| Abstrakt: |
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 (10 μmol/L) for 0.5 hr and LPS + acacetin + flagellin group was treated with acacetin (10 μmol/L) for 0.5 hr following by flagellin (10 μmol/L) for 0.5 hr. Pro-caspase-1, pro-IL-1β in cell lysate and caspase-1, IL-1β in supernatant were detected by Western blot (WB). IL-1β, IL-18 and TNF-α in 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-1β protein in supernatant were significantly increased (all P-values<0.05), but the differences of the expression of pro-caspase-1 and pro-IL-1β protein in cell lysate were not significant. Compared with LPS + flagellin group, in LPS + acacetin + flagellin group, the expression of caspase-1, IL-1β protein in supernatant were significantly reduced (all P-values<0.05), while the differences of the expression of pro-caspase-1 and pro-IL-1β protein in cell lysate were not significant. ELISA showed that compared with the control group, the levels of IL-1β, IL-18, and TNF-α and 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-1β in supernatant was significantly decreased (P<0.05), meanwhile, the decreases of the levels IL-18, TNF-α and 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. [ABSTRACT FROM AUTHOR] |
