Modifying Choroidal Neovascularization Development with a Nutritional Supplement in Mice
Abstract
:1. Introduction
2. Materials and Methods
2.1. Study Animals
2.2. Diode Laser-Induced CNV Model
2.3. Oral Nutritional Supplements and the Vehicle
Per Capsule | Murine Dose Per Day | |
---|---|---|
Vitamin C (mg) | 120 | 1.57 |
Vitamin E (mg) | 15 | 0.20 |
Zinc (mg) | 7.5 | 0.10 |
Copper (µg) | 500 | 6.54 |
EPA (mg) | 190 | 2.48 |
DHA (mg) | 95 | 1.24 |
Lutein (mg) | 5 | 0.07 |
Zeaxanthin (mg) | 1 | 0.01 |
Resveratrol (mg) | 15 | 0.20 |
2.4. Intravitreal Treatments and the Injection Procedure
2.5. Evaluation of Leakage from Choroidal Neovascular Lesions
2.6. CD31 Immunofluorescence
2.7. Determination of Total Protein Levels
2.8. Vascular Endothelial Growth Factor and NOD-like Receptor Family Pyrin Domain Containing 3
2.9. Quantitative Real Time-Polymerase Chain Reaction
2.10. Matrix Metalloproteinase-2 and -9 Activity
2.11. Statistical Analyses
3. Results
3.1. Effect of Nutritional Supplements on Choroidal Neovascular Lesion Leakage
V | S | V + aVEGF | S + aVEGF | |
---|---|---|---|---|
FA | −0.030 ± 0.003 | −0.261 ± 0.04 *** | −0.112 ± 0.025 * | −0.091 ± 0.03 * |
CD31 (% vs. V) | 100 ± 10.03 | 63 ± 5.35 ** | 80 ± 9.62 * | 64 ± 9.69 * |
Protein expression | ||||
VEGF (% vs. V) | 100 ± 8.11 | 74.78 ± 11.96 * | 67.39 ± 10.30 * | 68.59 ± 9.70 * |
NLRP3 (% vs. V) | 100 ± 13.12 | 102.07 ± 4.26 | 108.08 ± 13.20 | 90.8 5 ± 12.83 |
Enzymatic activity | ||||
MMP2 (% vs. V) | 100 ± 6.49 | 80.45 ± 9.93 | 83.25 ± 6.96 | 104.30 ± 11.41 |
MMP9 (% vs. V) | 100 ± 10.08 | 105.50 ± 12.81 | 73.79 ± 10.64 | 54.75 ± 5.32 **, † |
Gene expression (Log RQ) | ||||
VEGF | 0.00 | −0.53 * | −0.48 * | −0.54 * |
MMP2 | 0.00 | −0.25 | −0.11 | −0.70 |
MMP9 | 0.00 | −0.18 | −0.09 | −0.29 *** |
3.2. Effect of Nutritional Supplements on Choroidal Neovascular Lesion Size
3.3. Effect of the Nutritional Supplement on Vascular Endothelial Growth Factor and NOD-Like Receptor Family Pyrin Domain Containing 3
3.4. Matrix Metalloproteinase Activity
3.5. Effect of the Nutritional Supplement on Gene Expression
4. Discussion
5. Conclusions
Supplementary Files
Supplementary File 1Acknowledgments
Author Contributions
Conflicts of Interest
References
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Ivanescu, A.A.; Fernández-Robredo, P.; Heras-Mulero, H.; Sádaba-Echarri, L.M.; García-García, L.; Fernández-García, V.; Moreno-Orduna, M.; Redondo-Exposito, A.; Recalde, S.; García-Layana, A. Modifying Choroidal Neovascularization Development with a Nutritional Supplement in Mice. Nutrients 2015, 7, 5423-5442. https://doi.org/10.3390/nu7075229
Ivanescu AA, Fernández-Robredo P, Heras-Mulero H, Sádaba-Echarri LM, García-García L, Fernández-García V, Moreno-Orduna M, Redondo-Exposito A, Recalde S, García-Layana A. Modifying Choroidal Neovascularization Development with a Nutritional Supplement in Mice. Nutrients. 2015; 7(7):5423-5442. https://doi.org/10.3390/nu7075229
Chicago/Turabian StyleIvanescu, Alina Adriana, Patricia Fernández-Robredo, Henar Heras-Mulero, Luis Manuel Sádaba-Echarri, Laura García-García, Vanessa Fernández-García, Maite Moreno-Orduna, Aitor Redondo-Exposito, Sergio Recalde, and Alfredo García-Layana. 2015. "Modifying Choroidal Neovascularization Development with a Nutritional Supplement in Mice" Nutrients 7, no. 7: 5423-5442. https://doi.org/10.3390/nu7075229