Department of Stomatology, Affiliated Hospital of Qingdao University, 16 Jiangsu Road, Qingdao 266003, Shandong, China; School of Stomatology of Qingdao University, Qingdao 266003, China.
Department of Stomatology, Affiliated Hospital of Qingdao University, 16 Jiangsu Road, Qingdao 266003, Shandong, China.
Int Immunopharmacol. 2020 Apr;81:106030. doi: 10.1016/j.intimp.2019.106030. Epub 2019 Dec 1.
Exosomes have been shown to play a strong role in intercellular communication. While GMSCs have been extensively studied, less research exists on exosomes derived from GMSCs, especially on how exosomes affect macrophages. This study aimed to investigate the impact of GMSC-derived exosomes on macrophage polarization and phenotype under inflammatory conditions.
Exosomes were isolated from GMSCs-conditioned media by ultracentrifugation (UC) and characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and western blot (WB). In vitro, GMSC-derived exosomes were co-incubated with macrophages for 24 h in the absence or presence of M1 polarizing conditions in the six-well plate. The protein and mRNA expression levels of M1 and M2 macrophage markers were detected and the supernatants were collected for an enzyme-linked immunosorbent assay (ELISA).
Exosomes were successfully isolated from GMSCs. Macrophages co-cultured with exosomes showed significantly decreased levels of the M1 markers Tumor Necrosis Factor-α (TNF-α), Interleukin-12 (IL-12), CD86 and Interleukin-1β (IL-1β). By contrast, M2 marker Interleukin-10 (IL-10) levels moderately increased. Meanwhile, similar results were acquired in the cell culture supernatants.
GMSC-derived exosomes may promote M1 macrophage transformation into M2 macrophages, reducing the pro-inflammatory factors produced by M1 macrophages.
外泌体在细胞间通讯中发挥着重要作用。虽然已经广泛研究了间充质干细胞(GMSC),但关于源自 GMSC 的外泌体的研究较少,特别是关于外泌体如何影响巨噬细胞的研究。本研究旨在探讨 GMSC 衍生的外泌体在炎症条件下对巨噬细胞极化和表型的影响。
通过超速离心(UC)从 GMSC 条件培养基中分离出外泌体,并通过透射电子显微镜(TEM)、纳米颗粒跟踪分析(NTA)和 Western blot(WB)进行表征。在体外,将 GMSC 衍生的外泌体与巨噬细胞在六孔板中在不存在或存在 M1 极化条件的情况下共孵育 24 小时。检测 M1 和 M2 巨噬细胞标志物的蛋白和 mRNA 表达水平,并收集上清液进行酶联免疫吸附测定(ELISA)。
成功从 GMSC 中分离出外泌体。与外泌体共培养的巨噬细胞中 M1 标志物肿瘤坏死因子-α(TNF-α)、白细胞介素-12(IL-12)、CD86 和白细胞介素-1β(IL-1β)的水平显著降低。相比之下,M2 标志物白细胞介素-10(IL-10)的水平适度增加。同时,在细胞培养上清液中也获得了类似的结果。
GMSC 衍生的外泌体可能促进 M1 巨噬细胞向 M2 巨噬细胞转化,减少 M1 巨噬细胞产生的促炎因子。