白藜芦醇干预白蛋白诱导人近端肾小管上皮细胞HK-2细胞焦亡相关蛋白的表达  

Effects of resveratrol on albumin-induced pyroptosis-related proteins in HK-2 cells

作　　者：李方 曹建民[2] 翟鹏飞[1] 王建设[1] 洪达[1] Li Fang;Cao Jianmin;Zhai Pengfei;Wang Jianshe;Hong Da(Physical Education College,Henan Normal Unversity,Xinxiang 453007,Henan Province,China;School of Sport Science,Beijing Sport University,Beijing 100084,China)

机构地区：[1]河南师范大学体育学院,河南省新乡市453007 [2]北京体育大学运动人体科学学院,北京市100084

出　　处：《中国组织工程研究》2023年第14期2164-2169,共6页Chinese Journal of Tissue Engineering Research

基　　金：河南省重点研发与推广专项(科技攻关)项目(212102310261),项目负责人:李方。

摘　　要：背景:白藜芦醇有肾脏保护作用,但其对蛋白尿负荷的肾小管上皮细胞炎性损伤的保护机制尚不明确。目的:探究白藜芦醇对白蛋白诱导的人近端管上皮细胞HK-2细胞焦亡相关蛋白表达的影响。方法:采用白蛋白诱导HK-2细胞炎性损伤,分别使用白藜芦醇、SIRT1抑制剂EX527进行预处理,MTT法检测细胞活力,Western blot检测SIRT1、NLRP3、ASC、caspase-1的蛋白表达,免疫荧光双染观察NLRP3和ASC在肾小管的共定位,Hoechst33342/PI染色法观察细胞焦亡情况,ELISA检测细胞上清中炎症因子白细胞介素1β和白细胞介素18水平。结果与结论:(1)白蛋白减低HK-2细胞活力,显著升高白细胞介素1β、白细胞介素18水平和NLRP3炎性小体蛋白表达(P <0.01),增加细胞焦亡;(2)白藜芦醇可显著提高HK-2细胞中SIRT1蛋白表达(P <0.01),显著降低NLRP3、ASC和caspase-1蛋白表达(P <0.05)及白细胞介素1β、白细胞介素18水平(P <0.01),减少细胞焦亡;(3)SIRT1抑制剂可显著提高NLRP3炎性小体的表达,增加细胞焦亡;(4)结果表明,白藜芦醇调节HK-2细胞焦亡相关蛋白改善白蛋白诱导的肾小管上皮细胞炎性损伤。BACKGROUND:Resveratrol exerts protective effects against renal diseases,while its protective mechanism against inflammatory injury of renal tubular epithelial cells induced by albumin is still unclear.OBJECTIVE:To explore the effect of resve ratrol on pyroptosis-related protein expression in HK-2 cells.METHODS:Albumin was used to induce inflammatory damage in HK-2 cells.Resve ratrol and sirtuins(SIRT1) inhibitor EX527 were used as pretreatments.MTT assay was used to assess cell viability.Western blot was applied to test the protein expression of SIRT1,NOD-like receptor protein 3(NLRP3),caspase-1 and apoptosis-associated speck-like protein(ASC).Immunofluorescence double staining was used to observe the co-localization of NLRP3 and ASC.Hoechst33342/PI staining was used to observe pyrolysis.Levels of interleukin-1β and interleukin-18 were tested by using ELISA.RESULTS AND CONCLUSION:Albumin co uld significantly decrease cell viability,elevate the expressions of interleukin-1β,interleukin-18 and NLRP3 inflammasome(P<0.01),and increase pyroptosis in HK-2 cells.However,resveratrol significantly upregulated the protein expression of SIRT1 in HK-2 cells(P<0.01),dow n regulated the expression of NLRP3,ASC and caspase-1(P <0.05) and the levels of inte rleukin-1β and interleukin-18(P<0.01),and attenuated albumin-induced pyroptosis.SIRT1 inhibitor could remarkably increase the expression of NLRP3 inflammasome and increase pyrolysis.To conclude,resve ratrol modulates HK-2 cell pyroptosis-related proteins to ameliorate albumin-induced inflammatory injury in renal tubular epithelial cells.

关 键 词：白藜芦醇 肾小管上皮细胞 NLRP3炎性小体 细胞焦亡 白蛋白 

分 类 号：R459.9[医药卫生—治疗学] R334.1[医药卫生—临床医学] R363.1