表没食子儿茶素没食子酸酯抑制人腹膜间皮细胞上皮-间质转化的作用及机制  

Effect and Mechanism of Epigallocatechin Gallate on Inhibiting Epithelial-Mesenchymal Transition of Human Peritoneal Mesothelial Cells

作　　者：谢斌 洪慧 帅欢[1] 张林[2] XIE Bin;HONG Hui;SHUAI Huan;ZHANG Lin(The First Affiliated Hospital of Hunan Traditional Chinese Medical College,Zhuzhou,Hunan,China 412000;The Fourth Hospital of Changsha,Changsha,Hunan,China 410006)

机构地区：[1]湖南中医药高等专科学校附属第一医院,湖南株洲412000 [2]湖南省长沙市第四医院,湖南长沙410006

出　　处：《中国药业》2023年第5期65-71,共7页China Pharmaceuticals

基　　金：湖南省卫生计生委科研计划课题项目[B20180473]。

摘　　要：目的探讨表没食子儿茶素没食子酸酯(EGCG)对人腹膜间皮细胞(HPMCs)上皮-间质转化(EMT)的作用及机制。方法收集新开管的腹膜透析和腹膜透析1年以上患者的HPMCs,分别采用逆转录实时荧光定量聚合酶链反应(qRT-PCR)法和免疫印迹(Western blot)法检测紧密连接蛋白1(ZO-1)、E-钙黏蛋白(E-cadherin)、铁死亡抑制蛋白1(FSP1)mRNA和蛋白表达水平。采用HPMCs细胞系HMrSV5进行体外实验研究,分别加入20,40,60,80,100μg/mL EGCG,检测EGCG对HMrSV5的细胞毒性。通过200μg/mL和500μg/mL晚期糖基化终产物(AGEs)构建HPMCs EMT模型,将HMrSV5细胞分为对照组(AGEs造模前无处理)、实验组(AGEs造模前2 h加入EGCG共孵育)、si-NC+EGCG组(转染si-NC后,AGEs造模前2 h加入EGCG共孵育)、si-Notch3+EGCG组(转染si-Notch3后,AGEs造模前2 h加入EGCG共孵育),检测ZO-1,E-cadherin,FSP1,Notch3 mRNA和蛋白表达水平。结果与新开管的腹膜透析患者比较,腹膜透析1年以上患者HPMCs的ZO-1及E-cadherin mRNA和蛋白表达水平均显著升高,FSP1及Notch3 mRNA表达水平均显著降低(P<0.05)。与正常HMrSV5比较,加入500μg/mL AGEs诱导HMrSV5后,ZO-1及E-cadherin mRNA和蛋白表达水平均显著升高,FSP1 mRNA和蛋白表达水平均显著降低(P<0.05)。与对照组比较,实验组ZO-1及E-cadherin mRNA和蛋白表达水平均显著降低,FSP1及Notch3 mRNA和蛋白表达水平均显著升高(P<0.05)。与si-NC+EGCG组比较,si-Notch3+EGCG组Notch3及FSP1 mRNA和蛋白表达水平均显著降低,ZO-1及E-cadherin mRNA和蛋白表达水平均显著升高(P<0.05)。腹膜透析患者HPMCs中Notch3表达水平与ZO-1和E-cadherin呈负相关(r=-0.8477,-0.3822,P<0.05),与FSP1呈正相关(r=0.6626,P<0.05)。结论EGCG通过Notch3抑制AGEs诱导的HPMCs EMT,发挥对腹膜透析患者的保护作用。Objective To investigate the effect and mechanism of epigallocatechin gallate(EGCG)on inhibiting the epithelial-mesenchymal transiton(EMT)of human peritoneal mesothelial cells(HPMCs).Methods HPMCs from patients starting peritoneal dialysis(PD)and underwent PD for more than one year were collected,and the expression levels of zonula occluden-1(ZO-1),E-cadherin and ferroptosis suppressor protein 1(FSP1)mRNA and protein were detected by the reverse transcription real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)and Western blot,respectively.HPMCs cell line HMrSV5 was used for in vitro study,and 20,40,60,80,100μg/mL EGCG were added into HMrSV5 cell culture fluid respectively to detect the cytotoxicity of EGCG to HMrSV5 cells.The EMT model of HPMCs was constructed by 200μg/mL and 500μg/mL advanced glycation end products(AGEs).HMrSV5 cells were divided into the control group(no treatment before modeling by AGEs),the test group(EGCG was added to co-incubate with HMrSV5 cells 2 h before modeling by AGEs),the si-NC+EGCG group(EGCG was added to co-incubate with HMrSV5 cells after transfection of si-NC and 2 h before modeling by AGEs)and the si-Notch3+EGCG group(EGCG was added to co-incubate with HMrSV5 cells after transfection of si-Notch3 and 2 h before modeling by AGEs).The expression levels of ZO-1,E-cadherin,FSP1,Notch3 mRNA and protein were detected.Results Compared with those in the patients starting PD,the expression levels of ZO-1 and E-cadherin mRNA and protein of HPMCs in patients underwent PD for more than one year were significantly higher,and the expression levels of FSP1 and Notch3mRNA of HPMCs in patients underwent PD for more than one year were significantly lower(P<0.05).Compared with those in the normal HMrSV5 cells,the expression levels of ZO-1 and E-cadherin mRNA and protein in the HMrSV5 cells induced by500μg/mL AGEs were significantly higher,and the expression levels of FSP1 mRNA and protein in the HMrSV5 cells induced by 500μg/mL AGEs were significantly lower(P<0.05).Compared

关 键 词：表没食子儿茶素没食子酸酯 人腹膜间皮细胞 上皮-间质转化 晚期糖基化终产物 腹膜透析 

分 类 号：R965[医药卫生—药理学] R979.4[医药卫生—药学] R692