沉默长链非编码 RNA KCNQ1OT1 通过 miR-377-3p/HMOX1 减轻心肌缺血/再灌注损伤（MI/RI）诱导的心脏损伤的机制。
Mechanisms by which silencing long-stranded noncoding RNA KCNQ1OT1 alleviates myocardial ischemia/reperfusion injury (MI/RI)-induced cardiac injury via miR-377-3p/HMOX1.
机构信息
Center for Rehabilitation Medicine, Rehabilitation & Sports Medicine Research Institute of Zhejiang Province, Department of Rehabilitation Medicine, Zhejiang Provincial People's Hospital (Affiliated People's Hospital, Hangzhou Medical College, Hangzhou, Zhejiang, 310014, China.
Medical Experimental Center, The First Affiliated Hospital of Chongqing Medical and Pharmaceutical College (The 6th People Hospital of Chongqing), Chongqing, 400060, China.
出版信息
BMC Cardiovasc Disord. 2024 Jan 3;24(1):19. doi: 10.1186/s12872-023-03693-y.
BACKGROUND
The key complication of myocardial infarction therapy is myocardial ischemia/reperfusion injury (MI/RI), and there is no effective treatment. The present study elucidates the mechanism of action of lncRNA KCNQ1OT1 in alleviating MI/RI and provides new perspectives and therapeutic targets for cardiac injury-related diseases.
METHODS
An ischemia/reperfusion (I/R) injury model of human adult cardiac myocytes (HACMs) was constructed, and the expression of KCNQ1OT1 and miR-377-3p was determined by RT‒qPCR. The levels of related proteins were detected by western blot analysis. Cell proliferation was detected by a CCK-8 assay, and cell apoptosis and ROS content were determined by flow cytometry. SOD and MDA expression as well as Fe changes were detected by related analysis kits. The target binding relationships between lncRNA KCNQ1OT1 and miR-377-3p as well as between miR-377-3p and heme oxygenase 1 (HMOX1) were verified by a dual-luciferase reporter gene assay.
RESULTS
Myocardial ischemia‒reperfusion caused oxidative stress in HACMs, resulting in elevated ROS levels, increased Fe levels, decreased cell viability, and increased LDH release (a marker of myocardial injury), and apoptosis. KCNQ1OT1 and HMOX1 were upregulated in I/R-induced myocardial injury, but the level of miR-377-3p was decreased. A dual-luciferase reporter gene assay indicated that lncRNA KCNQ1OT1 targets miR-377-3p and that miR-377-3p targets HMOX1. Inhibition of HMOX1 alleviated miR-377-3p downregulation-induced myocardial injury. Furthermore, lncRNA KCNQ1OT1 promoted the level of HMOX1 by binding to miR-377-3p and aggravated myocardial injury.
CONCLUSION
LncRNA KCNQ1OT1 aggravates ischemia‒reperfusion-induced cardiac injury via miR-377-3P/HMOX1.
背景
心肌梗死治疗的关键并发症是心肌缺血/再灌注损伤（MI/RI），目前尚无有效的治疗方法。本研究阐明了长链非编码 RNA KCNQ1OT1 缓解 MI/RI 的作用机制，为心脏损伤相关疾病提供了新的视角和治疗靶点。
方法
构建人成年心肌细胞（HACMs）缺血/再灌注（I/R）损伤模型，通过 RT‒qPCR 检测 KCNQ1OT1 和 miR-377-3p 的表达。通过 Western blot 分析检测相关蛋白水平。通过 CCK-8 检测细胞增殖，通过流式细胞术检测细胞凋亡和 ROS 含量。通过相关分析试剂盒检测 SOD 和 MDA 的表达以及 Fe 的变化。通过双荧光素酶报告基因实验验证 lncRNA KCNQ1OT1 与 miR-377-3p 以及 miR-377-3p 与血红素加氧酶 1（HMOX1）之间的靶标结合关系。
结果
心肌缺血再灌注导致 HACMs 发生氧化应激，导致 ROS 水平升高、Fe 水平升高、细胞活力降低、LDH 释放（心肌损伤标志物）增加和凋亡增加。KCNQ1OT1 和 HMOX1 在 I/R 诱导的心肌损伤中上调，而 miR-377-3p 的水平下降。双荧光素酶报告基因实验表明，lncRNA KCNQ1OT1 靶向 miR-377-3p，miR-377-3p 靶向 HMOX1。抑制 HMOX1 减轻了 miR-377-3p 下调诱导的心肌损伤。此外，lncRNA KCNQ1OT1 通过与 miR-377-3p 结合促进 HMOX1 的水平，加重心肌损伤。
结论
lncRNA KCNQ1OT1 通过 miR-377-3P/HMOX1 加重缺血/再灌注诱导的心脏损伤。
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