和CD105,不表达CD19、CD34、CD45和HLA-DR。成骨、成脂和成软骨诱导分化后经茜素红染色、油红O染色和阿利新蓝染色均呈阳性。hPRDX5诱导的NSCs样细胞表面标志物Nestin阳性。hPRDX5诱导的NSCs样细胞分化为MAP2阳性的神经元样细胞。NSCs样细胞的信号通路相关蛋白TLR4表达升高。
      结论 hPRDX5能促进hUC-MSCs向NSCs样细胞分化,潜在机制可能是通过TLR4信号通路实现的。

       

      Abstract:
      OBJECTIVE  To investigate the feasibility and possible mechanism of human umbilical cord mesenchymal stem cells(hUC-MSCs) differentiation into neural stem cell(NSC)-like cells induced by recombinant human peroxiredoxin-5(hPRDX5).
      METHODS  Umbilical cords from normal full-term neonates were obtained under sterile conditions, and hUC-MSCs were cultured and identified. hUC-MSCs at passage 4 were divided into 5 groups: blank control group, 1 µg·mL−1 hPRDX5, 10 µg·mL−1 hPRDX5, positive control group (N2, B27, bFGF, and EGF), positive control+1 µg·mL−1 hPRDX5, to investigate the potential of hPRDX5 in inducing hUC-MSCs differentiation into NSC-like cells. Cell morphology was observed using an inverted microscope. Immunohistochemistry was performed to detect the NSC marker Nestin. Immunofluorescence staining was used to detect microtubule-associated protein 2 (MAP2). Western blotting was employed to detect the expression of Nestin and toll-like receptor 4 (TLR4), a signaling pathway-related protein.
      RESULTS  hUC-MSCs expressed high levels of CD29, CD73, CD90, and CD105, but did not express CD19, CD34, CD45, or HLA-DR. Following osteogenic, adipogenic, and chondrogenic differentiation, positive staining was observed with Alizarin red, oil red O, and Alcian blue, respectively. NSC-like cells induced by hPRDX5 showed positive immunostaining for Nestin. Furthermore, these hPRDX5-induced NSC-like cells differentiated into MAP2-positive neuron-like cells. The expression of TLR4, a signaling pathway-related protein, was significantly increased in NSC-like cells.
      CONCLUSION  hPRDX5 can promote the differentiation of hUC-MSCs into NSC-like cells, and the underlying mechanism may be achieved through the TLR4 signaling pathway.

       

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