褪黑素对人腹膜间皮细胞上皮-间质转化的作用及机制  

Effect and mechanism of melatonin on the epithelial-mesenchymal transition of human peritoneal mesenchymal cells

作　　者：刘珊 张明霞 刘伦志 LIU Shan;ZHANG Mingxia;LIU Lunzhi(Department of Nephrology,Minda Hospital of Hubei Minzu University,Hubei Provincial Clinical Medical Research Center for Nephropathy,Enshi 445000,China)

机构地区：[1]湖北民族大学附属民大医院肾内科,湖北省肾脏病临床医学研究中心,湖北恩施445000

出　　处：《中国医科大学学报》2024年第3期235-239,共5页Journal of China Medical University

基　　金：湖北省教育厅科学研究计划(B2021155)。

摘　　要：目的探讨褪黑素对人腹膜间皮细胞上皮-间质转化(EMT)的作用及机制。方法培养人腹膜间皮细胞株HMrSV5,构建核心蛋白聚糖(DCN)过表达和敲减质粒。将细胞分为正常对照组、TGF-β1组、TGF-β1+褪黑素组、TGF-β1+褪黑素+siDCN组、TGF-β1+褪黑素+siNC组、TGF-β1+DCN组。采用CCK-8法检测细胞增殖存活率;采用蛋白免疫印迹和实时定量PCR检测DCN、TGF-β1、Smad2、E-cadherin蛋白和mRNA表达水平。结果CCK-8结果显示,TGF-β1+褪黑素组、TGF-β1+褪黑素+siDCN组、TGF-β1+DCN组细胞存活率均较TGF-β1组高,TGF-β1+褪黑素组较TGF-β1+褪黑素+siDCN组细胞存活率更高(均P<0.05)。蛋白免疫印迹及实时定量PCR表明,TGF-β1组较对照组DCN和E-cadherin表达明显下调,TGF-β1+褪黑素组与TGF-β1+褪黑素+siDCN组较TGF-β1组DCN、E-cadherin蛋白及mRNA表达上调,TGF-β1、Smad2蛋白及mRNA表达下调;TGF-β1+褪黑素组较TGF-β1+褪黑素+siDCN组进一步上调E-cadherin、DCN蛋白及mRNA表达,进一步下调TGF-β1、Smad2蛋白及mRNA表达(均P<0.05)。结论褪黑素可延缓人腹膜间皮细胞EMT进程,DCN基因可能是抑制该细胞发生EMT的一个重要靶点。Objective To investigate the effect and mechanism of melatonin on the epithelial-mesenchymal transition(EMT)of human peritoneal mesothelial cells.Methods A human peritoneal mesoepithelial cell line(HMrSV5)was cultured.Decorin(DCN)overex-pression and knockout plasmids were constructed.The cells were divided into the normal,TGF-β1,TGF-β1+melatonin,TGF-β1+mela-tonin+siDCN,TGF-β1+melatonin+siNC,and TGF-β1+DCN groups.The cell proliferation and survival rates were determined using Cell Counting Kit-8(CCK-8).The protein and mRNA expression levels of DCN,TGF-β1,Smad2,and E-cadherin were detected by Western blotting and real-time polymerase chain reaction(PCR),respectively.Results The CCK-8 assay showed that the cell survival rates were higher in the TGF-β1+melatonin,TGF-β1+melatonin+siDCN,and TGF-β1+DCN groups than the TGF-β1 group(P<0.05).The cell sur-vival rate was higher for the TGF-β1+melatonin group than the TGF-β1+melatonin+siDCN group(P<0.05).Western blotting and real-time PCR showed DCN and that E-cadherin were significantly down-regulated in the TGF-β1 group compared with the control group(P<0.05).Compared with the TGF-β1 group,the TGF-β1+melatonin and TGF-β1+melatonin+siDCN groups showed up-regulated E-cadherin and DCN expression levels and down-regulated TGF-β1 and Smad2 expression levels(P<0.05).Compared with the TGF-β1+melato-nin+siDCN group,the TGF-β1+melatonin+siDCN group showed up-regulated E-cadherin and DCN expression levels and down-regulated TGF-β1 and Smad2 expression levels(P<0.05).Conclusion Melatonin can delay the EMT of human peritoneal mesoepithelial cells,and the DCN gene may be an important target to inhibit the EMT of these cells.

关 键 词：褪黑素 人腹膜间皮细胞 核心蛋白聚糖 上皮-间质转化 

分 类 号：R656.4[医药卫生—外科学]