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Objective To explore the effect of ulinastatin on the pyroptosis of macrophages induced by lipopdysaccharides (LPS)/nigericin. Methods Bone marrows (BM) were isolated from the mice, and then induced into macrophages using macrophage colony-stimulating factor (MCSF). BM macrophages were pretreatment with 1000U/ml ulinastatin. Then, macrophages were primed with 100ng/ml LPS for 4 hours, followed by 10μmol/L nigericin treatment for 2 hours before the analysis. Cells without ulinastatin treatment or without LPS/nigericin prime were used as control. The cell viability was measured by MTT and LDH release rate was also measured using spectrophotometry. The alterations of caspase-1 and Gasdermin D protein (GSDMD) expression and the mRNA of interleukin (IL)-1β, IL-18 were detected using Western blotting and RT-qPCR, respectively. The concentrations of IL-1β, IL-18, TNF-α, and IL-6 were measured by ELISA. Results LPS combined with nigericin were used to establish the cell pyroptosis model in vitro. Our results showed that, ulinastatin pretreatment increased the survival rate of macrophage after LPS/nigericin stimuli (LPS/nigericin+PBS group vs. LPS/nigericin+UTI group, 73.40% vs. 86.30%, P |
