TXNIP/NLRP3通路在妊娠期糖尿病胎盘滋养层细胞迁移侵袭中的作用研究  

The role of TXNIP/NLRP3 pathway in migration and invasion of placental trophoblasts in gestational diabetes mellitus

作　　者：时艳梅[1] 陈晓芳[1] 张素芹 Shi Yanmei;Chen Xiaofang;Zhang Suqin(Department of Gynecology and Obstetrics,Yantai Yantaishan Hospital,Yantai 264003,China)

机构地区：[1]烟台市烟台山医院产科,264003

出　　处：《中华内分泌外科杂志》2020年第5期416-421,共6页Chinese Journal of Endocrine Surgery

基　　金：烟台市科技计划项目(2020MSGY088)。

摘　　要：目的探讨硫氧还蛋白结合蛋白(thioredoxin-interacting protein,TXNIP)/核苷酸结合寡聚化结构域样受体蛋白3(NLR family pyrin domain containing 3,NLRP3)通路在妊娠期糖尿病(gestational diabetes mellitus,GDM)患者胎盘组织中的表达及其在胎盘滋养层细胞迁移和侵袭中的作用。方法收集25例GDM孕妇(GDM组)和25例糖耐量正常孕妇(正常组)的胎盘组织,将体外培养的人绒毛膜滋养层细胞分为对照组(未转染)、siNC组(转染阴性对照)、siTXNIP组(转染siRNA-TXNIP)、siTXNIP+pcDNA组(共转染siRNA-TXNIP和pcDNA3.1空载体质粒)和siTXNIP+NLRP3组(共转染siRNA-TXNIP和pcDNA3.1-Flag-NLRP3过表达质粒),采用实时荧光定量PCR检测TXNIP和NLRP3 mRNA表达水平,免疫印迹法检测TXNIP和NLRP3蛋白表达水平,噻唑蓝法检测细胞活力,划痕实验检测细胞迁移能力,Transwell小室实验检测细胞侵袭。结果与正常组比较,GDM组患者胎盘组织中TXNIP和NLRP3在mRNA和蛋白水平上的表达均明显升高(P<0.05);与对照组或siNC组比较,siTXNIP组细胞中TXNIP在mRNA和蛋白水平上的表达明显降低,而细胞活力和迁移率均明显升高,且穿膜细胞数明显增多(P<0.05);而siNC组和对照组比较,上述各指标差异均无统计学意义(P>0.05);与siTXNIP组或siTXNIP+pcDNA组比较,siTXNIP+NLRP3组细胞中NLRP3在mRNA和蛋白水平上的表达明显升高,而细胞活力和迁移率均明显降低,且穿膜细胞数明显减少(P<0.05);但siTXNIP+pcDNA组和siTXNIP组之间差异无统计学意义(P>0.05)。结论TXNIP/NLRP3通路与GDM发生发展密切相关,TXNIP可通过激活NLRP3抑制胎盘滋养层细胞迁移侵袭。Objective To investigate the expressions of thioredoxin-interacting protein(TXNIP)/NLR family pyrin domain containing 3(NLRP3)pathway in placental tissues of gestational diabetes mellitus(GDM)patients and its role in the migration and invasion of placental trophoblast cells.Methods The placenta tissues of 25 GDM pregnant women(GDM group)and 25 normal glucose tolerance pregnant women(normal group)were collected.The cultured human chorionic trophoblast cells were divided into control group(untransfected),siNC group(negative control),siTXNIP group(siRNA-TXNIP),siTXNIP+pcDNA group(co transfected with siRNA-TXNIP and pcDNA3.1 empty vector plasmids)and siTXNIP+NLRP3 group(co transfected with siRNA-TXNIP and pcDNA3.1-Flag-NLRP3 over expression plasmids).The expressions of TXNIP and NLRP3 mRNAs were detected by real-time fluorescence quantitative PCR,and the expressions of TXNIP and NLRP3 proteins were detected by Western blot.Cell viability was measured by thiazole blue method.Cell migration ability was detected by scratch test,and cell invasion was detected by Transwell’s cell experiment.Results Compared with those in the normal group,mRNA and protein expression levels of TXNIP and NLRP3 in the placenta of patients in GDM group were significantly higher(P<0.05).Compared with those in the control group or siNC group,mRNA and protein expression levels of TXNIP in siTXNIP group were significantly lower,however,the cell viability and migration rate were significantly higher,and the number of transmembrane cells was significantly higher(P<0.05);but there was no significant difference in the above indexes between siNC group and the control group(P>0.05).Compared with those in siTXNIP group or siTXNIP+pcDNA group,mRNA and protein expression levels of NLRP3 in siTXNIP+NLRP3 group were significantly higher,however,the cell viability and migration rate were significantly lower,and the number of transmembrane cells was significantly lower(P<0.05);but there was no significant difference in the above indexes between siTXNIP+pcDNA

关 键 词：妊娠期糖尿病 硫氧还蛋白结合蛋白 核苷酸结合寡聚化结构域样受体蛋白3 胎盘滋养层细胞 迁移 侵袭 

分 类 号：R71[医药卫生—妇产科学] R587.1[医药卫生—临床医学]