Abstract
Methyl gallate (meGAL) is known as one of major antioxidants. To investigate whether meGAL protects human cells from oxidative stress, meGAL extracted from Korean medicinal plant, Cercis chinensis leaves, was primarily screened using cell viability assay against oxidative stress. Human umbilical vein endothelial cells (HUVECs) were treated with three different concentrations of meGAL for indicated time. After or during meGAL treatment, H(2)O(2) was added and incubated. meGAL showed free radical scavenging effect at low concentration (0.02 mM) and cell protective effect against H2O2-mediated oxidative stress. meGAL recovered viability of HUVECs damaged by H(2)O(2)-treatment, reduced the lipid peroxidation (LPO) and decreased the internal reactive oxygen species (ROS) level elevated by H(2)O(2)-treatment. Free radical scavenging effect of meGAL was proven to be very high. Differential display reverse transcription-PCR analysis showed that meGAL upregulated the levels of regulator of chromatin condensation 1, type 1 sigma receptor and phosphate carrier protein expressions, respectively. Based on structural similarity compared with meGAL, 14 chemicals were chosen and viability assay was performed. Four chemicals, haematommic acid (56.2% enhancement of viability), gallic acid (35.0%), methylorsellinic acid (23.7%), and syringic acid (20.8%), enhanced more potent cell viability than meGAL, which showed only 18.1% enhancement of cell viability. These results suggest that meGAL and four meGAL-related chemicals protect HUVECs from oxidative stress.
Highlights
reactive oxygen species (ROS) and reactive nitrogen species (RNS) constantly generated in normal condition by aerobic metabolism include free radicals such as superoxide anion, hydroxyl radicals, nonradical hydrogen peroxide, peroxinitrite, nitroxyl anion and nitric oxide (Beckman and Ames, 1998; Curtin et al, 2002; Droge, 2002)
Cell based assay is basically to monitor the change of cell viability induced by oxidative stress, which was H2O2 exposure on Human umbilical vein endothelial cells (HUVECs) in our experiments
Many natural products including flavonoids, cumarins, polyols have been studied for the characterization and the development as antioxidative reagents (Finkel and Holbrook, 2000; Lee et al, 2002)
Summary
ROS and reactive nitrogen species (RNS) constantly generated in normal condition by aerobic metabolism include free radicals such as superoxide anion, hydroxyl radicals, nonradical hydrogen peroxide, peroxinitrite, nitroxyl anion and nitric oxide (Beckman and Ames, 1998; Curtin et al, 2002; Droge, 2002). Redox homeostasis is maintained by controlling the balance between ROS production and various types of scavengers called antioxidants. Transient changes in oxidants-antioxidant balance are normally regulated by changing the production of counter species and reached to the steady-state over time (Shuli et al, 1991; Nakamura et al, 1994; Adler et al, 1999; Zhang and Storz, 2000; Droge, 2002). The persistent production of abnormally large amount of ROS or RNS, 344 Exp. Mol. Vol 37(4), 343-352, 2005 may lead to persistent changes in signal transduction and gene expression, which in turn may give rise to certain diseases
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