Department of Cardiology, Xiangyang Central Hospital, Affliated Hospital of Hubei College of Arts and Science, Xiangyang City, Hubei Province, 441021, China.
J Gene Med. 2021 Jan;23(1):e3274. doi: 10.1002/jgm.3274. Epub 2020 Sep 28.
Hyperglycemia increases the risk of many cardiovascular diseases (CVD), and the dysregulation of proliferation and migration in vascular smooth muscle cells (VSMCs) also participates in the pathogenesis of CVD. miR-381-3p is known to suppress the proliferation and migration of multiple human cell types. Nevertheless, the function of miR-381-3p in VSMCs remains largely indistinct.
A quantitative real-time polymerase chain reaction (qRT-PCR) was employed to investigate miR-381-3p expression in high-glucose-induced VSMCs. Inflammatory cytokines tumor necrosis factor-α, interleukin-1β and interleukin-6, as well as oxidative stress markers SOD and MDA, were determined by an enzyme-linked immunosorbent assay. Reactive oxygen species generation was examined using a 2,7'-dichlorofluorescein kit. The proliferation, migration and apoptosis of VSMCs were monitored by 3-(4,5-dimethylthiazl2-yl)-2,5-diphenyltetazolium bromide (MTT), transwell and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assays. The TargetScan database (http://www.targetscan.org) was employed to seek the potential target gene of miR-381-3p. Interaction between miR-381-3p and HMGB1 was determined by a qRT-PCR, western blotting and a luciferase reporter assay.
miR-381-3p expression was significantly reduced in a VSMCs dysfunction model induced by high-glucose in a dose- and time-dependent manner. Transfection of miR-381-3p mimics suppressed the inflammation, oxidative stress, proliferation and migration of VSMCs, whereas apoptosis of VSMCs was promoted, and the transfection of miR-381-3p inhibitors had the opposite effect. Mechanistically, HMGB1, an important factor in inflammation response, was confirmed as a target gene of miR-381-3p.
miR-381-3p targets HMGB1 to suppress the inflammation, oxidative stress, proliferation and migration of high-glucose-induced VSMCs by targeting HMGB1.
高血糖增加了许多心血管疾病（CVD）的风险，血管平滑肌细胞（VSMCs）增殖和迁移的失调也参与了 CVD 的发病机制。miR-381-3p 已知能抑制多种人类细胞类型的增殖和迁移。然而，miR-381-3p 在 VSMCs 中的功能仍很大程度上不清楚。
采用实时定量聚合酶链反应（qRT-PCR）检测高糖诱导的 VSMCs 中 miR-381-3p 的表达。采用酶联免疫吸附试验测定炎性细胞因子肿瘤坏死因子-α、白细胞介素-1β和白细胞介素-6 以及氧化应激标志物 SOD 和 MDA。采用 2'，7'-二氯荧光素试剂盒检测活性氧（ROS）的生成。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）、Transwell 和末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记（TUNEL）测定 VSMCs 的增殖、迁移和凋亡。采用 TargetScan 数据库（http://www.targetscan.org）寻找 miR-381-3p 的潜在靶基因。采用 qRT-PCR、western blot 和荧光素酶报告基因测定确定 miR-381-3p 与 HMGB1 之间的相互作用。
miR-381-3p 的表达在高糖诱导的 VSMCs 功能障碍模型中呈剂量和时间依赖性显著降低。miR-381-3p 模拟物的转染抑制了 VSMCs 的炎症、氧化应激、增殖和迁移，而促进了 VSMCs 的凋亡，而 miR-381-3p 抑制剂的转染则产生相反的效果。机制上，作为炎症反应的重要因子，HMGB1 被确认为 miR-381-3p 的靶基因。
miR-381-3p 通过靶向 HMGB1 抑制高糖诱导的 VSMCs 的炎症、氧化应激、增殖和迁移。
