SDF‑1α/CXCR4 signaling promotes capillary tube formation of human retinal vascular endothelial cells by activating ERK1/2 and PI3K pathways in vitro

Mol Med Rep. 2022 Oct;26(4):305. doi: 10.3892/mmr.2022.12821. Epub 2022 Aug 10.

Abstract

The purpose of this study is to address the effect and mechanism of stromal cell‑derived factor‑1 (SDF‑1)α/chemokine (C‑X‑C motif) receptor 4 (CXCR4) signaling on capillary tube formation of human retinal vascular endothelial cells (HRECs). The expression of CXCR4 in HRECs was quantified by reverse transcription (RT‑PCR) and western blotting. The effects of SDF‑1α/CXCR4 signaling in capillary tube formation and migration of HRECs was examined using three‑dimensional Matrigel assay and wound scratching assay respectively in vitro. Cell proliferation of HRECs was examined using cell counting kit (CCK)‑8 assay in the presence of different concentrations of SDF‑1α protein. The effect of SDF‑1α/CXCR4 signaling in HREC expression of VEGF, basic fibroblast growth factor (bFGF), IL‑8 and intercellular cell adhesion molecule (ICAM)‑1 was examined using RT‑PCR and western blotting. RT‑PCR and western blot analysis revealed CXCR4 was expressed in HRECs. The number of intact capillary tubes formed by HRECs in the presence of SDF‑1α was markedly more compared with a PBS treated control group. However, it was reduced with treatment with an CXCR4 antagonist. Wound scratching assay showed a significant increase in the number of migrated HRECs under SDF‑1α stimulation and the number was reduced with treatment with an CXCR4 antagonist. RT‑PCR and western blotting showed that SDF‑1α significantly promoted VEGF, bFGF, IL‑8 and ICAM‑1 expression in HRECs. The proliferation of HRECs in the presence of SDF‑1α was promoted in a dosage‑dependent manner. SDF‑1α/CXCR4 signaling can increase HREC capillary tube formation through promoting HREC migration, proliferation and expression of VEGF, bFGF, IL‑8 and ICAM‑1.

Keywords: chemokine; chemokine (C‑X‑C motif) receptor 4; neovascularization; retinal; stromal cell‑derived factor‑1α.

MeSH terms

  • Cell Movement
  • Cell Proliferation
  • Cells, Cultured
  • Chemokine CXCL12* / metabolism
  • Endothelial Cells* / metabolism
  • Humans
  • Intercellular Adhesion Molecule-1 / genetics
  • Intercellular Adhesion Molecule-1 / metabolism
  • Interleukin-8 / metabolism
  • MAP Kinase Signaling System
  • Phosphatidylinositol 3-Kinases / metabolism
  • Receptors, CXCR4 / metabolism
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • CXCR4 protein, human
  • Chemokine CXCL12
  • Interleukin-8
  • Receptors, CXCR4
  • Vascular Endothelial Growth Factor A
  • Intercellular Adhesion Molecule-1

Grants and funding

The present study was supported by the National Natural Science Foundation in China (grant no. 81970830), Suzhou Municipal Natural Science Foundation (grant no. SKJY2021056) and the Soochow Scholar Project of Soochow University (grant no. R5122001).