脐带来源间充质干细胞抑制巨噬细胞M1极化OA北大核心CSTPCD
Umbilical cord-derived mesenchymal stem cells inhibit macrophage M1 polarization
目的:探究人脐带间充质干细胞(hUC-MSCs)对巨噬细胞M1/M2极化的作用.方法:将hUC-MSCs与佛波酯(PMA)诱导分化的巨噬细胞样细胞(pTHP-1巨噬细胞)共培养后进行转录组测序分析,筛选差异表达基因,进一步进行GO及KEGG富集分析.细胞增殖实验(CCK-8和EdU)分析hUC-MSCs对pTHP-1细胞增殖的影响.流式细胞术检测hUC-MSCs对LPS刺激的pTHP-1巨噬细胞炎症因子TNF-α表达及抑炎因子IL-10表达的影响.qRT-PCR及流式细胞术探究hUC-MSCs对pTHP-1巨噬细胞M1/M2相关分子表型的作用.结果:转录组测序数据分析发现hUC-MSCs与pTHP-1细胞共培养后M1相关基因TNF-α(P<0.05)、HLA-DRA(P<0.01)明显下调,M2相关基因ARG1(P<0.05)明显上调,提示hUC-MSCs抑制巨噬细胞向M1表型极化.GO和KEGG富集分析提示这些表达失调的基因参与调控炎症与免疫应答.hUC-MSCs抑制pTHP-1巨噬细胞增殖,且抑制TNF-α表达(P<0.001),促进IL-10表达(P<0.001).qRT-PCR及流式细胞术分析发现,hUC-MSCs共培养后,pTHP-1细胞HLA-DRA(P<0.05)和CD68(P<0.01)mRNA表达明显下调,且CD14+CD11c+M1型细胞比例下调,而CD163(P<0.001)和CD206(P<0.001)mRNA表达及CD14+CD163+M2型细胞比例明显上调.结论:hUC-MSCs体外抑制巨噬细胞向M1促炎表型极化,诱导向M2抗炎表型极化.
Objective:To explore effect of human umbilical cord mesenchymal stem cells(hUC-MSCs)on macrophage M1/M2 polarization.Methods:hUC-MSCs were co-cultured with pTHP-1 cells which were macrophage-like cells induced by PMA and tran-scriptome sequencing data were analyzed.Differentially expressed genes were screened and analyzed by GO and KEGG enrichment analysis.Effect of hUC-MSCs on pTHP-1 cells proliferation was analyzed by cell proliferation assay(CCK-8 and EdU).Flow cytometry was used to verify influence of hUC-MSCs on relative contents of inflammatory cytokine TNF-α and anti-inflammatory cytokine IL-10 in pTHP-1 cells which were interaction with LPS.Effect of hUC-MSCs on M1/M2-related molecular phenotype of pTHP-1 cells was studied by qRT-PCR and flow cytometry.Results:Transcriptome sequencing data analysis showed that M1-related genes TNF-α(P<0.05)and HLA-DRA(P<0.01)decreased to a great extent and M2-related gene ARG1(P<0.05)increased to a great extent in pTHP-1 cells after co-culture with hUC-MSCs,suggesting that hUC-MSCs inhibited macrophage M1 polarization.GO and KEGG analysis showed that these dysregulated genes regulated inflammation and immune response.hUC-MSCs inhibited proliferation of pTHP-1 cells,reduced content of TNF-α and increased content of IL-10(P<0.001).qRT-PCR and flow cytometry showed mRNA expressions of HLA-DRA(P<0.05)and CD68(P<0.01)and CD14+CD11c+M1 macrophage percentage were down-regulated,while mRNA expressions of CD163(P<0.001),CD206(P<0.001)and CD14+CD163+M2 macrophage percentage were significantly up-regulated in pTHP-1 cells after co-culture with hUC-MSCs.Conclusion:hUC-MSCs inhibit macrophage polarization to M1 and promote polariza-tion to M2 in vitro.
王晓旭;李超然;王惠;杨春娟;刘凤霞;徐栋花
潍坊医学院临床医学院,潍坊 261053潍坊医学院第一附属医院中心实验室,潍坊 261041潍坊医学院第一附属医院过敏反应科,潍坊 261041潍坊医学院第一附属医院中心实验室,潍坊 261041||潍坊医学院第一附属医院风湿免疫科,潍坊 261041
基础医学
脐带间充质干细胞pTHP-1转录组测序巨噬细胞极化
Umbilical cord mesenchymal stem cellspTHP-1Transcriptome sequencingMacrophagePolarization
《中国免疫学杂志》 2024 (004)
673-679 / 7
国家自然科学基金面上项目(82171790);山东省医药卫生科技发展计划(202103100273).
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