PRDX3 Regulates The Inflammatory Response Induced By MSU Crystals And A Preliminary Exploration Of Its Molecular Mechanism

Objective:Peroxidase 3(PRDX3)has peroxidase activity and it can affect cellular function by regulating mitochondrial reactive oxygen species levels.The activation of NLRP3 inflammatory vesicles and the secretion of IL-1β were associated with the inflammatory response induced by MSU crystals.The main objective of this study was to investigate the role of PRDX3 in gouty arthritis with its possible molecular mechanisms and to explore new ideas for the future treatment of gout.Methods:We collected peripheral blood specimens from outpatients with gout and healthy individuals from the physical examination center of the Affiliated Hospital of North Sichuan Medical College during 2020.3-2021.11,recorded patients’ age,BMI and laboratory indexes such as blood uric acid,sedimentation,C-reactive protein,leukocytes and platelets.By isolating serum and peripheral blood mononuclear cells(PBMCs),the expression levels of serum IL-1β were detected by ELISA and the mRNA expression levels of PRDX3 were detected by RT-qPCR to compare the expression differences between normal subjects and gout patients in the acute and intermittent phases.Mouse bone marrow cells were isolated and cultured by M-CSF stimulation to obtain mouse bone marrow-derived macrophages(BMDMs),pre-activated using LPS and then stimulated with MSU crystals,and the cells were examined by RT-qPCR and Western Blotting for PRDX3,IL-1β,IL-6,TNF-α,COX-2,iNOS and other related molecules expression levels.Flow cytometric assays were used to detect the levels of total reactive oxygen species(ROS),mitochondrial ROS and mitochondrial membrane potential and analyze their expression differences.Results:The expression levels of PRDX3 were higher in PBMCs of gout patients than in normal subjects,with higher expression levels in patients in the acute phase than in the intermittent phase.Inhibition of PRDX3 expression in BMDMs cells,followed by stimulation with MSU crystals,significantly reduced the expression levels of inflammatory factors such as IL-1β,IL-6,and TNF-α.The expression levels of inflammatory mediators iNOS and COX-2 were also reduced,and mitochondrial reactive oxygen species and total cellular reactive oxygen species were decreased,and the level of mitochondrial membrane potential was increased,alleviating the abnormal mitochondrial function.Overexpression of PRDX3 in BMDMs cells,then stimulated with MSU crystals,significantly increased the expression levels of inflammatory factors and inflammatory mediators,increased mitochondrial reactive oxygen species,total cellular reactive oxygen species,and decreased mitochondrial membrane potential levels,indicating that PRDX3 further promoted abnormal mitochondrial function.Conclusion:PRDX3 may regulate the inflammatory response induced by MSU crystals by affecting mitochondrial function;PRDX3 may provide a new therapeutic idea for acute gouty arthritis.