Department of Critical Care Medicine, Affiliated Haikou Hospital of Xiangya Medical College, Central South University.
Department of Pulmonary and Critical Care Medicine, Hainan General Hospital.
Int Heart J. 2021 May 29;62(3):636-646. doi: 10.1536/ihj.20-512. Epub 2021 May 15.
Sepsis-induced myocardial dysfunction (SIMD) is a common complication with high incidence rates in sepsis patients. This study aimed to investigate the roles of miR-210-3p in regulating cardiomyocyte apoptosis and mitochondrial dysfunction associated with SIMD pathogenesis.A rat sepsis model was established by cecal ligation and puncture. Serum inflammatory factors, myocardial tissue apoptosis, and expression of miR-210-3p were evaluated. In vitro, miR-210-3p expression in H9C2 cells was altered by transfection with its mimics or inhibitors. H9C2 viability was assessed via CCK-8 assay, and reactive oxygen species (ROS) production and apoptosis were detected through flow cytometry. The targeting regulatory relations between miR-210-3p and NADH dehydrogenase (ubiquinone) 1 alpha subcomplex 4 (NDUFA4) were validated by dual luciferase reporter assay.The rat sepsis model showed increased serum TNF-α and IL-6 levels, significant myocardial tissue injuries and apoptosis with decreased Bcl-2 and increased Caspase-1 protein levels. In vitro, septic rat serum suppressed viability, promoted ROS production and apoptosis, impaired COX IV activities and increased cytochrome release in H9C2 cells. The expression of miR-210-3p was greatly increased in myocardial tissues of septic rats and septic serum-treated H9C2 cells. miR-210-3p directly binds to the 3' UTR of the NDUFA4 gene. Septic rat serum suppressed NDUFA4 and Iron-Sulfur Cluster Assembly Protein U gene expressions in H9C2 cells. The above cellular and molecular alterations in H9C2 cells induced by septic serum were enhanced by miR-210-3p mimics and abrogated by miR-210-3p inhibitors.miR-210-3p promoted SIMD pathogenesis by targeting NDUFA4 to enhance cardiomyocyte apoptosis and impair mitochondrial function.
脓毒症诱导性心肌功能障碍（SIMD）是脓毒症患者中一种常见的高发病率并发症。本研究旨在探讨 miR-210-3p 在调节与 SIMD 发病机制相关的心肌细胞凋亡和线粒体功能障碍中的作用。通过盲肠结扎和穿孔建立大鼠脓毒症模型。评估血清炎症因子、心肌组织凋亡和 miR-210-3p 的表达。在体外，通过转染其模拟物或抑制剂改变 H9C2 细胞中 miR-210-3p 的表达。通过 CCK-8 测定评估 H9C2 细胞活力，并通过流式细胞术检测活性氧（ROS）产生和细胞凋亡。通过双荧光素酶报告基因检测验证 miR-210-3p 与烟酰胺腺嘌呤二核苷酸脱氢酶（泛醌）1 亚基复合物 4（NDUFA4）之间的靶向调节关系。
大鼠脓毒症模型显示血清 TNF-α和 IL-6 水平升高，心肌组织损伤和凋亡明显，Bcl-2 降低，Caspase-1 蛋白水平升高。在体外，脓毒症大鼠血清抑制 H9C2 细胞活力，促进 ROS 产生和凋亡，损害 COX IV 活性并增加细胞色素释放。脓毒症大鼠心肌组织和脓毒症血清处理的 H9C2 细胞中 miR-210-3p 的表达大大增加。miR-210-3p 直接与 NDUFA4 基因的 3'UTR 结合。脓毒症大鼠血清抑制 H9C2 细胞中 NDUFA4 和 Iron-Sulfur Cluster Assembly Protein U 基因的表达。脓毒症血清诱导的 H9C2 细胞的上述细胞和分子改变，被 miR-210-3p 模拟物增强，被 miR-210-3p 抑制剂减弱。
miR-210-3p 通过靶向 NDUFA4 促进 SIMD 发病机制，增强心肌细胞凋亡并损害线粒体功能。
