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 hPRDX5

关键词:  重组人过氧化物还原酶-5  人脐带来源间充质干细胞  神经干细胞样细胞  神经元样细胞  Toll样受体4

Key words:  recombinant human peroxiredoxin-5  human umbilical cord mesenchymal stem cells  neural stem cell-like cells  neuronal-like cells  toll-like receptor 4

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 peroxidase-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； 10 μg· mL-1 hPRDX5；阳性对照组（N2、B27、bFGF和EGF）；阳性对照+ 1 μg· mL-1 hPRDX5，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染色和阿利辛蓝染色均呈阳性。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 Exploring the feasibility and possible mechanisms of differentiation of human umbilical cord mesenchymal stem cells into neural stem cell-like cells induced by recombinant hPRDX5. METHODS Take normal full-term neonatal umbilical cord under sterile conditions, culture and identify hUC MSCs. Divide hUC-MSCs cultured until the 4th generation 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-induced hUC-MSCs differentiation into NSC-like cells. Observe cell morphology through an inverted microscope. Immunohistochemical detection of NSCs marker nestin. Immunofluorescence staining was used to detect microtubule associated protein 2 in neurons. Western blotting was used to detect Nestin and toll-like receptor 4, a signaling pathway associated protein. RESULTS HUC-MSCs express high levels of CD29, CD73, CD90, and CD105, but do not express CD19, CD34, CD45, and HLA-DR. Osteogenesis, adipogenesis, and chondrogenesis induced differentiation were all positive for Alizarin Red staining, Oil Red O staining, and Alisin Blue staining. hPRDX5 induced NSC-like cells surface marker Nestin positivity. hPRDX5 induced differentiation of NSC-like cells into MAP2 positive neuronal-like cells. The expression of signaling pathway related protein TLR4 in NSC-like cells is elevated. 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.