Effect of deubiquitinase ABRO1 on IL-1β release of monocyte macrophage induced by listeriolysin O
Objective To observe the effect of deubiquitinase Abraxas brother 1(ABRO1)on the release of interleu-kin(IL)-1β from monocyte macrophage J774A.1 in mice induced by listeriolysin O(LLO),and to explore the relevant mechanisms.Methods J774A.1 cells were cultured and infected with wild-type Listeria monocytogenes(LM)strains with LLO(wild-type group),hly gene deficient(Δhly)LM strains with LLO knockout(gene deficient group),and Δhly strains supplemented with hly gene LM strains(replenishment group),respectively.Western blotting was used to detect ABRO1 protein,and ELISA was used to detect IL-1β in the supernatant of cell culture medium.We divided J774A.1 cells into NI group(uninfected LM strain),WT group(infected WT LM strain),and Δhly group(infected Δhly LM strain),and cells in each group were transfected with NCsiRNA and ABRO1siRNA,respectively.The IL-1β in the super-natant of the cell culture medium was detected by ELISA,and Western blotting was used to detect inflammasome-related molecules Caspase-1,p20,and IL-1β,and p17.Results Over the infection time,the expression of ABRO1 and IL-1β levels gradually increased in the wild-type group and the replenishment group,reaching their highest levels at 120 minutes of infection,and both were higher than those in the gene deficient group(all P<0.05).There were no significant changes in ABRO1 expression or IL-1β levels at different time points in the gene deficient group.The level of IL-1β and expression of p20 and p17 in the supernatant of cells transfected with ABRO1 siRNA in the WT group were lower than those in cells transfected with NC siRNA(all P<0.05).The level of IL-1β and expression of p20 and p17 in the supernatant of cells transfected with NC siRNA in the WT group were higher than those in the NI group(all P<0.05).The level of IL-1β and expression of p20 and p17 in the supernatant of cells transfected with NC siRNA in the Δhly group were lower than those in the WT group(all P<0.05).Conclusions Under the induction of LLO,the expression of ABRO1 increased in J774A.1 cells.After down-regulating ABRO1 expression,the release of IL-1β in LLO-induced J774A.1 cells decreased;LLO may promote LLO-mediated IL-1β release by activating inflammasomes.
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