Department of Stem Cells and Regenerative Medicine, College of Basic Medical Science, China Medical University, Shenyang 110122, China.
Department of Anesthesiology, First Affiliated Hospital of China Medical University, Shenyang 110001, China.
Biomed Res Int. 2020 Mar 3;2020:8150780. doi: 10.1155/2020/8150780. eCollection 2020.
Mesenchymal stem cells (MSCs) have been a potential strategy in the pretreatment of pulmonary diseases, while the mechanisms of MSCs-conditioned medium (MSCs-CM) involved with microRNAs on the regulation of lung ion transport are seldom reported. We investigated the role of miR-124-5p in lipopolysaccharide-involved epithelial sodium channel (ENaC) dysfunction and explored the potential target of miR-124-5p. We observed the lower expression of miR-124-5p after the administration of MSCs-CM, and the overexpression or inhibition of miR-124-5p regulated epithelial sodium channel -subunit (-ENaC) expression at protein levels in mouse alveolar type 2 epithelial (AT2) cells. We confirmed that -ENaC is one of the target genes of miR-124-5p through dual luciferase assay and Ussing chamber assay revealed that miR-124-5p inhibited amiloride-sensitive currents associated with ENaC activity in intact H441 monolayers. Our results demonstrate that miR-124-5p can decrease the expression and function of -ENaC in alveolar epithelial cells by targeting the 3'-UTR. The involvement of MSCs-CM in lipopolysaccharide-induced acute lung injury cell model could be related to the downregulation of miR-124-5p on -ENaC, which may provide a new target for the treatment of acute lung injury.
间充质干细胞 (MSCs) 已成为肺部疾病预处理的一种有潜力的策略，而 MSCs 条件培养基 (MSCs-CM) 中涉及 microRNAs 调节肺离子转运的机制则很少有报道。我们研究了 miR-124-5p 在脂多糖参与的上皮钠通道 (ENaC) 功能障碍中的作用，并探索了 miR-124-5p 的潜在靶标。我们观察到 MSCs-CM 给药后 miR-124-5p 的表达降低，并且在小鼠肺泡型 2 上皮 (AT2) 细胞中，miR-124-5p 的过表达或抑制调节上皮钠通道 -亚基 (-ENaC) 的蛋白水平表达。我们通过双荧光素酶测定证实 -ENaC 是 miR-124-5p 的靶基因之一，Ussing 室测定显示 miR-124-5p 抑制了完整 H441 单层中与 ENaC 活性相关的氨苯蝶啶敏感电流。我们的结果表明，miR-124-5p 可以通过靶向 3'-UTR 降低肺泡上皮细胞中 -ENaC 的表达和功能。MSCs-CM 在脂多糖诱导的急性肺损伤细胞模型中的参与可能与 miR-124-5p 下调 -ENaC 有关，这可能为急性肺损伤的治疗提供新的靶标。
