May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Association Between High Glucose-Induced Overexpression of Collagen Type IV and Integrin Expression in Microvascular Endothelial Cells
Author Affiliations & Notes
  • M. Wong
    Departments of Medicine and Ophthalmology, Boston University School of Medicine, Boston, Massachusetts
  • S. Roy
    Departments of Medicine and Ophthalmology, Boston University School of Medicine, Boston, Massachusetts
  • Footnotes
    Commercial Relationships M. Wong, None; S. Roy, None.
  • Footnotes
    Support NIH
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 1356. doi:
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    • Get Citation

      M. Wong, S. Roy; Association Between High Glucose-Induced Overexpression of Collagen Type IV and Integrin Expression in Microvascular Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2007;48(13):1356.

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Abstract

Purpose:: To determine if downregulation of high glucose (HG)-induced collagen type IV (Col IV) overexpression is accompanied by a corresponding reduction in expression of integrin alpha-3, the subunit that specifically binds to Col IV.

Methods:: Rat microvascular endothelial cells were grown in normal (5 mM) or HG (30 mM) medium for 7 days and transfected with 0.4 uM Col IV antisense (AS-Col IV) oligonucleotides. Two days post-transfection, cells were harvested and protein analyzed by Western blot for Col IV and integrin alpha-3 expression. In parallel, cells plated on cover slips and similarly transfected with Col IV antisense oligonucleotides, were examined for Col IV and integrin alpha 3 immunoreactivity.

Results:: Using Western Blot analysis, a significant increase in Col IV expression and in integrin alpha-3 expression was observed in cells grown in HG medium compared to cells grown in N medium (133.0±17.0% of control, P<0.01) and (142.5±13.9% of control, P<0.01), respectively. In cells grown in HG medium and transfected with AS-Col IV oligonucleotides, Col IV expression and integrin alpha-3 expression was significantly reduced compared to cells grown in HG medium (101.6±20.7% vs 133.0±17.0% of control, P<0.04) and (116.5±9.1% vs 142.5±13.9% of control, P<0.03), respectively. Using Immunofluorescence staining, a significant increase in Col IV expression and in integrin alpha-3 expression was observed in cells grown in HG medium compared to cells grown in N medium (119.2±6.3% of control, P<0.01) and (116.9±11.4% of control, P<0.04), respectively. In cells grown in HG medium and transfected with AS-Col IV oligonucleotides, Col IV expression and integrin alpha-3 expression was significantly reduced compared to cells grown in HG medium (101.9±10.0% vs 119.2±6.3% of control, P<0.04) and (100.7±2.3% vs 116.9±11.4% of control, P<0.04), respectively.

Conclusions:: The findings indicate that high glucose-induced Col IV over expression can regulate corresponding integrin alpha-3 expression in rat microvascular endothelial cells. Col IV antisense oligonucleotide administration not only downregulates Col IV overexpression, but also has the added benefit of reducing integrin alpha-3 overexpression associated with retinal endothelial cells in the diabetic retina.

Keywords: diabetic retinopathy • diabetes 
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