Department of Cardiac Function, The Affiliated Hospital of Kangda College of Nanjing Medica University/The First People's Hospital of Lianyungang/ The Affiliated Lianyungang Hospital of Xuzhou Medical University, Lianyungang, Jiangsu, China (mainland).
Department of Cardiac Surgery, First hospital of Jilin University, Changchun, Jilin, China (mainland).
Med Sci Monit. 2020 Apr 24;26:e921877. doi: 10.12659/MSM.921877.
BACKGROUND Endothelial cell (EC) injury is underlies for the pathogenesis of atherosclerosis (AS). MicroRNAs (miRNAs) have been indicated play important role in modulating AS occurrence and development. However, how miR-328-3p modulates EC injury molecular level for AS remains unclear. MATERIAL AND METHODS MiR-328-3p and forkhead box protein O4 (FOXO4) expression were detected using quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. Cell viability was analyzed by Cell Counting Kit-8 (CCK-8) assay. Flow cytometry was utilized to analyze the apoptotic rate. The migration and invasion abilities were measured by Transwell assay. Western blot was applied to examine the expression of C-caspase 3, Beclin, LC3-I, and LC3-II. The levels of interleukin (IL)-1ß, IL-10, and tumor necrosis factor-alpha (TNF-alpha) were tested by enzyme-linked immunosorbent assay (ELISA) assay and western blot. RESULTS MiR-328-3p expression was downregulated in oxidized low-density lipoprotein (ox-LDL) induced human umbilical vein endothelial cells (HUVECs). Overexpressed miR-328-3p obviously alleviated ox-LDL induced inhibition on cell viability, migration and invasion, stimulation on apoptosis, autophagy as well as inflammation in HUVECs. FOXO4 was elevated in ox-LDL HUVECs, and functional assay indicated that FOXO4 aggravated ox-LDL induced HUVECs impairment. In addition, FOXO4 was a target of miR-328-3p in HUVECs; rescue experiments suggested miR-328-3p could protect HUVECs against ox-LDL induced injury via regulating FOXO4. CONCLUSIONS MiR-328-3p protected vascular endothelial cells against ox-LDL induced injury via targeting FOXO4, suggesting a novel insight for atherosclerosis treatment.
血管内皮细胞（EC）损伤是动脉粥样硬化（AS）发病机制的基础。microRNAs（miRNAs）已被证明在调节 AS 的发生和发展中发挥重要作用。然而，miR-328-3p 如何调节 EC 损伤的分子水平尚不清楚。
采用实时定量聚合酶链反应（qRT-PCR）和 Western blot 检测 miR-328-3p 和叉头框蛋白 O4（FOXO4）的表达。细胞活力通过细胞计数试剂盒-8（CCK-8）测定。流式细胞术用于分析细胞凋亡率。Transwell 测定用于检测迁移和侵袭能力。Western blot 用于检测 C-caspase 3、Beclin、LC3-I 和 LC3-II 的表达。酶联免疫吸附试验（ELISA）和 Western blot 检测白细胞介素（IL）-1β、IL-10 和肿瘤坏死因子-α（TNF-α）的水平。
氧化低密度脂蛋白（ox-LDL）诱导的人脐静脉内皮细胞（HUVECs）中 miR-328-3p 表达下调。过表达 miR-328-3p 明显减轻 ox-LDL 诱导的 HUVECs 活力抑制、迁移和侵袭刺激、凋亡、自噬和炎症。ox-LDL HUVECs 中 FOXO4 升高，功能测定表明 FOXO4 加重 ox-LDL 诱导的 HUVECs 损伤。此外，FOXO4 是 HUVECs 中 miR-328-3p 的靶标；挽救实验表明，miR-328-3p 可通过调节 FOXO4 保护 HUVECs 免受 ox-LDL 诱导的损伤。
miR-328-3p 通过靶向 FOXO4 保护血管内皮细胞免受 ox-LDL 诱导的损伤，为动脉粥样硬化的治疗提供了新的思路。
