Effect of grape polyphenols on oxidative stress in canine lens epithelial cells

Curtis A. Barden Department of Veterinary Clinical Sciences, College of Veterinary Medicine, and the Department of Human Nutrition, College of Education and Human Ecology (Bomser), The Ohio State University, Columbus, OH 43210.

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Heather L. Chandler Department of Veterinary Biosciences, College of Veterinary Medicine, and the Department of Human Nutrition, College of Education and Human Ecology (Bomser), The Ohio State University, Columbus, OH 43210.

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Ping Lu Department of Veterinary Clinical Sciences, College of Veterinary Medicine, and the Department of Human Nutrition, College of Education and Human Ecology (Bomser), The Ohio State University, Columbus, OH 43210.

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Joshua A. Bomser Department of Veterinary Clinical Sciences, College of Veterinary Medicine, and the Department of Human Nutrition, College of Education and Human Ecology (Bomser), The Ohio State University, Columbus, OH 43210.
Department of Veterinary Biosciences, College of Veterinary Medicine, and the Department of Human Nutrition, College of Education and Human Ecology (Bomser), The Ohio State University, Columbus, OH 43210.
Department of Veterinary Clinical Sciences, College of Veterinary Medicine, and the Department of Human Nutrition, College of Education and Human Ecology (Bomser), The Ohio State University, Columbus, OH 43210.
Department of Veterinary Clinical Sciences, College of Veterinary Medicine, and the Department of Human Nutrition, College of Education and Human Ecology (Bomser), The Ohio State University, Columbus, OH 43210.

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Carmen M. H. Colitz Department of Veterinary Clinical Sciences, College of Veterinary Medicine, and the Department of Human Nutrition, College of Education and Human Ecology (Bomser), The Ohio State University, Columbus, OH 43210.

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 DVM, PhD

Abstract

Objective—To evaluate whether the effects of oxidative stress could be attenuated in cultures of canine lens epithelial cells (LECs) by incubation with grape seed proanthocyanidin extract (GSE), resveratrol (RES), or a combination of both (GSE+RES).

Sample Population—Primary cultures of canine LECs.

Procedures—LECs were exposed to 100MM tertiary butyl-hydroperoxide (TBHP) with or without GSE, RES, or GSE+RES. The dichlorofluorescein assay was used to detect production of reactive oxygen species (ROS), and immunoblot analysis was used to evaluate the expression of stress-induced cell-signaling markers (ie, the mitogen-activated protein kinase [MAPK] and phosphoinositide-3 kinase [PI3K] pathways).

Results—GSE and GSE+RES significantly reduced ROS production after a 30-minute exposure to TBHP. Only GSE significantly reduced ROS production after a 120-minute exposure to TBHP. Incubation with GSE reduced TBHP-induced activity of the MAPK and PI3K pathways.

Conclusions and Clinical Relevance—GSE inhibited key components associated with cataractogenesis, ROS production, and stress-induced cell signaling. On the basis of the data reported here, there is strong evidence that GSE could potentially protect LECs from the damaging effects of oxidative stress.

Abstract

Objective—To evaluate whether the effects of oxidative stress could be attenuated in cultures of canine lens epithelial cells (LECs) by incubation with grape seed proanthocyanidin extract (GSE), resveratrol (RES), or a combination of both (GSE+RES).

Sample Population—Primary cultures of canine LECs.

Procedures—LECs were exposed to 100MM tertiary butyl-hydroperoxide (TBHP) with or without GSE, RES, or GSE+RES. The dichlorofluorescein assay was used to detect production of reactive oxygen species (ROS), and immunoblot analysis was used to evaluate the expression of stress-induced cell-signaling markers (ie, the mitogen-activated protein kinase [MAPK] and phosphoinositide-3 kinase [PI3K] pathways).

Results—GSE and GSE+RES significantly reduced ROS production after a 30-minute exposure to TBHP. Only GSE significantly reduced ROS production after a 120-minute exposure to TBHP. Incubation with GSE reduced TBHP-induced activity of the MAPK and PI3K pathways.

Conclusions and Clinical Relevance—GSE inhibited key components associated with cataractogenesis, ROS production, and stress-induced cell signaling. On the basis of the data reported here, there is strong evidence that GSE could potentially protect LECs from the damaging effects of oxidative stress.

Contributor Notes

Supported by IAMS Canine Research Funds.

Presented at the 37th Annual Conference of the American College of Veterinary Ophthalmologists, San Antonio, Tex, October–November 2006, and the Annual Meetings of the Association for Research in Vision and Ophthalmology, Fort Lauderdale, Fla, May 2006 and May 2007.

The authors thank Drs. Donna Kusewitt and Anne Metzler for technical assistance.

Address correspondence to Dr. Colitz.
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