Department of Infectious Diseases, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University; The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen, Guangdong, China.
J Recept Signal Transduct Res. 2021 Oct;41(5):434-441. doi: 10.1080/10799893.2020.1818095. Epub 2020 Oct 1.
MiR-145-5p is high-expressed in human vascular endothelial cells (HUVECs) and alternatively activated macrophages (M2). However, whether miR-145-5p can reduce HUVEC damage by regulating macrophage immunophenotype is less reported. THP-1 was stimulated by Phorbolate-12-myristate-13-acetate, LPS and IFN-γ, and IL-4 to differentiate into macrophages (M0, M1 and M2). The expressions of macrophage markers were detected by Western blotting, and the expressions of miR-145-5p and kruppel-like factor-14 (KLF14) were detected by qRT-PCR. Dual-luciferase reporter assay was used to analyze the targeted relationship of miR-145-5p and KLF14. HUVEC injury was induced by LPS and then co-cultured with M1 transfected by miR-145-5p mimic. The effect of miR-145-3p on proliferation and metastasis of LPS-induced HUVECs was detected by MTT, clone formation, scratch assay and Transwell. We found that the expression of miR-145-5p was higher in M2 than that in M1. MiR-145-5p expression was down-regulated during M2-to-M1, but up-regulated during M1-to-M2. The expressions of IL-1β and iNOS were down-regulated, while the protein expressions of CCL17 and Arg-1 were up-regulated by miR-145-5p mimic in M0. The viability, proliferation, migration and invasion of HUVECs were promoted, however, LDH activity of the HUVECs was inhibited by mimics. In addition, KLF14 was predicted as the target gene for miR-145-5p in HUVECs. Collectively, our results demonstrate that miR-145-5p inhibited cell proliferation of LPS-treated HUVECs possibly through regulating macrophage polarization to M2.
miR-145-5p 在人血管内皮细胞（HUVECs）和交替激活的巨噬细胞（M2）中高表达。然而，miR-145-5p 是否可以通过调节巨噬细胞免疫表型来减少 HUVEC 损伤的报道较少。THP-1 用佛波醇 12-肉豆蔻酸 13-乙酸酯、LPS 和 IFN-γ以及 IL-4 刺激分化为巨噬细胞（M0、M1 和 M2）。通过 Western blot 检测巨噬细胞标志物的表达，通过 qRT-PCR 检测 miR-145-5p 和 Kruppel 样因子 14（KLF14）的表达。双荧光素酶报告实验分析 miR-145-5p 和 KLF14 的靶向关系。用 LPS 诱导 HUVEC 损伤，然后与转染 miR-145-5p 模拟物的 M1 共培养。通过 MTT、克隆形成、划痕实验和 Transwell 检测 miR-145-3p 对 LPS 诱导的 HUVEC 增殖和转移的影响。结果发现，M2 中 miR-145-5p 的表达高于 M1。M2 向 M1 转化时 miR-145-5p 表达下调，M1 向 M2 转化时上调。M0 中 miR-145-5p 模拟物下调 IL-1β 和 iNOS 的表达，上调 CCL17 和 Arg-1 的蛋白表达。Mimics 促进 HUVEC 的活力、增殖、迁移和侵袭，同时抑制 HUVEC 的 LDH 活性。此外，在 HUVEC 中预测 KLF14 是 miR-145-5p 的靶基因。综上所述，我们的研究结果表明，miR-145-5p 通过调节巨噬细胞向 M2 极化抑制 LPS 处理的 HUVEC 细胞增殖。
