Expression of IL-1β,TNF-α and NLRP3 in the process of gout inflammation induced by MSU crystals in macrophages with P2X7R overexpression
Objective To observe the expression of NOD-like receptor family 3(NLRP3)protein,interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the process of gout inflammation induced by macrophage monosodium urate(MSU)crystals in macrophages differentiated from leukemia cells overexpressing purinergic receptor P2X ligand-gat-ed ion channel 7(P2X7R).Methods THP-1 cell line were taken and randomly divided into the overexpression group,empty vector group,model group and control group,respectively.Cells in the overexpression group and empty vector group were transfected with P2X7R overexpression plasmid and empty vector plasmid for five days,respectively.THP-1 cells in the vector group,empty vector group and model group were stimulated with 100 ng/mL PMA for three hours and then were differentiated into macrophages.In addition,MSU crystals were dissolved with sodium hydroxide to prepare MSU chyle suspension at a concentration of 100 μg/mL and were incubated in the culture medium for six hours.Cells in the control group were cultured normally.The mRNA and protein of P2X7R in macrophages were measured by RT-PCR and Western blotting,respectively.IL-1β and TNF-α in the supernatant of macrophages were detected by ELISA,and the NLRP3 protein in macrophages was measured by Western blotting.Results Compared with the control group,the rela-tive expression levels of P2X7R mRNA and protein,IL-1β and TNF-α in cell supernatant and NLRP3 protein all increased in the overexpression group,empty vector group and model group(all P<0.05).Compared with the model group and emp-ty vector group,the relative expression levels of P2X7R mRNA and protein,IL-1β and TNF-α in cell supernatant and NL-RP3 protein all increased in the overexpression group(all P<0.05).Conclusion The expression of IL-1β,TNF-α and NLRP3 increases and the levels of IL-1β and TNF-α levels increase in the process of gout inflammation induced by MSU crystals of macrophages differentiated from leukemia cells with P2X7R overexpression,which may be achieved by activat-ing NLRP3 protein.
purinergic receptor P2X ligand-gated ion channel 7 ligandgoutinflammatory factorNod-like recep-tor family 3 inflammasome
国家科技期刊平台
NSTL
万方数据
