首页|内皮祖细胞微粒miR-126调控线粒体改善心肌细胞OGD/R损伤的机制研究

内皮祖细胞微粒miR-126调控线粒体改善心肌细胞OGD/R损伤的机制研究

Study on mechanism of endothelial progenitor cell particle miR-126 in regulating mitochondria to improve OGD/R damage in cardiomyocytes

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目的 研究微小RNA-126(miR-126)-内皮祖细胞(EPC)-微粒(MPs)改善心肌细胞氧糖剥夺/复氧(OGD/R)损伤的分子机制.方法 采用心肌细胞建立对照(control)和OGD/R损伤模型,并分别给予EPC-MPs和miR-126 mimic EPC-MPs处理.通过透射电子显微镜评估心肌细胞细胞器的结构变化.采用ELISA检测心肌细胞中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)和高迁移率族蛋白B1(HMGB1)的表达水平.采用Western blot和实时荧光定量PCR(qPCR)检测不同处理后心肌细胞中血管紧张素Ⅱ(AngⅡ)、半胱氨酸天冬氨酸蛋白酶-6(caspase-6)、一氧化氮合酶(eNOS)、叉头框蛋白O1(FOXO1)、髓过氧化物酶(MPO)、核因子-κB(NF-κB)、细胞外信号调节激酶1(ERK1)、肝激酶B1(LKB1)、沉默信息调节因子2相关酶1(SIRT1)、葡萄糖调节蛋白78(GRP78)、130×103高尔基体基质蛋白(GM130)和过氧化物酶体增生物激活受体γ共激活因子-1α(PGC-1α)的蛋白和mRNA表达水平.结果 OGD/R+miR-126 mimic EPC-MPs组部分心肌细胞线粒体结构完整,部分线粒体大小增大,内质网表面核糖体较少,高尔基体部分囊泡聚集.与OGD/R+EPC-MPs组比较,OGD/R+miR-126 mimic EPC-MPs组IL-6、TNF-α和HMGB-1的表达水平下调.与OGD/R+EPC-MPs组比较,OGD/R+miR-126 mimic EPC-MPs组AngⅡ、caspase-6、p-ERK1/2和GRP78的蛋白表达水平下调,p-LKB1、GM130和PGC-1α的蛋白表达水平上调.qPCR结果显示,与OGD/R+EPC-MPs组比较,OGD/R+miR-126 mimic EPC-MPs组AngⅡ、MPO和GRP78的mRNA表达水平下调,eNOS、GM130和PGC-1α的mRNA表达水平上调.结论 当心肌细胞发生OGD/R损伤时,miR-126 mimic EPC-MPs调节PGC-1α/GM130表达,抑制AngⅡ诱导的应激损伤反应,减少心肌细胞的OGD/R损伤.
Objective To investigate the molecular mechanism of miR-126-EPC-MPs in ameliorating oxygen glucose deprivation/reoxygenation (OGD/R) injury in cardiomyocytes.Methods The cardiomyocytes were used to establish the control and OGD/R injury models,and EPC-MPs and miR-126 mimic EPC-MPs treatments were administered respectively.The structural changes in cardiomyocyte organelles were assessed by transmission electron microscopy.ELISA was used to detect the expression levels of IL-6,TNF-α and HMGB-1 in cardiomyocytes.The protein and mRNA expression levels of Ang Ⅱ,caspase-6,eNOS,FOXO1,MPO,NF-κB,ERK1,LKB1,SIRT1,GRP78,GM130 and PGC-1α in cardiomyocytes after different treatments were detected by Western blot and quantitative PCR.Results The mitochondria structure in some cardiomyo-cytes of the OGD/R+miR-126 mimic EPC-MPs group was complete,the size of some mitochondria was in-creased,the surface of the endoplasmic reticulum had fewer ribosomes,and some of the Golgi vesicles gath-ered.Compared with the OGD/R+EPC-MPs group,the expression levels of IL-6,TNF-α and HMGB-1 in the OGD/R+miR-126 mimic EPC-MPs group were down-regulated.Compared with the OGD/R+EPC-MPs group,the expression levels of Ang Ⅱ,caspase-6,p-ERK1/2 and GRP78 protein in the OGD/R+miR-126 mimic EPC-MPs group were down-regulated,and the expression levels of p-LKB1,GM130 and PGC-1α protein in the OGD/R+miR-126 mimic EPC-MPs group were up-regulated.The quantitative PCR(qPCR) results showed that compared with the OGD/R+EPC-MPs group,the expression levels of Ang Ⅱ,MPO and GRP78 in the OGD/R+miR-126 mimic EPC-MPs group were down-regulated,and the expression levels of eNOS,GM130 and PGC-1α were up-regulated.Conclusion When OGD/R injury occurs in cardiomyocytes,miR-126 mimic EPC-MPs regulates PGC-1α/GM130 expression and inhibits Ang Ⅱ-induced stress injury response,thereby reduces the OGD/R damage in cardiomyocytes.

endothelial progenitor cellsmicroparticlesmiR-126mitochondriaacute myocardial in-farctioncardiomyocytesischemia-reperfusion injury

马艺萍、阿卜拉江·艾合麦提、帕丽达·玉山江、刘柯、卡迪尔亚·依布拉音、麦迪乃姆·阿卜力孜、穆叶赛·尼加提

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新疆医科大学研究生院,乌鲁木齐 830017

新疆维吾尔自治区人民医院急救中心,乌鲁木齐 830001

内皮祖细胞 微粒 miR-126 线粒体 急性心肌梗死 心肌细胞 缺血再灌注损伤

国家自然科学基金地区基金项目国家自然科学基金地区基金项目新疆维吾尔自治区研究生创新项目

8206007682360068XJ2023G202

2024

重庆医学
重庆市卫生信息中心,重庆市医学会

重庆医学

CSTPCD
影响因子:1.797
ISSN:1671-8348
年,卷(期):2024.53(11)