刘流, 刘琳, 梁力川, 等.IL-1β预处理脂肪间充质干细胞促进VEGF蛋白分泌和巨噬细胞M2型极化的研究.四川大学学报(医学版),2019,50(2):171-176
IL-1β预处理脂肪间充质干细胞促进VEGF蛋白分泌和巨噬细胞M2型极化的研究
Effects of IL-1β Pretreated Adipose Tissue-derived Mesenchymal Stem Cells on VEGF Secretion and Macrophage M2 Polarization
  
中文关键词:  间充质干细胞 U937细胞 巨噬细胞极化前列腺素E2血管内皮生长因子
英文关键词:Mesenchymal stem cellsU937 cellsMacrophage polarizationProstaglandin E2Vascular endothelial growth factor
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中文摘要:
      目的探讨白细胞介素(IL)-1β预处理脂肪间充质干细胞(ADMSCs)对血管内皮生长因子(VEGF)蛋白分泌和巨噬细胞M2型极化的影响及作用机制。方法IL-1β预处理ADMSCs 24 h,Western blot法检测ADMSCs中环氧合酶-2(COX-2)蛋白表达,ELISA法测定ADMSCs培养基中前列腺素E2(PGE2)和VEGF蛋白质量浓度; IL-1β预处理的ADMSCs细胞培养基培养丙二醇甲醚醋酸酯(PMA)-U937巨噬细胞(M0巨噬细胞)72 h后,实时荧光定量PCR法检测PMA-U937细胞CD163(巨噬细胞M2型极化标志物)和IL-10 mRNA表达。慢病毒干扰ADMSCs细胞中COX-2表达后,观察ADMSCs培养基中VEGF蛋白质量浓度(ELISA法)和PMA-U937巨噬细胞中CD163和IL-10 mRNA的表达(实时荧光定量PCR法)。结果IL-1β预处理ADMSCs细胞后上调其COX-2蛋白表达,增加PGE2和VEGF蛋白分泌(P<0.05);IL-1β预处理的ADMSCs细胞培养基可上调PMA-U937巨噬细胞中CD163和IL-10 mRNA表达水平(P<0.01)。干扰ADMSCs细胞中COX-2蛋白表达后,可抑制IL-1β预处理的ADMSCs细胞中VEGF蛋白分泌(P<0.05),亦可抑制其对PMA-U937巨噬细胞中CD163和IL-10 mRNA表达的影响(P<0.05)。结论IL-1β预处理ADMSCs细胞可通过COX-2-PGE2信号轴,增加VEGF蛋白分泌,以及诱导PMA-U937巨噬细胞M2型极化,提示IL-1β预处理能够增强ADMSCs细胞抗炎活性。
英文摘要:
      ObjectiveTo investigate the effects of interleukin (IL)-1β preteated adipose tissue-derived mesenchymal stem cells (ADMSCs) on vascular endothelial growth factor (VEGF) secretion and macrophages M2 polarization. MethodsAfter IL-1β pretreated ADMSC for 24 h, the expression of cyclooxygenase-2 (COX-2) was detected using Western blot, and the secretions of prostaglandin E2 (PGE2) and VEGF were measured by ELISA method. Conditioned media collected from ADMSCs was used for culturing PMA-U937 macrophages for 72 h, and the expressions of CD163 and IL-10 mRNAs were detected using qRT-PCR. After inhibition of COX-2 expression in ADMSCs by Lentivirus silencing, secretion of VEGF and CD163 and IL-10 mRNA expressions were detected. ResultsIL-1β pretreating ADMSCs increased the expression level of COX-2, enhanced PGE2 and VEGF secretions (P<0.05). In addition, the mRNA levels of CD163 and IL-10 were upregulated when ADMSCs were pretreated by IL-1β in PMA-U937 cells. After downregulation of COX-2 in ADMSCs, the secretion of VEGF was suppressed, and the mRNA levels of CD163 and IL-10 were also significantly down-regulated. ConclusionIL-1β pretreatment enhanced the secretion of PGE2 and VEGF, and induced macrophage M2 polarization, which depended on COX-2-PGE2 signal pathway.
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