Objective This paper is to investigate the effect of adenosine A3 receptor(A3AR)agonist 2-Cl-IB-MECA on human intestinal epithelial cell pyroptosis and its molecular mechanism.Methods HT-29 cells were divided into the control group(no treatment),the lipopolysaccharide(LPS)group(LPS at a final concentration of 10 μg/mL),the 30 nmol/L MECA group(2-Cl-IB-MECA at a final concentration of 30 nmol/L on the basis of the LPS group),and the 100 nmol/L MECA group(2-Cl-IB-MECA at a final concentration of 100 nmol/L on the basis of the LPS group).Lactate dehydrogenase(LDH)activity assay and propidium iodide(PI)fluorescence staining were used to detect cell pyroptosis.The expression of IL-1β in cell culture supernatant was detected by ELISA.The expression of phosphorylated NF-κB p65(p-NF-κB p65),NOD-like receptor thermal protein domain associated protein 3(NLRP3),caspase-1,gasdermin D(GSDMD),and IL-1β were detected by Western blotting.Results Compared with the control group,the number of PI-stained positive cells and LDH activity are increased,and the expression of p-NF-κB p65,NLRP3,caspase-1,GSDMD,and IL-1β are elevated in the LPS group,with statistically significant differences(P＜0.05).Compared with the LPS group,the number of PI-stained positive cells and LDH activity are reduced,and the expression of p-NF-κB p65,NLRP3,caspase-1,GSDMD,and IL-1β are decreased in the 2-Cl-IB-MECA group,with statistically significant differences(P＜0.05).Conclusion A3AR activation attenuates intestinal epithelial cell pyroptosis,whilch may be related to the inhibition of the NF-κB/NLRP3 signaling pathway.
Adenosine A3 receptorNOD-like receptor thermal protein domain associated protein 3Intestinal epithelial cellsCell pyroptosis
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