首页|IL-1β通过激活ERK1/2信号通路抑制人脐带间充质干细胞CD200表达抑制巨噬细胞M2极化

IL-1β通过激活ERK1/2信号通路抑制人脐带间充质干细胞CD200表达抑制巨噬细胞M2极化

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目的 探究白细胞介素1β(IL-1β)调控人脐带间充质干细胞CD200表达及其对巨噬细胞极化的影响及作用机制.方法 无血清培养基分离培养获得人脐带间充质干细胞(hUC-MSC),形态学观察及流式细胞术检测CD73、CD90、CD105、CD14、CD34、CD45、人类白细胞抗原DR(HLA-DR)的表达,确定间充质干细胞属性;20 ng/mL IL-1 β处理hUC-MSC 24 h,流式细胞术检测CD200阳性细胞率,实时定量PCR和Western blot法检测CD200 mRNA和蛋白表达水平;佛波酯(PMA)诱导THP-1巨噬细胞活化,并与IL-1β处理感染CD200过表达慢病毒的hUC-MSC共培养,流式细胞术检测CD11c和CD206阳性细胞比例;IL-1β联合细胞外信号调节激酶1/2(ERK1/2)特异性抑制剂PD98059处理hUC-MSC,Western blot法检测细胞丝裂原激活蛋白激酶(MAPK)信号分子与CD200的表达.结果 IL-1β显著下调hUC-MSC CD200蛋白表达与CD200阳性细胞率;过表达CD200显著上调hUC-MSC CD200表达,且CD200过表达hUC-MSC提高巨噬细胞CD206阳性细胞比率;IL-1 β激活hUC-MSC的ERK1/2信号通路,PD98059上调IL-1β处理后hUC-MSC中CD200的蛋白表达.结论 IL-1β通过激活ERK1/2信号通路抑制CD200的表达,进而抑制hUC-MSC对巨噬细胞向M2型极化的促进作用.
IL-1β inhibits macrophage M2 polarization by down-regulating CD200 expression in human umbilical cord mesenchymal stem cells
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.

interleukin-1β(IL-1β)human umbilical cord mesenchymal stem cells(hUC-MSCs)CD200macrophage polarizationextracellular signal-regulated kinase 1/2(ERK1/2)

朱永朝、李莉、王拯、谭希鹏、陶金、丁璐、董辉、叶鹏

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宁夏医科大学总医院医学科学研究院,宁夏银川 750004

白细胞介素1β(IL-1β) 人脐带间充质干细胞(hUC-MSC) CD200 巨噬细胞极化 细胞外信号调节激酶1/2(ERK1/2)

国家自然科学基金宁夏回族自治区重点研发计划宁夏回族自治区重点研发计划宁夏回族自治区自然科学基金

820600202022BEG030882021BEG020322020AAC03387

2024

10.13423/j.cnki.cjcmi.009696

细胞与分子免疫学杂志
中国免疫学会,第四军医大学

细胞与分子免疫学杂志

CSTPCD北大核心
影响因子：0.817
ISSN：1007-8738
年,卷(期)：2024.40(3)
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