茶多酚EGCG通过调控miR-16-5p/含铜胺氧化酶1轴发挥对过氧化氢诱导的人心肌细胞凋亡的保护作用  

Protective Effect of EGCG on Hydrogen Peroxide-induced Apoptosis of Human Cardiomyocyte via Regulating miR-16-5p/AOC1 Axis

作　　者：毕樱馨 刘咸筠 孟祥龙 柳越 王梦媛 张妍 李占东[2] 殷玉和[1] 李皓[2] BI Yingxin;LIU Xianjun;MENG Xianglong;LIU Yue;WANG Mengyuan;ZHANG Yan;LI Zhandong;YIN Yuhe;LI Hao(School of Chemistry and Life Sciences,Changchun University of Technology,Changchun 130012,China;College of Food Engineering,Jilin Engineering Normal University,Changchun 130052,China;Department of Burns Surgery,The First Hospital of Jilin University,Changchun 130021,China)

机构地区：[1]长春工业大学化学与生命科学学院,吉林长春130012 [2]吉林工程技术师范学院食品工程学院,吉林长春130052 [3]吉林大学第一医院烧伤及皮肤创面修复外科,吉林长春130021

出　　处：《食品工业科技》2022年第7期376-383,共8页Science and Technology of Food Industry

基　　金：吉林工程技术师范学院博士启动项目(BSKJ201859,BSKJ201923)。

摘　　要：目的:探究茶多酚表没食子儿茶素没食子酸酯(EGCG)对过氧化氢(H_(2)O_(2))诱导人心肌细胞HCM凋亡的保护作用中的分子机制。方法:利用H_(2)O_(2)建立HCM细胞凋亡模型;通过caspase-3活性检测实验评价细胞凋亡水平;利用CCK-8法检测药物的细胞毒性;qPCR和Western blot用于检测含铜胺氧化酶1(AOC1)和miR-16-5p的表达水平;通过细胞转染调控AOC1和miR-16-5p在HCM细胞中的表达;通过TargetScan预测以及双荧光素酶报告基因实验验证miR-16-5p和AOC1 mRNA 3'UTR区的结合关系以及结合位点。结果:EGCG处理后可以显著抑制H_(2)O_(2)诱导的HCM细胞凋亡(P<0.05);在凋亡细胞中,AOC1表达显著下降,EGCG可以显著上调AOC1的表达,而敲减AOC1显著逆转EGCG对凋亡的抑制(P<0.05);同时,在凋亡细胞中,miR-16-5p表达显著上升,EGCG可以显著抑制miR-16-5p的表达,而上调miR-16-5p显著逆转了EGCG对凋亡的抑制(P<0.05);上调miR-16-5p显著逆转了EGCG对凋亡细胞中AOC1表达的促进;最后,miR-16-5p可以靶向抑制AOC1的表达,而过表达AOC1显著逆转了miR-16-5p诱导的HCM细胞凋亡。结论:EGCG通过调控miR-16-5p/AOC1轴发挥了对H_(2)O_(2)诱导HCM细胞凋亡的保护作用,同时提示AOC1是保护心肌细胞损伤治疗的潜在新靶点。Objective:To explore the molecular mechanism driving the tea polyphenol epigallocatechin-3-gallate(EGCG)-exerted protective effect on hydrogen peroxide(H_(2)O_(2))-induced cell apoptosis of human cardiomyocyte HCM cell lines.Methods:HCM cells were stimulated with H_(2)O_(2) to establish a vitro apoptosis model;the caspase-3 assay was used to assess the cell apoptosis of HCM cells;the CCK-8 assay was used to perform the cytotoxicity experiment;qPCR was used to detect the levels of AOC1 mRNA and miR-16-5p;Western blot was used to detect the protein level of AOC1;cells transfection was conducted to regulate the expression levels of AOC1 and miR-16-5p in HCM cells;TargetScan was used to predict the binding relationship and site between AOC1 mRNA 3’UTR and miR-16-5p,which was verified by the double luciferase reporter gene assay.Results:EGCG could significantly inhibit H_(2)O_(2)-induced apoptosis of HCM cells(P<0.05).In apoptotic cells,the expression of AOC1 was significantly decreased,and EGCG could significantly up-regulate the expression of AOC1,while knocking down AOC1 could significantly reverse the inhibition of EGCG on apoptosis(P<0.05).Meanwhile,the expression of miR-16-5p was significantly increased in apoptotic cells,and EGCG could significantly inhibit the expression of miR-16-5p,while up-regulation of miR-16-5p significantly reversed the inhibition of EGCG on apoptosis(P<0.05).Upregulation of miR-16-5p significantly reversed the promotion of AOC1 expression by EGCG in apoptotic cells.Finally,miR-16-5p can target inhibit the expression of AOC1,while over expression of AOC1 significantly reversed miR-16-5p-induced apoptosis of HCM cells.Conclusion:EGCG protected H_(2)O_(2)-induced apoptosis of HCM cells by regulating the miR-16-5p/AOC1 axis,which proposes AOC1 as a potential novel therapeutic target pro-tection against cardiomyocyte injury.

关 键 词：表没食子儿茶素没食子酸酯 人心肌细胞 miR-16-5p AOC1 细胞凋亡 H O 

分 类 号：TS201.1[轻工技术与工程—食品科学]