Intermedin protects HUVECs from ischemia reperfusion injury via Wnt/β-catenin signaling pathway

Ren Fail. 2019 Nov;41(1):159-166. doi: 10.1080/0886022X.2019.1587468.

Abstract

Intermedin (IMD) is a member of the calcitonin gene-related peptide (CGRP) superfamily and a pro-angiogenic factor. In the present study, we identified activation of the Wnt/β-catenin signaling pathway by IMD. Adding CoCl2 HUVECs was used to establish an in vitro model. The migration of HUVECs was measured by wound healing assays and transwell migration assays. Capillary formation was measured using tube formation assays. Immunocytochemistry (ICC) analysis was used to evaluate VEGF and RAMP2 expression in HUVECs. The relevant signaling molecules were detected with western blot. Our study shows that IMD could promote H/R impaired HUVECs migration and tube formation in vitro. On the other hand, inhibition of Wnt/β-catenin signaling led to the suppression of this promotion of migration and tube formation. This result suggests that Wnt/β-catenin signaling is correlated to IMD induced angiogenesis. Analysis of results from ICC assays indicated that IMD works through increasing levels of VEGF and RAMP2. Meanwhile, the Wnt/β-catenin signaling specific inhibitor IWR-1-endo was shown to down-regulate VEGF and RAMP2 expression. Western blot results further confirmed the signaling mechanism by which IMD promotes angiogenesis. Thus, Wnt/β-catenin signaling plays an important role in IMD induced neovascularization. The data further suggest that the PI3K axis contributes positively downstream.

Keywords: IMD; Wnt/β-catenin signaling; angiogenesis; hypoxia/reoxygenation.

MeSH terms

  • Cell Line
  • Cobalt / toxicity
  • Down-Regulation / drug effects
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Imides / pharmacology
  • Neovascularization, Physiologic*
  • Peptide Hormones / metabolism*
  • Quinolines / pharmacology
  • Receptor Activity-Modifying Protein 2 / metabolism
  • Reperfusion Injury / chemically induced
  • Reperfusion Injury / pathology*
  • Up-Regulation
  • Vascular Endothelial Growth Factor A / metabolism
  • Wnt Signaling Pathway / drug effects
  • Wnt Signaling Pathway / physiology*
  • beta Catenin / metabolism

Substances

  • ADM2 protein, human
  • CTNNB1 protein, human
  • IWR-1 compound
  • Imides
  • Peptide Hormones
  • Quinolines
  • RAMP2 protein, human
  • Receptor Activity-Modifying Protein 2
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • beta Catenin
  • Cobalt
  • cobaltous chloride

Grants and funding

This work was supported by the National Nature Science Fund Project [No. 81500518, 81500529], the Youth Science and Technology Research Fund, Basic research projects in Shanxi [No. 2016021147], the Doctoral Startup Research Fund of Shanxi Medical University [No. 03201302, 03201403], the Science and Technology Innovation Fund of Shanxi Medical University [No. 01201403], and 331 Early Career Researcher Grant fund of the Basic Medical College, Shanxi Medical University [No. 201406].