Maresin1 Alleviates MSU Crystal-induced Inflammatory Response By Upregulating The Expression Of Prdx5

Objective:Maresin 1 is a recently discovered pro-inflammatory substance with significant anti-inflammatory effects;Prdx5 is a mitochondrial target antioxidant enzyme that can effectively remove ROS in cells and mitochondria.This topic investigated the possible mechanism of Maresin1 alleviating MSU crystal-induced inflammation by regulating the expression of Prdx5,so as to explore the role of Maresin1 in alleviating MSU crystal-induced inflammatory response and its molecular mechanism.Methods:1.Maresin1 pretreated MSU crystal-stimulated BMDMs,and RT-qPCR was used to detect the mRNA expression levels of inflammation-related genes;Western blot was used to detect the protein levels of caspase1 and IL-β in cell lysates and medium supernatants treated with MSU crystals;Flow cytometry was used to detect the total cellular reactive oxygen species(ROS)and mitochondrial ROS levels with DCFH-DA and Mito-SOX fluorescent probes,respectively,and the level of mitochondrial membrane potential with JC-1 probe.2.MSU crystal-stimulated BMDMs and MSU crystal-stimulated BMDMs protein lysates were pretreated with Maresin1 and sent to the company for proteomic detection and related bioinformatics analysis.3.According to the proteomic data,to explore whether Prdx5 regulates the inflammatory response induced by MSU crystals and its molecular mechanism.Purchase Prdx5+/-mice,obtain Prdx5-/-mice after breeding,take the bone marrow of Prdx5-/-and Prdx5+/+ mice,and induce macrophages;BMDMs are transfected with plasmids containing the insertion frame of Prdx5 development;Then,Prdx5 knockout and overexpressed BMDMs were stimulated with MSU crystals,respectively,and the expression levels of inflammation-related genes,total ROS and mitochondrial ROS levels,AMPK phosphorylation levels,and NLRP3 inflammasome activation were detected.4.Prdx5-/--derived BMDMs were treated with Maresin1 to detect the expression levels of inflammation-related genes,total ROS and mitochondrial ROS levels,AMPK phosphorylation levels,and NLRP3 inflammasomes stimulated by MSU crystals after Prdx5 knockout by Maresin1 activation effect.5.To construct mouse models of peritonitis and arthritis,to explore in vivo Maresin1 alleviating the inflammatory response induced by MSU crystals and the protective effect of Maresin1 on the inflammatory response induced by MSU crystals in Prdx5-/mice.Results:MSU crystals pretreated with Maresin1 reduced the expression of inflammatory genes,caspase1 and IL-β in BMDMs cells.The related indicators of mitochondrial function indicated that Maresin1 alleviated the mitochondrial dysfunction induced by MSU crystals.Prdx5 effectively alleviated the decrease in cellular Prdx5 expression caused by MSU crystals.In Prdx5-/-BMDMs cells,the expression of inflammation-related genes was increased,caspase1 and IL-β protein levels were increased,and mitochondrial dysfunction was more severe,while the opposite results were seen in Prdx5-overexpressing BMDMs cells.Maresin1 increases the expression of Prdx5.Regulating the expression of Prdx5 reduces the expression of related inflammatory genes,reduces the levels of caspase1 and IL-β proteins,affects the phosphorylation of AMPK,maintains the balance of TRX1 and TXNIP protein levels,and alleviates MSU crystal-induced peritonitis in mice and MSU crystal-induced arthritis in Prdx5-/-mice.Conclusion:Maresin1 alleviates the expression of MSU crystal-induced inflammation-related genes and the activation of NLRP3 inflammasome by regulating the expression of Prdx5,providing a new theoretical basis and therapeutic target for the treatment of gouty arthritis in the future.