Role Of KCNQ1 In The Process Of MSU Crystals Activating Human Monocyte-macrophage To Release IL-1β

Objective: To explore the possible role of KCNQ1 in the process of monosodium urate monohydrate(MSU)crystals activating human monocyte-macrophage to release IL-1β.Methods: THP-1 were treated with 60μg/ml PMA for 72 h,thenwe got the human monocyte-macrophage.Detect the expression of KCNE1 in human monocyte-macrophage.Divide them into four groups: normal control groups,MSU crystals groups,MSU crystals plus KCNQ1 channel activators(MSU+MLL-277)groups,MSU crystals plus channel inhibitors(MSU+Chromanol293B)groups.The concentrations of activators were 20 m M,and inhibitors were 100 m M.Quality of MSU crystals in each group was 100μg/ml.Pretreat the cells with corresponding regents(activators or inhibitors)for 30 mins,then add MSU crystals into each group.6 hours later,detect the concentration of IL-1beta in supernatant in each group by means of ELISA,and the expression of pro-IL-1beta,pro-IL-18,TNF-α in each group by means of fluorescence quantitive real-time PCR.2 hours later,discard the supernatant,the cells were lysed by 10% nitricacid.Then,test the concentrations of K+in each group by atomic absorbancespectroscopy.We also tested the concentrations of intracellular ATP by ATP assay kit,after stimulating for 2h.Results: The expression level of KCNE1 was higher than KCNQ1 in human monocyte-macrophage.After stimulating by MSU crystals for 6 hours,the concentrations of IL-1β in group(MSU+ML-277)were much higher than group(MSU)(p<0.01),and the concentrations in group(MSU+293B)had no significant statistic differences with group(MSU).The expressions of pro-IL-1beta,pro-IL-18,TNF-α in different groups had no significant difference either.After stimulated by MSU crystals for 2 hours,the concentrations of intracellular K+ in group(MSU+ML-277)were lower than group(MSU)(p<0.01).And concentrations in group(MSU+293B)seemed to be higher than group(MSU).Although we didn’t observe the statistically significant difference under the existing condition,we believed that if the sample size was enlarged,the difference would be seen.The concentrations of intracellular ATP in control group were much higher than in MSU crystals treatment group.Conclusion:1.Under normal circumstances,KCNQ1 may play limited role in the release of IL-1βfrom monocyte-macrophage;but when it isexcessively activated,the release of IL-1β would increase significantly.2.The abnormal regulating effect of KCNE1 on KCNQ1 in producing mature IL-1βis worthy studying.