Macrophage pyroptosis promotes synovial fibrosis through the HMGB1/TGF- β1 axis: an in vivo and in vitro study

Macrophages and fibroblasts are the main effector cells in synovial tissue in the knee joint. Our previous studies showed that there was synovial macrophage pyroptosis in knee osteoarthritis (KOA) and that inhibiting this pyroptosis could alleviate synovial fibrosis. In the present study, we aimed to elucidate the mechanism by which macrophage pyroptosis affects synovial fibrosis. We established an LPS/ATP-induced model in macrophages that mimicked the inflammatory environment of KOA and induced macrophage pyroptosis. The TGF-β1, SMAD3, and P-SMAD3, and the synovial fibrosis markers (Collagen I, TIMP1, Vimentin, and TGF-β1) were significantly decreased after fibroblasts were cultured with RAGE inhibitors and SMAD3 inhibitors. Moreover, ELISA and immunofluorescence analysis showed that macrophage pyroptosis induced the release of IL-1β, IL-18, and HMGB1 and caused the translocation of HMGB1 from the fibroblast nucleus to the cell membrane, where it could bind with RAGE. Subsequently, in the synovial tissue of KOA model rats, we observed that inhibiting HMGB1, RAGE, and SMAD3 could alleviate the expression of synovial fibrosis markers (Collagen I, TIMP1, Vimentin, and TGF-β1) at both the mRNA and protein levels. Besides, HE and Sirius Red staining were used to observe the transverse diameter of the right knee. In conclusion, macrophage pyroptosis induced IL-1β, IL-18, and HMGB1, which could be caused HMGB1 to translocate from the fibroblast nucleus and bind with RAGE, activating the TGF-β1/SMAD3 signaling pathway and affecting synovial fibrosis.