Objective:To investigate the effect of acacetin on flagellin induced NLRC4 inflammasome activation in mouse bone marrow-derived macrophages(BMDMs).Methods:Mouse BMDMs were divided into control group,LPS group,LPS+flagellin group and LPS+acacetin+flagellin group.All groups were added with complete medium,then primed with LPS(50 ng/mL)for 3 hrs except the control group,whereafter,LPS+flagellin group was treated with flagellin(10μmol/L)for 0.5 hr and LPS+acacetin+flagellin group was treated with acacetin(10μmol/L)for 0.5 hr following by flagellin(10μmol/L)for 0.5 hr.Pro-caspase-1,pro-IL-1βin cell lysate and caspase-1,IL-1βin supernatant were detected by Western blot(WB).IL-1β,IL-18 and TNF-αin supernatant were measured by enzyme-linked immunosorbent assay(ELISA).And the activity of LDH in supernatant was assessed by LDH test kit.Results:Compared with the control group,in LPS+flagellin group,the expression of caspase-1,IL-1βprotein in supernatant were significantly increased(all P-values<0.05),but the differences of the expression of pro-caspase-1 and pro-IL-1βprotein in cell lysate were not significant.Compared with LPS+flagellin group,in LPS+acacetin+flagellin group,the expression of caspase-1,IL-1βprotein in supernatant were significantly reduced(all P-values<0.05),while the differences of the expression of pro-caspase-1 and pro-IL-1βprotein in cell lysate were not significant.ELISA showed that compared with the control group,the levels of IL-1β,IL-18,and TNF-αand the activity of LDH in supernatant of LPS+flagellin group were significantly increased(all P-values<0.05).Compared with LPS+flagellin group,in LPS+Acacetin+flagellin group,the level of IL-1βin supernatant was significantly decreased(P<0.05),meanwhile,the decreases of the levels IL-18,TNF-αand the activity of LDH were not significant.Conclusions:We found that Acacetin can effectively inhibit flagellin induced NLRC4 inflammasome activation and reduce cell damage in mouse BMDMs.
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