硫氧还蛋白1/硫氧还蛋白相互作用蛋白对腹膜间皮细胞核苷酸结合寡聚化结构域样受体蛋白3炎症小体的调控及机制研究  

The regulation and mechanism of thioredoxin 1 and thioredoxin interacting protein on nucleotide-binding oligomerized domain-like receptor protein 3 inflammasome in peritoneal mesothelial cells

作　　者：程锐 朱长才 Cheng Rui;Zhu Changcai(Department of Urology,Huangshi Aikang Hospital,Huangshi 435000,China;Department of Labor Health and Environmental Health,School of Public Health,Wuhan University of Science and Technology,Wuhan 430062,China)

机构地区：[1]黄石市爱康医院泌尿外科,黄石435000 [2]武汉科技大学医学院公共卫生学院劳动卫生与环境卫生学教研室,武汉430062

出　　处：《中国医师进修杂志》2024年第6期513-517,共5页Chinese Journal of Postgraduates of Medicine

摘　　要：目的探究硫氧还蛋白1(Trx1)/硫氧还蛋白相互作用蛋白(Txnip)对腹膜间皮细胞核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体的调控作用及可能机制。方法前瞻性采用随机数字表法将人腹膜间皮细胞(HMrSV5)分为三组,一组采用10%胎牛血清(FBS)培养液(空白组)、一组采用转染小干扰RNA(siRNA)阴性对照的4.25%腹膜透析液(PDS)与10%FBS培养液(si-NC组),一组采用转染靶向沉默Txnip siRNA的4.25%PDS与10%FBS培养液(si-Txnip组)。采用实时定量聚合酶链反应(qRT-PCR)、免疫印迹法(WB)测定各组Trx1、Txnip、NLRP3 mRNA与蛋白表达水平;采用MTT法测定各组培养不同时间点细胞增殖情况;采用酶联免疫吸附法(ELISA)测定上清液白细胞介素(IL)-1β、IL-6水平和半胱氨酸蛋白酶1(caspase-1)活性。结果si-NC组、si-Txnip组培养1、2、3 d HMrSV5增殖活性均低于空白组(P<0.05);并且si-Txnip组培养1、2、3 d HMrSV5增殖活性均高于si-NC组(0.402±0.009比0.372±0.010、0.554±0.016比0.482±0.008、0.700±0.013比0.590±0.010),差异有统计学意义(P<0.05)。si-NC组、si-Txnip组Txnip、NLRP3 mRNA表达均高于空白组(P<0.05);并且si-Txnip组Txnip、NLRP3 mRNA表达低于si-NC组(1.43±0.32比2.80±0.43、2.38±0.35比3.42±0.40),si-Txnip组Trx1 mRNA表达低于空白组、高于si-NC组(2.24±0.35比3.50±0.38、1.18±0.23),差异有统计学意义(P<0.05)。si-NC组、si-Txnip组Txnip、NLRP3蛋白表达均高于空白组(P<0.05);si-Txnip组Txnip、NLRP3蛋白表达低于si-NC组(0.453±0.108比0.754±0.116),si-Txnip组Trx1蛋白表达低于空白组、高于si-NC组(0.514±0.112比0.753±0.125、0.297±0.010),差异有统计学意义(P<0.05)。si-NC组、si-Txnip组上清液IL-1β、IL-6水平均高于空白组[(0.45±0.07)、(0.35±0.06)μg/L比(0.23±0.05)μg/L,(3.00±0.38)、(2.32±0.30)μg/L比(1.95±0.34)μg/L],si-Txnip组上清液IL-1β、IL-6水平低于si-NC组,差异有统计学意义(P<0.05)。si-Txnip组上清液caspase-1活性小于si-Objective To explore the regulatory effect and possible mechanism of thioredoxin 1(Trx1)/thioredoxin interacting protein(Txnip)on nucleotide-binding oligomerized domain-like receptor protein 3(NLRP3)inflammasome in peritoneal mesothelial cells.Methods Human peritoneal mesothelial cell line(HMrSV5)was divided into three groups by random number table method.One group was cultured with 10%fetal bovine serum(FBS)medium(the blank group),another group was cultured with 4.25%peritoneal dialysate(PDS)and 10%FBS medium with transfected small interfering RNA(siRNA)negative control(si-NC group),the last group was transfected with 4.25%PDS and 10%FBS culture medium(si-Txnip group).Quantitative real-time polymerase chain reaction(qRT-PCR)and Western blotting(WB)were used to determine the mRNA and protein expressions of Trx1,Txnip and NLRP3 in each group,respectively;MTT method was used to determine cell proliferation in each group at different time points.The levels of interleukin(IL)-1β,IL-6 and the activity of caspase-1 in the supernatant were determined by enzyme-linked immunosorbent assay(ELISA).Results The proliferative activity of HMrSV5 cultured for 1,2 and 3 d in si-NC group and si-Txnip group were lower than those in the blank group(P<0.05);the proliferative activity of HMrSV5 cultured for 1,2 and 3 d in si-Txnip group were higher than those in the si-NC group:0.402±0.009 vs.0.372±0.010,0.554±0.016 vs.0.482±0.008,0.700±0.013 vs.0.590±0.010,there were statistical differences(P<0.05).The mRNA expressions of Txnip and NLRP3 in si-NC group and si-Txnip group were higher than those in the blank group(P<0.05),and the mRNA expressions of Trx1 and NLRP3 in si-Txnip group were lower than those in the si-NC group:1.43±0.32 vs.2.80±0.43,2.38±0.35 vs.3.42±0.40;the mRNA expression of Trx1 in si-Txnip group were lower than that in the blank group and higher than that in the si-NC group:2.24±0.35 vs.3.50±0.38 and 1.18±0.23,there were statistical differences(P<0.05).The Txnip and NLRP3 protein expressions in the si-NC

关 键 词：腹膜间皮细胞 硫氧还蛋白1 硫氧还蛋白相互作用蛋白 核苷酸结合寡聚化结构域样受体蛋白3 

分 类 号：R363[医药卫生—病理学]