ABSTRACT
Objective To investigate the impact of daidzein(DAI)on the pyroptosis of renal tubular epithelial cells induced by high glucose by regulating NOD-like receptor protein 3(NLRP3)/caspase-1 signal pathway.Methods HK-2 cells were divided into control group(NC group)(5.5 mmol/L D-glucose),HG group(30 mmol/L D-glucose),DAI-L,DAI-M,DAI-H groups(HK-2 cells were incubated with 30 mmol/L D-glucose and 25,50,100 μmol/L DAI,respectively),and DAI-H+LPS group(HK-2 cells were incubated with 30 mmol/L D-glucose,100 μmol/L DAI and 1 μg/mL LPS).MTT assay was applied to detect the cytotoxicity and proliferation of HK-2 cells;the apoptosis of HK-2 cells was detected by flow cytometry.The level of interleukin-1β(IL-1β)and inter-leukin-18(IL-18)in HK-2 cells was detected by ELISA.The morphology of pyroptosis cells was ob-served by scanning electron microscope.Immunofluorescence staining was applied to detect pyroptosis related pro-teins.The expression of NLRP3,cleaved casase-1 and GSDMD-N was detected by Western blot.Results In NC group,the cells were spherical with regular boundaries,while in HG group,the cells swelled and became larger with irregular boundaries;the OD value(490 nm)of HK-2 cells in HG group was obviously lower than that in NC group(P＜0.05),the apoptosis rate,IL-1β,IL-18 contents,NLRP3,cleaved casase-1,GSDMD-N protein level of HK-2 cells were obviously higher(P＜0.05);After DAI treatment,the swelling of cells was alleviated,the A value(490 nm)of HK-2 cells increased significantly(P＜0.05),the apoptosis rate of HK-2 cells,IL-1 β,IL-18 content,NLRP3,cleaved-caspase-1 and GSDMD-N protein levels were significantly decreased(P＜0.05)and the therapeutic effect of DAI was dose-dependent;LPS eliminated the beneficial effect of DAI-H on high glucose in-duced apoptosis of renal tubular epithelial cells.Conclusions DAI may alleviate pyroptosis of renal tubular epithe-lial cells induced by high glucose through inhibition of NLRP3/caspase-1 signaling pathway.