Yang Yingjie, Ou Yanchun, Mo Guanlian, Wen Jing, Liang Limin, Wang Shirong, Li Jinyi
Guilin Medical University Affiliated Hospital, Guilin, Guangxi, China.
PLoS One. 2025 Sep 18;20(9):e0332892. doi: 10.1371/journal.pone.0332892. eCollection 2025.
Heart failure (HF) is a major cardiovascular disease with high mortality worldwide, whose pathophysiology is multifaceted. Hypoxia has emerged as a critical factor contributing to the progression of heart failure. We aimed to examine the expression and functions of LncRNA Kcnq1ot1 in hypoxia-induced cardiomyocytes in the process of HF.
H9C2 cell model was simulated by hypoxia treatment. TUNEL, ELISA, Western Blot and qRT-PCR assay were carried out to evaluate cell pyroptosis, inflammation and dysfunction. Subsequently, we identified the direct downstream target of Kcnq1ot1 by bioinformatics analysis, RNA pull-down, double Luciferase reporter gene and other functional experiments.
Firstly, Kcnq1ot1 levels was revealed to be upregulated in hypoxia cells than in control cells, and miR-27b-3p showed the opposite trend. And as expected, inhibition of Kcnq1ot1 and overexpression of miR-27b-3p both protected H9C2 against hypoxia-induced pyroptosis, inflammation and dysfunction. Moreover, miR-27b-3p was proved to bind with Kcnq1ot1 and participated in Kcnq1ot1-mediated H9C2 injury under hypoxia by regulating the Wnt3a/β-Catenin/NLRP3 signaling pathway.
Collectively, our study demonstrated that inhibition of Kcnq1ot1 protected cardiomyocyte against hypoxia-induced injury possibly via sponging miR-27b-3p, which could be useful as biomarkers and therapeutic targets for HF patients.
心力衰竭（HF）是一种主要的心血管疾病，在全球范围内死亡率很高，其病理生理学是多方面的。缺氧已成为导致心力衰竭进展的关键因素。我们旨在研究长链非编码RNA Kcnq1ot1在心力衰竭过程中缺氧诱导的心肌细胞中的表达及功能。
通过缺氧处理模拟H9C2细胞模型。采用TUNEL、ELISA、蛋白质免疫印迹法和qRT-PCR检测法评估细胞焦亡、炎症反应及功能障碍。随后，通过生物信息学分析、RNA下拉实验、双荧光素酶报告基因等功能实验鉴定Kcnq1ot1的直接下游靶点。
首先，与对照细胞相比，缺氧细胞中Kcnq1ot1水平上调，而miR-27b-3p呈现相反趋势。正如预期的那样，抑制Kcnq1ot1和过表达miR-27b-3p均可保护H9C2细胞免受缺氧诱导的焦亡、炎症反应及功能障碍。此外，miR-27b-3p被证明与Kcnq1ot1结合，并通过调节Wnt3a/β-连环蛋白/NLRP3信号通路参与缺氧条件下Kcnq1ot1介导的H9C2细胞损伤。
总的来说，我们的研究表明，抑制Kcnq1ot1可能通过海绵吸附miR-27b-3p保护心肌细胞免受缺氧诱导的损伤，这可能作为HF患者的生物标志物和治疗靶点。
