Liu Z, Cao S, Chen Q, Fu F, Cheng M, Huang X
Dermatology Hospital of Southern Medical University, Guangzhou 510030, China.
Department of Thoracic Surgery, Southern Medical University, Guangzhou 510515, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2022 Jan 20;42(1):143-149. doi: 10.12122/j.issn.1673-4254.2022.01.18.
To explore the expression of microRNA-132 (miR-132) and its potential role in the development of atherosclerosis (AS).
Thirty AS samples and 30 samples of normal peripheral vessels were collected from atherosclerotic patients undergoing peripheral angiostomy in our hospital for detecting the expression level of miR-132 using RT-qPCR. The expression of miR-132 in human umbilical vein endothelial cells (HUVEC) was up-regulated by liposome transfection, and intracellular reactive oxygen species (ROS), localization relationship between ROS and mitochondria, functional changes of mitochondrial reactive oxygen superoxide species (mtROS), mitochondrial membrane potential (MMP) and opening of mitochondrial permeability transition pore (mPTP) were analyzed by flow cytometry and laser confocal microscopy. The activity of mitochondrial redox respiratory chain complex (type I, II, III, IV and V) in HUVECs was detected using ELISA, and the expression levels of key iron death proteins were detected with Western blotting.
RT-qPCR results showed that miR-132 was significantly up-regulated in atherosclerotic plaques compared with normal vascular samples ( < 0.001). Compared with control HUVECs, HUVECs overexpressing miR-132 showed a significantly increased level of intracellular ROS ( < 0.001), and most of ROS was colocalized with mitochondria. HUVECs overexpressing miR-132 also showed significantly decreased MMP ( < 0.001) and obviously increased mtROS ( < 0.001) and opening of mPTP ( < 0.001), which led to mitochondrial REDOX respiratory chain stress disorder. The key iron death protein GPX4 was significantly down-regulated and the oxidized protein NOX4 was significantly increased in miR-132-overexpressing HUVECs ( < 0.001).
MiR-132 promotes atherosclerosis by inducing mitochondrial oxidative stress-mediated ferroptosis, which may serve as a promising therapeutic target for AS.
探讨微小RNA-132（miR-132）的表达及其在动脉粥样硬化（AS）发生发展中的潜在作用。
收集我院行外周血管造影术的动脉粥样硬化患者的30例AS样本和30例正常外周血管样本，采用RT-qPCR检测miR-132的表达水平。通过脂质体转染上调人脐静脉内皮细胞（HUVEC）中miR-132的表达，采用流式细胞术和激光共聚焦显微镜分析细胞内活性氧（ROS）、ROS与线粒体的定位关系、线粒体活性氧超氧化物（mtROS）的功能变化、线粒体膜电位（MMP）及线粒体通透性转换孔（mPTP）的开放情况。采用ELISA检测HUVEC中线粒体氧化还原呼吸链复合物（I、II、III、IV和V型）的活性，采用蛋白质免疫印迹法检测关键铁死亡蛋白的表达水平。
RT-qPCR结果显示，与正常血管样本相比，动脉粥样硬化斑块中miR-132显著上调（<0.001）。与对照HUVEC相比，过表达miR-132的HUVEC细胞内ROS水平显著升高（<0.001），且大部分ROS与线粒体共定位。过表达miR-132的HUVEC还表现出MMP显著降低（<0.001）、mtROS明显升高（<0.001）及mPTP开放增加（<0.001），导致线粒体氧化还原呼吸链应激紊乱。在过表达miR-132的HUVEC中，关键铁死亡蛋白GPX4显著下调，氧化蛋白NOX4显著升高（<0.001）。
MiR-132通过诱导线粒体氧化应激介导的铁死亡促进动脉粥样硬化，这可能为AS提供一个有前景的治疗靶点。
