Inhibition of acacetin on AIM2 inflammasome activation in mouse BMDMs
Yeledan Mahan
WANG Zhaoxia
WU Xuanxia
BU Juan
Abstract：Objective To investigate the effects of acacetin on absent in melanoma 2(AIM2)inflammasome activa-tion co-stimulated by lipopolysaccharide(LPS)and poly(dA:dT)in mouse bone marrow-derived macrophages(BMD-Ms).Methods BMDMs from mouse femur were isolated and cultured,and then they were divided into the control group,LPS group,LPS + poly(dA:dT)group,and LPS + acacetin + poly(dA:dT)group,respectively.All groups were added with medium,then treated with 50 ng/mL LPS for 3 h except the control group,whereafter,LPS + poly(dA:dT)group was treated with poly(dA:dT)(10 μmol/L)for 0.5 h and LPS + acacetin + poly(dA:dT)group was treated with acacetin(10 μmol/L)for 0.5 h followed by stimulation with poly(dA:dT)(10 μmol/L)for 0.5 h.Western blotting was used to detect pro-Caspase-1 and pro-IL-1β protein expression in BMDMs lysate and Caspase-1 and IL-1β protein expres-sion in the supernatant.ELISA was applied to measure the levels of IL-18,IL-1β,and TNF-α,and the lactate dehydroge-nase(LDH)assay was used to measure cytotoxicity in the supernatant.Results No significant differences were found in the relative protein expression levels of pro-Caspase-1 or pro-IL-1β in the cell lysates among all groups(all P＞0.05).Compared with the control group and the LPS group,the relative protein expression levels of Caspase-1 and IL-1β in the su-pernatant of the LPS + poly(dA:dT)group and LPS + acacetin + poly(dA:dT)group significantly increased,with more significant changes in the LPS+poly(dA:dT)group(all P＜0.05).Compared with the control group and the LPS group,the protein expression levels of IL-1β and IL-18 in the supernatant of the LPS + poly(dA:dT)group and LPS + acacetin + poly(dA:dT)group increased,and the increase of IL-1β in the LPS+poly(dA:dT)group was more significant(all P＜0.05).Compared with the control group,the expression levels of TNF-α and the concentrations of LDH in the supernatant of the other three groups all increased,and the increase in the LPS + poly(dA:dT)group and LPS + acacetin + poly(dA:dT)group was more significant(all P＜0.05).Conclusion Acacetin exerts an inhibitory effect on the activation of the AIM2 inflammasome induced by LPS and poly(dA:dT)in mouse BMDMs,and it can reduce the protein expression of Cas-pase-1 and IL-1β in the cells,thereby alleviating the inflammatory response.
Keywords：acacetinabsent in melanoma 2inflammasomeinflammatory responsebone marrow-derived macro-phages
Publication Date：2023-12-15
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 30-33,38 )