Effect of zeaxanthin and antioxidant supplementation on vascular endothelial growth factor (VEGF) expression in apolipoprotein-E deficient mice.
OBJECTIVE
Apolipoprotein E(-/-) deficient (apoE(-/-)) mice develop hypercholesterolemia, atherosclerosis, and retinal alterations. We studied the oxidative status and vascular endothelial growth factor (VEGF) expression in murine retinal pigment epithelium-choroid (RPE) and Bruch's membrane (BM) ultrastructure and the effect of zeaxanthin.


METHODS
Ten 6-month-old C57BL/6 and 40 apoE(-/-) mice were divided into four groups (n = 10 each) and fed different diets for 12 weeks based on body weight: wild type (WT) and apoE(-/-) (AE-Con) mice standard rodent chow; apoE(-/-) mice (AES) standard rodent chow with ascorbate (800 mg/kg), tocopherol (1053 mg/kg), and zinc (135 mg/kg); and apoE(-/-) mice the last diet plus zeaxanthin with either 0.4 g/kg (AES-Z04) or 4 g/kg feed (AES-Z4).


RESULTS
Plasma total cholesterol (TC) and triglycerides (TG) and urine lipid peroxidation (isoprostanes) were measured. VEGF expression was determined in RPE-choroid homogenates. Zeaxanthin uptake was assessed in liver and retina by high-performance liquid chromatography; the retinal ultrastructure was analyzed by electron microscopy. AE-Con mice had higher plasma TC (p < 0.001) and TG (p < 0.001) values than WT mice. AE-Con mice had higher RPE-choroid-VEGF levels than WT mice (p < 0.05), BM thickness (p < 0.001) and presence of basal laminar deposits (BLamD). AES-Z4 resulted in lower urinary isoprostanes (p = 0.054) and lower VEGF expression in the RPE-choroid (p < 0.01). BM in the AES-Z4 animals had less confluent BLamD than AE-Con, AES, or AES-Z04 animals.


CONCLUSIONS
We have reported that supplementation with zeaxanthin and antioxidants may delay or reverse alterations in the RPE and deposits in BM, and reduced VEGF expression observed in apoE(-/-) mice.
Fernández-Robredo
Patricia
P
Ophthalmology Research Laboratory, University of Navarra, Pamplona, Spain.
Recalde
Sergio
S
Arnáiz
Gonzalo
G
Salinas-Alamán
Angel
A
Sádaba
Luis M
LM
Moreno-Orduña
Maite
M
García-Layana
Alfredo
A
D000818:Animals; D000975:Antioxidants / Q000008:administration & dosage*; D001057:Apolipoproteins E / Q000172:deficiency*; D001205:Ascorbic Acid / Q000008:administration & dosage; D015153:Blotting, Western; D016570:Bruch Membrane / Q000378:metabolism / Q000648:ultrastructure; D002784:Cholesterol / Q000097:blood; D002829:Choroid / Q000378:metabolism* / Q000648:ultrastructure; D002851:Chromatography, High Pressure Liquid; D004032:Diet; D019587:Dietary Supplements; D015237:Dinoprost / Q000031:analogs & derivatives / Q000652:urine; D006952:Hyperlipoproteinemia Type III / Q000378:metabolism* / Q000473:pathology; D015227:Lipid Peroxidation; D008099:Liver; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D012160:Retina / Q000378:metabolism* / Q000648:ultrastructure; D055213:Retinal Pigment Epithelium / Q000378:metabolism / Q000648:ultrastructure; D024505:Tocopherols / Q000008:administration & dosage; D014280:Triglycerides / Q000097:blood; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*; D024341:Xanthophylls / Q000008:administration & dosage*; D065146:Zeaxanthins; D015032:Zinc / Q000008:administration & dosage
D000975:Antioxidants; D001057:Apolipoproteins E; D014280:Triglycerides; D042461:Vascular Endothelial Growth Factor A; D024341:Xanthophylls; D065146:Zeaxanthins; C467480:vascular endothelial growth factor A, mouse; C075750:8-epi-prostaglandin F2alpha; D002784:Cholesterol; D015237:Dinoprost; D015032:Zinc; D001205:Ascorbic Acid; D024505:Tocopherols