High glucose treated Macrophage exosome characterization and gene expression

To determine whether diabetes affects macrophage and contributes to fibrosis through paracrine mechanism, we evaluated the effect of macrophage-exosome on inflammation and fibrogenesis response, <i>in vitro</i>. Exosomes were isolated from conditioned media of macrophages exposed 48hrs to HG or NG conditions (Exo-HG or Exo-NG), transferred to fibroblasts (NIH3/T3 cells) and after 24hrs, inflammatory and fibrogenesis response was assessed. Western blot analysis shows expression of exosome markers such as HSP70, Annexin V, Flot1 etc. To demonstrate the effect of macrophage exosome on target cell function, we incubated fibroblast cells with macrophage-exosome for 24h. qRT-PCR data shows that fibroblast co-cultured with exosomes from HG-treated macrophages (Exo-HG) significantly upregulates mRNA expression of several inflammatory and fibrogenesis signaling molecules such as IL-1β, TGF-β, MMP9 and Col1A1.<br>