Digoxin ameliorates joint inflammatory microenvironment by downregulating synovial macrophage M1-like-polarization and its-derived exosomal miR-146b-5p/Usp3&Sox5 axis.

Relatively low-grade inflammatory of osteoarthritic joints is characterized by synovitis and a catabolic and proinflammatory state of the chondrocytes and plays an important role in osteoarthritis (OA) initiation and exacerbation. Our previous research showed cardiac glycoside compounds might be effective in OA synovitis. However, the effect of digoxin (DIG), an FDA-approved cardenolide, on inflammation inhibition of osteoarthritic joints has not been investigated. In the present study, a western blot analysis and immunofluorescence staining revealed that DIG alleviated OA synovitis by inhibiting the M1-like polarization of synovial macrophages in OA patients and collagenase-induced OA (CIOA, with considerable synovitis) mice. Subsequently, the exosomes produced by macrophages and M1-like macrophages treated with or without DIG were isolated and identified. According to miRNA sequencing analysis of these exosomes and subsequent target activity assays, we confirmed DIG controls OA inflammatory microenvironment and promotes chondrogenesis by, at least partly, downregulating the M1-like macrophage-derived exosomal miR-146b-5p/Usp3&Sox5 axis in vitro and in vivo. This research provides reliable experimental evidence supporting the clinical application of DIG as a disease-modifying drug for inflammation-associated OA. Additionally, the spectrum of diseases of inflammation controlled by DIG has been broadened, which prompting research interest in the new function of an "old" FDA-approved drug.