Up-regulation of MiR-145-5p promotes the growth and migration in LPS-treated HUVECs through inducing macrophage polarization to M2

Abstract

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.

Keywords:

• MiR-145-5p
• macrophage
• human vascular endothelial cells
• lipopolysaccharide
• kruppel-like factor-13

Acknowledgements

Thanks for useful suggestions given by Dr Sam Xu of Hull University in UK.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

The present study was supported by the Shenzhen Municipal Science and Technology Innovation Committee Project [JCYJ 20140416122812003].