Interleukin-1beta augments angiogenic responses of murine endothelial progenitor cells in vitro

J Cereb Blood Flow Metab. 2009 May;29(5):933-43. doi: 10.1038/jcbfm.2009.17. Epub 2009 Feb 25.

Abstract

Endothelial progenitor cells (EPCs) may provide novel opportunities for therapeutic angiogenesis after ischemic diseases. However, it is unclear how the angiogenic potential of EPCs might be affected by an inflammatory environment. We examine how the potent cytokine interleukin-1beta (IL-1beta) affects angiovasculogenic responses in EPCs in culture. Mononuclear cells isolated from mouse spleen were plated on fibronectin-coated wells and grown in EGM-2 MV media. Endothelial progenitor cells were phenotyped using multiple markers (UEA-Lectin, ac-LDL, CD133, CD34, vWillebrand Factor, Flk-1) and to identify the IL-1 Receptor-I. We quantified cell and colony counts and performed MTT (3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide) and Matrigel assays, in vitro, under control and IL-1beta (10 ng/mL) conditions. Endothelial progenitor cells exposed to IL-1beta increased in the number of cells and colonies compared with untreated cells, without any effect on cell metabolic integrity. Furthermore, IL-1beta treatment augmented EPC angiogenic function, significantly increasing the number of vessel-like structures in the Matrigel assay. An early phosphorylation of ERK1/2 occurred after IL-1beta stimulation, and this pathway was inhibited if IL-1 Receptor-I was blocked. Our results suggest that IL-1beta is a potent stimulator of in vitro angiogenesis through ERK signaling in mouse EPCs. Further studies are warranted to assess how interactions between proinflammatory environments and EPC responses may be leveraged to enhance therapeutic angiogenesis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • Cell Culture Techniques
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Endothelial Cells / cytology
  • Endothelial Cells / drug effects*
  • Endothelial Cells / enzymology
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Interleukin-1beta / pharmacology*
  • Interleukin-1beta / physiology
  • Male
  • Mice
  • Mice, Inbred Strains
  • Neovascularization, Physiologic / drug effects*
  • Spleen / cytology
  • Stem Cells / cytology
  • Stem Cells / drug effects*
  • Stem Cells / enzymology

Substances

  • Interleukin-1beta
  • Extracellular Signal-Regulated MAP Kinases