| Objective:Previous studies have reported that HSP60 can regulate the TLR4-MyD88-NF-κB signaling pathway and the activation of NLRP3 inflammasome.The main purpose of this study was to explore the role and molecular mechanism of HSP60 in MSU crystal-induced inflammatory response.Methods:Peripheral blood samples from gout patients and healthy people were collected,serum and peripheral blood mononuclear cells were isolated,and the expression level of HSP60 was measured by ELISA and western blotting.After interfering with the expression of HSP60 in THP-1 derived macrophages and RAW264.7 cells,they were stimulated with MSU crystals,the expression level of TLR4-MyD88-NF-κB signaling pathway-related factors and the activation of NLRP3 inflammasome were measured by western blotting and Immunofluorescence.the expression of inflammation-related factors downstream of NF-κB was measured by RT-PCR.The mice were injected intraperitoneally with HSP60 Morpholino(HSP60 Mo)and injected with MSU crystal suspension to induce acute gouty arthritis and acute peritonitis models.The swelling degree of the footpad and ankle joint was measured,and the infiltration of inflammatory cells in the footpad tissue was analyzed,the number of inflammatory cells in the peritoneal lavage fluid was statistically analyzed,and the expression level of related proteins in the footpad tissue was measured by western blotting.Results:The expression level of HSP60 in PBMCs andserum of gout patients was higher than that of normal people,while the expression level of HSP60 in PBMCs andserum of patients with acute phase was higher than that of non-acute phase.Different concentrations of MSU crystals stimulated THP-1 derived macrophages and RAW264.7 cells,The expression level of HSP60 almost increased in a dose-dependent manner.It interferes with the expression of HSP60 in THP-1 derived macrophages and RAW264.7 cells,and inhibited the activation of the TLR4-MyD88-NF-κB signaling pathway and the activation of NLRP3 inflammasome by MSU crystals.Suppresses HSP60 expression in mice,MSU crystal-induced acute gouty arthritis and acute peritonitis mouse models reduced swelling coefficients of footpad and ankle joint,HE staining of footpad tissue sections shows reduced inflammatory cell infiltration,andthe number of inflammatory cells was reduced.Conclusion:HSP60 may participate in the MSU-induced inflammatory response by promoting the TLR4-NF-κB signaling pathway and activating the NLRP3 inflammasome.HSP60 may be a potential target for the treatment of acute gouty arthritis. |
