Exosomes Derived From IL-1b-primed MSCs Inhibit IL-1β And TNF-α-mediated Inflammatory Response In SW982 Cells

초록

Purpose The present study investigated the anti-inflammatory effect and mechanism of exosomes derived from IL-1β-primed MSCs in IL-1β and TNF-α treated SW982 human synovial cells. Methods and Materials Exosomes were isolated using Exo quick from the supernatant of un-primed or IL-1b-primed MSCs at 15-20 ug/ ml (MSC-Exo vs IL-1b MSC-Exo). The size and morphology of exosomes were characterized by nanoparticle tracking analysis (NTA) and scanning electron microscope (SEM), respectively. Markers of exosome were analyzed by Western blotting. SW982 cells were treated with IL-β (10 ng/ ml) and TNF-α (25 ng/ ml) as an synovial inflammatory model in the presence of MSCs-Exo or IL-β MSCs-Exo for 24h. The expression of inflammation-related genes of IL-1, IL-6 and MCP-1 was examined by real-time RT-PCR. miRNA levels in cells and in isolated exosomes were also examined using real-time RT-PCR. Levels of IkBα and p-IkBα were examined by Western blotting as a down-stream of the inflammatory signal. Results Exosomes isolated from both unprimed and IL-1b-primed MSCs showed round morphology and approximately 30-300 nm in size. Both of them also expressed CD9, CD63 and CD81 at similar levels in western blot analysis. In the IL-1β and TNF-α induced inflammatory model in SW982 cells, IL-1β MSC-Exo showed higher ability than MSCs-Exo to inhibit expression of IL-1β, IL-6 and MCP-1. miRNA 147b was identified at higher levels in IL-1β MSC-Exo than in MSC-Exo among anti-inflammatory miRNAs examined. The amount of IκBα was higher in IL-1β and TNF-α-treated SW982 in the presence of IL-1β MSC-Exo, implicating its role in the anti-inflammatory pathway of miRNA 147b.