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WESTERN REGIONAL MEETING AT A GLANCE

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WAFMR, WSCI, WAP and WSPR Joint Plenary Session II 1:45 PM Thursday, January 31, 2008

1 EXPRESSION AND FUNCTION OF ERYTHROPOIETIN AND VASCULAR ENDOTHELIAL GROWTH FACTOR INCREASED IN OXYGEN INDUCED RETINOPATHY

S. Patel1,2, H. Chen2,3, N. London2,3, Z. Tong2,3, Z. Yang2,3, D. Li3, and K. Zhang2,3. University of Utah, Salt Lake City, UT; 2University of Utah, Salt Lake City, UT and 3University of Utah, Salt Lake City, UT.

Purpose of Study

Retinopathy of prematurity (ROP) is a prevalent morbidity among preterm infants and is characterized by retinal neovascularization. Recent research in diabetic retinopathy, thought to have similar pathogenesis to ROP, has shown that erythropoietin (Epo) and vascular endothelial growth factor (VEGF) are involved in neovascular angiogenesis. It is not known whether and how these growth factors are involved in ROP. We hypothesized that Epo and VEG-F mRNA expression is up-regulated in oxygen induced retinopathy (OIR) and these proteins are involved in endothelial cell proliferation, migration and permeability.

Methods Used

After exposure to hyperoxic conditioning, murine vitreous samples were obtained at postnatal day 17, retina isolated and RNA extracted for Epo and VEGF mRNA determination. This procedure was repeated in control mice. Migration and proliferation assays were performed according to manufacture's instruction (Promocell, Germany;Cambrex, Walkersville) on in vivo endothelial cells using increasing concentrations of Epo and VEGF. Cell growth and migration was measured manually and using Cell Counting Kit 8 (Dojindo Molecular Technologies, Gaithersburg). Finally, Epo was injected into the vitreous of murine retina, followed by intravenous injection of Evan's blue dye 6 hours later. Retinal vascular permeability was assessed by measuring Evan's blue leakage into vitreous cavity.

Summary of Results

Both Epo and VEGF mRNA levels were significantly greater in hyperoxia exposed mice (p < 0.001, n = 4). However, Epo levels were significantly higher than VEGF levels (15-fold increase vs. 2.5 fold increase, p < 0.05). In addition, both Epo and VEGF stimulated endothelial cell migration and proliferation in a dose dependent …

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