RESUMO
PURPOSE: We identified the temporal expression of activator protein-1 (AP-1) and matrix metalloproteinases (MMPs) after linoleic acid hydroperoxide (LHP) induction of retinal neovascularization. METHODS: After injection of LHP into the vitreous of rabbits, samples were collected for AP-1 binding activity and mRNA for MMP-9 and MMPs activity. AP-1 binding activity was measured by electrophoretic mobility shift assay. MMP-9 activity was measured by zymography and mRNA by quantitative RT-PCR. RESULTS: AP-1 binding activity was increased at 1-3 hr. MMP-9 mRNA levels were increased at 3 hr in the neural retina and by 12 hr in the retinal pigment epithelium (RPE) layer. MMP-9 proteolytic activity was elevated within the neural retina and within the vitreous and in the RPE-interphotoreceptor matrix (IPM) at 12 hr and peaked at 24 hr or 4 days. CONCLUSIONS: LHP increases the transcription factor AP-1 which in turn may regulate retinal MMP-9 synthesis during neovascularization.
Assuntos
Ácidos Linoleicos/toxicidade , Peróxidos Lipídicos/toxicidade , Metaloproteinase 9 da Matriz/biossíntese , Retina/efeitos dos fármacos , Neovascularização Retiniana/induzido quimicamente , Fator de Transcrição AP-1/metabolismo , Animais , Cromatografia Líquida de Alta Pressão , Ensaio de Desvio de Mobilidade Eletroforética , Injeções , Masculino , Metaloproteinase 9 da Matriz/genética , RNA Mensageiro/metabolismo , Coelhos , Retina/metabolismo , Neovascularização Retiniana/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Transcrição AP-1/genética , Corpo VítreoRESUMO
PURPOSE: To study the effect of Sairei-to (ST), a Japanese traditional medicine, on oxygen-induced retinopathy (OIR) in rats. METHODS: OIR was induced by maintaining Sprague-Dawley neonatal rats in 80% oxygen for 12 days. The rats were treated once daily with oral administration of 0.75 g/kg (n = 9), 1.5 g/kg (n = 13) of ST in water, or water alone (WA, n = 13) at 5 mL/ kg body weight from day 6 to day 17. On day 18, retinal samples were collected. Retinal neovascularization (NV) was assessed by the NV score, and by the percentage of avascular area (% AVA), using a method previously reported. The number of severe retinal NV cases (NV > or = 9) was compared. The retinal vascular endothelial growth factor (VEGF) concentrations were measured with an immunoassay kit, at 0, 12, 24, 72 and 144 hours after oxygenation. RESULTS: NV score and % AVA decreased in the ST treated group compared to the WA group. However, severe NV was seen in five cases of WA and in one case of the ST treated group. Thus severe NV was inhibited significantly by ST treatment (p = 0.0185). Retinal VEGF did not differ between groups at any time points. CONCLUSION: These data suggest that severe NV in OIR is inhibited by ST treatment.
Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Neovascularização Retiniana/tratamento farmacológico , Animais , Animais Recém-Nascidos , Oxigênio/toxicidade , Ratos , Ratos Sprague-Dawley , Retina/química , Neovascularização Retiniana/etiologia , Fator A de Crescimento do Endotélio Vascular/análiseRESUMO
BACKGROUND: It is possible that oxidative stress causes several retinal diseases. However, the natural biogenic role of antioxidants in the retina is not clear. PURPOSE: This study investigates the change in concentration of vitamin E (VE), ascorbate and glutathione (GSH) in the retina following vitreous injection of 600 mug 18:2 linoleic acid hydroperoxide (LHP) in male New Zealand rabbits. METHOD: LHP was injected above the retinal surface. The animals were sacrificed and the eyes enucleated before LHP injection, 1, 3, 6, 12, 24 h and 4 and 7 days after LHP injection. Retinas were removed, VE and ascorbate measured by HPLC, and GSH determined by a fluorometric method. RESULTS: The concentration of VE in the retina decreased from pretreatment levels of 154.6 +/- 29.7 nmol/g wet weight (n = 7) and was lowest at 6 h (61.1 +/- 18.1 nmol/g wet weight, n = 4, p < 0.05), then increased gradually, returning slowly to pre-LHP levels by 7 days. The concentration of ascorbate in control retinas decreased at 6 h from pretreatment levels of 7.33 +/- 0.93 micromol/g wet weight (n = 7) to 2.74 +/- 0.16 micromol/g wet weight (n = 4, p < 0.05) and returned to pretreatment levels rapidly by 24 h after injection. The concentration of GSH in retinas decreased from baseline levels of 109.53 +/- 8.19 microg/g wet weight (n = 9), was lowest at 12 h (72.40 +/- 11.17 microg/g wet weight, n = 5, p < 0.05) and returned to pretreatment levels by 7 days. CONCLUSION: The results suggest that intravitreous LHP injection is a contributor to oxidative stress in the rabbit retina by causing a reduction in antioxidant capacity.