The Graduate School, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China.
Eur Rev Med Pharmacol Sci. 2020 Oct;24(20):10829-10838. doi: 10.26355/eurrev_202010_23445.
This study aims to investigate the role of miR-34a-5p in the regulation of lipopolysaccharide (LPS)-induced injury of vascular endothelial cells (ECs).
Human umbilical vein ECs (HUVECs) were exposed to LPS to stimulate endothelial injury in vitro. miRNA microarray analysis was carried out to identify miR-34a-5p expression in HUVECs. MTT and flow cytometry analyses were used to determine cell viability and apoptosis rate, respectively. Furthermore, enzyme-linked immunosorbent assay (ELISA), qRT-PCR, and Western blot were used to examine the factors involved in inflammation and their relative gene expression. Additionally, Matrigel-based tube formation assay was carried out to assess the vasculogenic activity of HUVECs. Luciferase reporter assay was used to analyze the possible relationship between miR-34a-5p and FOXM1.
MiR-34a-5p expression was significantly enhanced in HUVECs after 24 h of LPS treatment. LPS treatment led to a dramatic inhibition of cell viability, enhanced apoptosis, increased production of pro-inflammatory cytokines, and inhibited the vasculogenic activity of HUVECs. MiR-34a-5p inhibitor attenuated LPS-induced damage. MiR-34a-5p directly inhibited the expression of FOXM1, and its overexpression alleviated the protective effect of FOXM1 on cell viability, apoptosis, inflammation factor production, and vasculogenic activity. The activity of the NRF2/HO-1 pathway was inhibited by miR-34a-5p, possibly via FOXM1.
MiR-34a-5p inhibition attenuates LPS-induced EC injury by targeting FOXM1 via activation of the NRF2/HO-1 pathway.
本研究旨在探讨 miR-34a-5p 在脂多糖（LPS）诱导的血管内皮细胞（EC）损伤中的作用。
体外采用 LPS 刺激人脐静脉内皮细胞（HUVEC）以模拟内皮损伤。通过 miRNA 微阵列分析鉴定 HUVEC 中 miR-34a-5p 的表达。MTT 和流式细胞术分别用于检测细胞活力和凋亡率。此外，酶联免疫吸附试验（ELISA）、qRT-PCR 和 Western blot 用于检测炎症相关因子及其相对基因表达。另外，采用 Matrigel 基管形成实验评估 HUVEC 的血管生成活性。荧光素酶报告实验分析 miR-34a-5p 与 FOXM1 之间的可能关系。
LPS 处理 24 h 后 HUVEC 中 miR-34a-5p 表达明显增强。LPS 处理导致细胞活力显著抑制，凋亡增加，促炎细胞因子产生增加，并抑制 HUVEC 的血管生成活性。miR-34a-5p 抑制剂减弱了 LPS 诱导的损伤。miR-34a-5p 可直接抑制 FOXM1 的表达，而过表达 FOXM1 可减轻 FOXM1 对细胞活力、凋亡、炎症因子产生和血管生成活性的保护作用。miR-34a-5p 抑制 NRF2/HO-1 通路的活性，可能通过 FOXM1。
miR-34a-5p 通过靶向 FOXM1 抑制 LPS 诱导的 EC 损伤，激活 NRF2/HO-1 通路。
