大豆苷元减轻高糖诱导的肾小管上皮细胞焦亡  

Daidzein alleviates high glucose-induced pyroptosis of renal tubular epithelial cells

作　　者：陈才伟 吴月平[1] 王转锁[2] 罗强 邢博真 CHEN Caiwei;WU Yueping;WANG Zhuansuo;LUO Qiang;XING Bozhen(Department of Medical Healthcare,the First Affiliated Hospital of Hainan Medical College,Haikou 570102,China;Department of Endocrinology,the First Affiliated Hospital of Hainan Medical College,Haikou 570102,China)

机构地区：[1]海南医学院第一附属医院医疗保健科,海南海口570102 [2]海南医学院第一附属医院内分泌科,海南海口570102

出　　处：《基础医学与临床》2023年第12期1801-1807,共7页Basic and Clinical Medicine

基　　金：海南省医药卫生科研项目(2001320244A2014)。

摘　　要：目的探讨大豆苷元(daidxein,DAI)调节NOD样受体蛋白3(NOD-like receptor protein 3,NLRP3)/半胱天冬酶-1(caspase-1)信号通路对高糖诱导的肾小管上皮细胞焦亡的影响。方法将HK-2细胞分为:对照组(NC组)(5.5 mmol/L D-葡萄糖)、HG组(30 mmol/L D-葡萄糖)、DAI-L、DAI-M、DAI-H组(30 mmol/L D-葡萄糖分别和25、50、100μmol/L DAI孵育HK-2细胞)、DAI-H+LPS组(30 mmol/L D-葡萄糖、100μmol/L DAI和1μg/mL LPS共同孵育HK-2细胞)。MTT法检测HK-2细胞增殖;流式细胞术检测HK-2细胞凋亡;ELISA法检测HK-2细胞炎性因子白介素-1β(IL-1β)、白介素18(IL-18)水平;采用扫描电镜观察细胞焦亡形态;免疫荧光染色检测细胞焦亡相关蛋白;Western blot检测NLRP3、cleaved-caspase-1、GSDMD-N蛋白表达。结果NC组细胞呈圆球形,边界比较规则,而HG组细胞肿胀变大,边界不规则;HG组较NC组HK-2细胞吸光度A值(490 nm)显著下降(P<0.05),HK-2细胞凋亡率、IL-1β、IL-18含量、NLRP3、cleaved-caspase-1、GSDMD-N蛋白水平显著上调(P<0.05);DAI处理后细胞肿胀现象改善,HK-2细胞A值(490 nm)依次显著上升(P<0.05),HK-2细胞凋亡率、IL-1β、IL-18含量、NLRP3、cleaved-caspase-1、GSDMD-N蛋白水平依次显著降低(P<0.05),且DAI治疗效果呈现剂量依赖型,LPS消除了DAI-H对高糖诱导的肾小管上皮细胞焦亡的有利影响。结论DAI可缓解高糖诱导的肾小管上皮细胞焦亡。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 25,50,100μmol/L DAI,respectively),and DAI-H+LPS group(HK-2 cells were incubated with 30 mmol/L D-glucose,100μmol/L DAI and 1μg/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 observed by scanning electron microscope.Immunofluorescence staining was applied to detect pyroptosis related proteins.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-1β,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 induced apoptosis of renal tubular epithelial cells.Conclusions DAI may alleviate pyroptosis of renal tubular epithelial cells induced by high glucose through inhibition of NLRP3/caspase-1 signaling pathway.

关 键 词：大豆苷元 NLRP3/caspase-1信号通路 肾小管上皮细胞 细胞焦亡 

分 类 号：R285[医药卫生—中药学]