Department of Anesthesiology, Wuhan Fourth Hospital, Wuhan, Hubei, China.
Bioengineered. 2022 May;13(5):13300-13311. doi: 10.1080/21655979.2022.2080420.
Dexmedetomidine (Dex) protects the heart from ischemia/reperfusion (I/R) injury. The differential expression of long non-coding RNAs (lncRNAs) is associated with myocardial injury, but whether the lncRNA colon cancer-associated transcript 1 (CCAT1) is associated with Dex-mediated myocardial protection remains unclear. In this study, a hypoxia/reoxygenation (H/R) H9C2 model was established to simulate the in vitro characteristics of I/R. CCAT1 and microRNA (miR)-8063 expression levels in H/R H9C2 cells pretreated with Dex were determined via quantitative reverse transcription-polymerase chain reaction. The survival and apoptotic rates of H9C2 cells were determined via cell counting kit-8 and flow cytometry assays. Wnt3a, Wnt5a, and β-catenin protein levels were measured via western blotting. Luciferase and RNA immunoprecipitation assays were used to explore the binding relationship between miR-8063 and CCAT1. Dex pretreatment increased H/R H9C2 cell viability and CCAT1 expression, while decreasing the cell apoptosis and Wnt3a, Wnt5a, and β-catenin protein levels. Knockdown of CCAT1 abolished the protective effects of Dex on H/R H9C2 cells, and the downregulation of miR-8063 expression eliminated the effect of CCAT1 knockdown. These results revealed that CCAT1, a sponge for miR-8063, is involved in Dex-mediated H9C2 cell H/R injury by negatively targeting miR-8063 and inactivating the Wnt/β-catenin pathway. Dex protects H9C2 cells from H/R impairment by regulating the lncRNA CCAT1/miR-8063/Wnt/β-catenin axis.
右美托咪定（Dex）可保护心脏免受缺血/再灌注（I/R）损伤。长链非编码 RNA（lncRNA）的差异表达与心肌损伤有关，但 lncRNA 结肠癌相关转录物 1（CCAT1）是否与 Dex 介导的心肌保护有关尚不清楚。本研究通过建立缺氧/复氧（H/R）H9C2 模型，模拟体外 I/R 特征。通过实时定量逆转录聚合酶链反应检测 Dex 预处理的 H/R H9C2 细胞中 CCAT1 和 microRNA（miR）-8063 的表达水平。通过细胞计数试剂盒-8 和流式细胞术检测 H9C2 细胞的存活率和凋亡率。通过 Western blot 检测 Wnt3a、Wnt5a 和β-catenin 蛋白水平。通过荧光素酶和 RNA 免疫沉淀试验探讨 miR-8063 与 CCAT1 的结合关系。Dex 预处理可增加 H/R H9C2 细胞活力和 CCAT1 表达，降低细胞凋亡和 Wnt3a、Wnt5a 和β-catenin 蛋白水平。CCAT1 敲低可消除 Dex 对 H/R H9C2 细胞的保护作用，而 miR-8063 表达下调则消除了 CCAT1 敲低的作用。这些结果表明，作为 miR-8063 海绵的 CCAT1 通过负向靶向 miR-8063 并使 Wnt/β-catenin 通路失活，参与 Dex 介导的 H9C2 细胞 H/R 损伤。Dex 通过调节 lncRNA CCAT1/miR-8063/Wnt/β-catenin 轴来保护 H9C2 细胞免受 H/R 损伤。
