间充质干细胞
CD90型
流式细胞术
免疫印迹
生物
分子生物学
CD11c公司
巨噬细胞极化
细胞生物学
干细胞
CD14型
川地34
脐带
化学
免疫学
巨噬细胞
生物化学
体外
表型
基因
作者
Yongzhao Zhu,Li Li,Zheng Wang,Xipeng Tan,Tao Jin,Ding Lu,Hui Dong,Peng Ye
出处
期刊:PubMed
[National Institutes of Health]
日期:2024-03-01
卷期号:40 (3): 193-198
摘要
Objective To investigate the regulation of IL-1β on the expression of CD200 in human umbilical cord mesenchymal stem cells (hUC-MSCs), its role in macrophage polarization and the underlying mechanism. Methods hUC-MSCs were isolated and cultured in serum-free medium. Morphological observation and the expressions of CD73, CD90, CD105, CD14, CD34, CD45 and HLA-DR were detected by flow cytometry to confirm the properties of mesenchymal stem cells. hUC-MSCs were treated with IL-1β at the final concentration of 20 ng/mL for 24 hours. The proportion of CD200 positive cells was measured by flow cytometry. Real-time quantitative PCR and Western blot analysis were used to detect CD200 mRNA and protein expression levels. hUC-MSCs infected with CD200 overexpression (OE-CD200) and its negative control (OE-NC) lectin virus were treated with IL-1β and co-cultured with PMA-activated THP-1 macrophages. The proportion of CD11c and CD206 positive cells was measured by flow cytometry. hUC-MSCs were treated with IL-1β in combination with PD98059, and the expression of MAPK signaling pathway-related proteins and its effect on CD200 expression were detected by Western blot analysis. Results IL-1β significantly down-regulated the expression of CD200 protein and the proportion of CD200 positive cells. Overexpression of CD200 significantly up-regulated the expression of CD200 in hUC-MSCs, and increased the proportion of CD206-positive macrophages. IL-1β activated the ERK1/2 signaling pathway in hUC-MSCs, and PD98059 up-regulated the expression of CD200 protein in hUC-MSCs treated with IL-1β. Conclusion IL-1β inhibits the expression of CD200 by activating ERK1/2 signaling pathway, and reduces the immunosuppressive effect of hUC-MSCs on regulating the M2-type polarization of macrophages.
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