Procyanidins from grape seeds protect against phorbol ester-induced oxidative cellular and genotoxic damage.
OBJECTIVE
To evaluate the inhibitory effects of Vitis vinifera procyanidins (PAs) on carcinogen-induced oxidative stress.


METHODS
The single cell gel electrophoresis technique (comet assay) was employed to detect DNA damage induced by the carcinogen phorbol-12-myristate-13-acetate (PMA). The release of hydrogen peroxidase from polymorphonuclear leukocytes (PMNs) was assayed by the horseradish peroxidase-mediated oxidation of phenol red. The microplate assay was used to detect the presence of oxidative products by means of 2',7'-dichlorofluorescin-diacetate (DCFH-DA). The superoxide dismutase (SOD) activity of liver mitochondria was assayed, based on the ability of SOD to inhibit the generation of superoxidate anions by the xanthine-xanthine oxidase system. The malondialdehyde (MDA) level was determined by the thiobarbituric acid (TBA) assay.


RESULTS
DNA of NIH3T3 cells was significantly damaged after addition of PMA. The length of the comet tail was observed,while in normal cells the comet tail could not be observed. PAs showed significant protective effects on carcinogen PMA-induced DNA damage. Through assessment of DCFH-DA oxidation, PAs were shown to inhibit the PMA-induced release of hydrogen peroxide by PMNs, and to inhibit respiratory burst activity in NIH3T3 mouse fibroblasts. Ex vivo study showed that serum from rats administered with PAs displayed similar effects in a dose-dependent manner. In addition, PAs suppressed liver mitochondrial lipid peroxidation induced by PMA. PAs protected the activity of SOD and decreased the level of MDA in liver mitochondria damaged by PMA.


CONCLUSIONS
Dietary PAs from grape seeds protect against carcinogen-induced oxidative cellular and genotoxic damage.
Department of Pharmacology, Marine Drug Research Center, Nanjing University of Traditional Chinese Medicine, Nanjing 210029, China. luyingreen@163.net
Yin
Y
Lu
Wan-Zhou
WZ
Zhao
Zai
Z
Chang
Wen-Xin
WX
Chen
Lin
L
Li
D000818:Animals; D000975:Antioxidants / Q000302:isolation & purification / Q000494:pharmacology*; D044946:Biflavonoids / Q000302:isolation & purification / Q000494:pharmacology*; D002392:Catechin / Q000302:isolation & purification / Q000494:pharmacology*; D004249:DNA Damage / Q000187:drug effects*; D006861:Hydrogen Peroxide / Q000378:metabolism; D008315:Malondialdehyde / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008930:Mitochondria, Liver / Q000378:metabolism; D041681:NIH 3T3 Cells; D009504:Neutrophils / Q000378:metabolism*; D044945:Proanthocyanidins / Q000302:isolation & purification / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012639:Seeds / Q000737:chemistry; D013482:Superoxide Dismutase / Q000378:metabolism; D027843:Vitis / Q000737:chemistry
D000975:Antioxidants; D044946:Biflavonoids; D044945:Proanthocyanidins; D017382:Reactive Oxygen Species; C017674:procyanidin; D008315:Malondialdehyde; D002392:Catechin; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase