Role Of Pyrin In Macrophage M1 Polarization

ObjectivePyrin is a functional protein encoded by the MEFV gene and is mainly expressed in polymorphonuclear leukocytes and monocytes,its mutation is the cause of many inflammatory related diseases.Macrophage polarization is the different functional status of macrophages in different environments.Macrophage M1 polarization exhibits strong pro-inflammatory capabilities.Since Pyrin and macrophage M1 polarization play an important role in innate immunity,it is of great significance to study whether Pyrin can affect macrophage polarization.This study aims to investigate the role of Pyrin in macrophage M1 polarization.MethodsTHP-1 cells were cultured and peripheral blood mononuclear cell(PBMC),bone marrow-derived macrophage(BMDM)were extracted.These three kinds of cells were treated with lipopolysaccharide(LPS,100 ng/mL)and interferon-γ(IFN-y,20 ng/mL)to construct macrophage M1 polarization models(THP-1 for 18 h,PBMC and BMDM for 24 h).qPCR,Western blot and flow cytometry(FCM)were used to test the markers of M1 polarization,qPCR and Western blot were used to test the expression of MEFV and Pyrin.The M1 macrophage model was constructed after the MEFV gene was lowly expressed in the THP-1 cells using siRNA,and the markers of M1 polarization were detected by qPCR.ResultsCompared with those of control group,qPCR detection of THP-1-M1 polarization markers:TNF-α,IL-1β,IL-6,CD80 and Western blot detection of THP-1-M1 polarization markers:TNF-a,IL-1β,IL-6 were significantly increased after the treatment with LPS and IFN-γ.The mRNA levels of TNF-α,IL-1β,IL-6,CD80 of PBMC-M1 polarization markers and the mRNA levels of TNF-α,IL-1β,MCP-1,iNOS of BMDM-M1 polarization markers were significant higher after the treatment with LPS and IFN-γ.Polarization group displayed higher positive cells of THP-1-M1 polarization marker HLA-DR and BMDM-M1 polarization marker F4/80+CD11c+ than control group.These results suggested that the M1 polarization models were constructed successfully.In these three M1 polarization models,qPCR and Western blot detection of MEFV and Pyrin of THP-1-M1 macrophages were significantly increased.The mRNA level of MEFV of PBMC-M1 macrphages and BMDM-M1 macrophages were significant higher than control group.THP-1 cells with low expression MEFV induced M1 polarization,and qPCR detection of markers of M1 polarization:IL-1β,IL-6 and HLA-DR had no significant changes compared with the control group.ConclusionThe expression of Pyrin increase during macrophage M1 polarization,Pyrin maybe related to macrophage M1 polarization.