Grape seed proanthocyanidin extract protects human lens epithelial cells from oxidative stress via reducing NF-кB and MAPK protein expression.
OBJECTIVE
Oxidative damage induced by H₂O₂ treatment can irreversibly damage the lens epithelium, resulting in cell death and cataract. Grape seed extract (GSE) is a widely consumed dietary supplement that has the capability to scavenge oxidants and free radicals. GSE contain 70%-95% standardized proanthocyanidins. The study described herein investigated the protective effect of Grape seed proanthocyanidin extract (GSPE) on H₂O₂-induced oxidative stress in human lens epithelial B-3 (HLEB-3) cells and the possible molecular mechanism involved.


METHODS
HLE-B3 cells exposed to different doses of H₂O₂ were cultured with various concentrations of GSPE and subsequently monitored for cell viability by the 4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT) assay. The apoptosis rate and ROS generation were detected by flow cytometric analysis. Expression of NF-кB/P65 and mitogen activated protein kinase (MAPK) proteins were measured by western blot.


RESULTS
GSPE clearly reduced H₂O₂ induced cell apoptosis and reactive oxygen species (ROS) generation and protected HLEB-3 cells from H₂O₂ induced oxidative damage. GSPE depressed H₂O₂-induced activation and translocation of NF-кB/p65. GSPE also depressed H₂O₂-induced phosphorylation of the p38 and c-Jun N-terminal kinase (JNK) proteins of the MAPK family at various time points studied.


CONCLUSIONS
GSPE could be useful in attenuation of H₂O₂-induced oxidative stress and the activation of NF-кB and MAPK signaling in HLE-B3 cells, which suggests that GSPE has a potential protective effect against cataractogenesis.
Department of Ophthalmology, The First Affiliated Hospital of Harbin Medical University, Harbin, P.R. China.
Zhiyan
Z
Jia
Zhen
Z
Song
Yuhui
Y
Zhao
Xiurong
X
Wang
Ping
P
Liu
D002460:Cell Line; D004847:Epithelial Cells / Q000187:drug effects*; D005434:Flow Cytometry / Q000379:methods; D005609:Free Radicals; D015971:Gene Expression Regulation, Enzymologic; D056604:Grape Seed Extract / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D007908:Lens, Crystalline / Q000187:drug effects*; D020935:MAP Kinase Signaling System; D016328:NF-kappa B / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D010766:Phosphorylation; D044945:Proanthocyanidins / Q000494:pharmacology*; D017382:Reactive Oxygen Species
D005609:Free Radicals; D056604:Grape Seed Extract; D016328:NF-kappa B; D044945:Proanthocyanidins; D017382:Reactive Oxygen Species; C013221:proanthocyanidin; D006861:Hydrogen Peroxide; D048031:JNK Mitogen-Activated Protein Kinases
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