Abstract:
Depending on local cues, macrophages can polarize into classically activated (M1) or alternatively activated (M2) phenotypes. This study investigates the impact of polarized macrophage-derived Extracellular Vesicles (EVs) (M1 and M2) and their cargo of miRNA-19a-3p and miRNA-425-5p on TGF-β production in lung fibroblasts. EVs were isolated from supernatants of M0, M1, and M2 macrophages and quantified using nanoscale flow cytometry prior to fibroblast stimulation. The concentration of TGF-β in fibroblast supernatants was measured using ELISA assays. The expression levels of miRNA-19a-3p and miRNA-425-5p were assessed via TaqMan-qPCR. TGF-β production after stimulation with M0-derived EVs and with M1-derived EVs increased significantly compared to untreated fibroblasts. miRNA-425-5p, but not miRNA-19a-3p, was significantly upregulated in M2-derived EVs compared to M0- and M1-derived EVs. This study demonstrates that EVs derived from both M0 and M1 polarized macrophages induce the production of TGF-β in fibroblasts, with potential regulation by miRNA-425-5p.
摘要:
根据当地的线索,巨噬细胞可以极化为经典激活的(M1)或激活的(M2)表型。这项研究调查了极化的巨噬细胞衍生的细胞外囊泡(EV)(M1和M2)及其miRNA-19a-3p和miRNA-425-5p的货物对肺成纤维细胞中TGF-β产生的影响。从M0、M1和M2巨噬细胞的上清液中分离EV,并在成纤维细胞刺激之前使用纳米级流式细胞术定量。使用ELISA测定法测量成纤维细胞上清液中TGF-β的浓度。通过TaqMan-qPCR评估miRNA-19a-3p和miRNA-425-5p的表达水平。与未处理的成纤维细胞相比,用M0衍生的EV和M1衍生的EV刺激后的TGF-β产生显着增加。miRNA-425-5p,但不是miRNA-19a-3p,与MO和M1衍生的EV相比,M2衍生的EV显着上调。这项研究表明,来自M0和M1极化巨噬细胞的EV诱导成纤维细胞中TGF-β的产生,与miRNA-425-5p的潜在调控。