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Natural Product Research Part A
Year: 2010  |  Volume: 24  |  Issue: 7  |  Page No.: 599 - 609

Effect of fresh apple extract on glycated protein/iron chelate-induced toxicity in human umbilical vein endothelial cells in vitro

Ikuo Nishigaki, Balasubramanian Rajkapoor, Peramaiyan Rajendran, Ramachandran Venugopal, Ganapathy Ekambaram, Dhanapal Sakthisekaran and Yutaka Nishigaki    

Abstract: Consumption of fruits and vegetables has been associated with a low incidence of cardiovascular and other chronic diseases. The present study was aimed at evaluating the protective effects of fresh apple extract (AE) on human umbilical vein endothelial cells (HUVEC) exposed to cytotoxic glycated protein (GFBS)/iron (FeCl3) chelate. The experimental design comprised 10 groups with 5 flasks in each group. Group I was treated with 15% foetal bovine serum (FBS). Groups II, III and IV were treated with GFBS (70 µM), FBS + FeCl3 (20 µM), and GFBS + FeCl3, respectively. The other six groups were as follows: Group V, GFBS + AE (100 µg); Group VI, FBS + FeCl3 + AE (100 µg); Group VII, GFBS + FeCl3 + AE (100 µg); Group VIII, GFBS + AE (250 µg); Group IX, FBS + FeCl3 + AE (250 µg); and Group X, GFBS + FeCl3 + AE (250 µg). After 24 h incubation, cells were collected from all the experimental groups and assessed for lipid peroxidation (LPO) and activities of the antioxidant enzymes cytochrome c reductase and glutathione S-transferase (GST). HUVEC incubated with glycated protein (GFBS) either alone or combined with iron chelate showed a significant (p < 0.001) elevation of LPO accompanied by depletion of superoxide dismutase, catalase, glutathione peroxidase (GPx) and glutathione reductase (GR), in addition to increased microsomal cytochrome c reductase and decreased GST activities. Treatment of GFBS- or GFBS + FeCl3-exposed HUVEC with AE at 100 or 250 µg significantly decreased the level of LPO and returned the levels of antioxidants cytochrome c reductase and GST to near normal in a dose-dependent manner. The extracts recovered viability of HUVEC damaged by GFBS-iron treatment in a concentration-dependent manner. These findings suggest a protective effect of AE on HUVEC against glycated protein/iron chelate-induced toxicity, which suggests that AE could exert a beneficial effect by preventing diabetic angiopathies.

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