Downregulation of IRS-1 expression causes inhibition of corneal angiogenesis

Invest Ophthalmol Vis Sci. 2005 Nov;46(11):4072-8. doi: 10.1167/iovs.05-0105.

Abstract

Purpose: The antiangiogenic effect of an antisense oligodeoxynucleotide (ODN) targeting insulin receptor substrate (IRS)-1 was evaluated on rat corneal neovascularization.

Methods: Eyes with neovessels were treated with subconjunctival injections of IRS-1 antisense oligonucleotide (ASODN), IRS-1 sense ODN (SODN), or PBS. At 8 and 24 hours after the first subconjunctival injection, the expression of IRS-1, VEGF, and IL-1beta mRNA was evaluated. IRS-1 protein levels were also measured at 8 hours by Western blot analysis (n = 4/group). On day 10, corneal neovascularization was quantified in flatmount corneas of rats treated daily from days 4 to 9.

Results: On day 10, new vessels covered 95.5% +/- 4% of the corneal area in PBS-treated eyes, 92% +/- 7% in SODN-treated eyes and 59% +/- 20% in ASODN-treated eyes (P < 0.001). In the ASODN-treated group, the expression and synthesis of IRS-1 were significantly downregulated when compared with the control groups. ASODN did not significantly affect the expression of VEGF but significantly decreased the expression of IL-1beta at 24 hours (P = 0.04).

Conclusions: Subconjunctival injections of IRS-1 antisense ODN significantly inhibit rat corneal neovascularization. This effect may be mediated by a downregulation of IL-1beta. IRS-1 proteins may be interesting targets for the regulation of angiogenesis mediated by insulin, hypoxia, or inflammation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiogenesis Inhibitors / therapeutic use*
  • Animals
  • Blotting, Western
  • Conjunctiva / drug effects
  • Corneal Neovascularization / metabolism
  • Corneal Neovascularization / pathology
  • Corneal Neovascularization / prevention & control*
  • Disease Models, Animal
  • Down-Regulation
  • Immunohistochemistry
  • Injections
  • Insulin Receptor Substrate Proteins
  • Interleukin-1 / genetics
  • Interleukin-1 / metabolism
  • Oligodeoxyribonucleotides, Antisense / therapeutic use*
  • Phosphoproteins / genetics*
  • Phosphoproteins / metabolism*
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Inbred Lew
  • Specific Pathogen-Free Organisms
  • Vascular Endothelial Growth Factor A / genetics
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Angiogenesis Inhibitors
  • Insulin Receptor Substrate Proteins
  • Interleukin-1
  • Irs1 protein, rat
  • Oligodeoxyribonucleotides, Antisense
  • Phosphoproteins
  • RNA, Messenger
  • Vascular Endothelial Growth Factor A
  • vascular endothelial growth factor A, rat