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褪黑素对人腹膜间皮细胞上皮-间质转化的作用及机制OA北大核心CSTPCD

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

中文摘要英文摘要

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 DCNgene may be an important target to inhibit the EMT of these cells.

刘珊;张明霞;刘伦志

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

临床医学

褪黑素人腹膜间皮细胞核心蛋白聚糖上皮-间质转化

melatoninhuman peritoneal mesothelial celldecorinepithelial-mesenchymal transdifferentiation

《中国医科大学学报》 2024 (003)

235-239 / 5

湖北省教育厅科学研究计划(B2021155)

10.12007/j.issn.0258-4646.2024.03.008

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