Role of Pyrin in macrophage M1 polarization

Abstract:

Objective：To investigate the role of Pyrin in macrophage M1 polarization. Methods：THP-1 cells were cultured and periph－eral blood mononuclear cell（PBMC），bone marrow-derived macrophage（BMDM） were extracted. These three kinds of cells were ran－domly divided into control group and treatment group. Treatment group was treated with lipopolysaccharide（LPS，100 ng/mL） and in－terferon-γ（IFN-γ，20 ng/mL） to construct macrophage M1 polarization models（THP-1 for 18 h，PBMC and BMDM for 24 h）. qRT-PCR，Western blot and flow cytometry（FCM） were used to test the markers of M1 polarization，qRT-PCR and Western blot were used to test the expression of MEFV and Pyrin. Results：Compared with those of control group，qRT-PCR detection of THP-1-M1 polarization markers：TNF-α（t=-4.360，P=0.007），IL-1β（t=-8.329，P=0.000），IL-6（t=-2.924，P=0.033），CD14（t=-2.858，P=0.035），CD80（t=-3.433，P=0.019） and Western blot detection of THP-1-M1 polarization markers：TNF-α（t= -3.827，P=0.031），IL-1β（t=-4.189，P=0.025），IL-6（t=-5.345，P=0.002） were significantly increased after the treatment with LPS and IFN-γ. The mRNA levels of TNF-α（t=-2.893，P=0.034），IL-1β（t=-3.606，P=0.015），IL-6（t=-2.895，P=0.034），CD14（t=-2.645，P=0.046），CD80（t=-3.648，P=0.015） of PBMC-M1 polarization markers and the mRNA levels of TNF-α（t=-6.123，P=0.002），IL-1β（t=-2.697，P=0.043），MCP-1（t=-4.335，P=0.007），iNOS（t=-3.607，P=0.015） of BMDM-M1 polarization markers were signifi－cant higher after the treatment with LPS and IFN-γ. Treatment group displayed higher positive cells of THP-1-M1 polarization marker HLA-DR（t=-3.270，P=0.017） and BMDM-M1 polarization marker F4/80+CD11c+（t=-3.833，P=0.009） than control group. These results suggested that the M1 polarization models were constructed successfully. In these three M1 polarization models，qRT-PCR and Western blot detection of MEFV（t=-3.226，P=0.023） and Pyrin（t=-8.591，P=0.000） of THP-1-M1 macrophages were significantly increased. The mRNA level of MEFV of PBMC-M1 macrphages（t=-2.989，P=0.030） and BMDM-M1 macrophages（t=-2.891，P=0.034） were significant higher than control group. Conclusion：The expressions of Pyrin increase in macrophage M1 polariza－tion；Pyrin maybe related to macrophage M1 polarization.