• Food Science and Biotechnology
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• v.17 no.2
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• pp.343-348
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• 2008

In this study, antioxidant activity of methanol extract of Glycyrrhiza uralensis Fisch (GUE) against $H_2O_2$-induced DNA damage in human leucocytcs and cell death in PC12 cells was determined. The effect of GUE on $H_2O_2$-induced DNA damage in human leucocytcs was evaluated by the comet assay, where GUE ($1-50\;{\mu}g/mL$) was a dose dependent inhibitor of DNA damage induced by $H_2O_2$. The protective effect of GUE against $H_2O_2$-induced damage on PC12 cells was investigated by MTT reduction assay and lactate dehydrogenase release assay. A marked reduction in cell survival induced by $H_2O_2$ was significantly prevented by $1-50\;{\mu}g/mL$ of GUE. The enzyme activity of caspase-3 was elevated in $H_2O_2$-treated PC12 cells, while preincubation with GUE for 30 min inhibited $H_2O_2$-induced caspase-3 activation in a dose-dependent manner. In conclusion, GUE ameliorates $H_2O_2$-induced DNA damage in human leucocytes and has neuroprotective effect by preventing cell death in PC12 cell, suggesting that GU may be a potential candidate for novel therapeutic agents for neuronal diseases associated with oxidative stress.

• Journal of Korean Medicine for Obesity Research
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• v.19 no.1
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• pp.1-11
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• 2019

Objectives: The purpose of the present study was to evaluate the preventive effects of ethanol extract of Polygalae radix (EEPR) against oxidative stress (hydrogen peroxide, $H_2O_2$)-induced DNA damage and apoptosis in Chang liver cells. Methods: Chang liver cells were pretreated with various concentrations of EEPR and then challenged with 0.5 mM $H_2O_2$. The cell viability and apoptosis were assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry analysis, respectively. The levels of reactive oxygen species (ROS), mitochondrial membrane potentials (MMPs) and adenosine tri-phosphate (ATP) contents were measured. Expression levels of Bcl-2 and Bax were also determined using Western blot analysis. Results: The results showed that the decreased survival rate induced by $H_2O_2$ could be attributed to the induction of DNA damage and apoptosis accompanied by the increased production of ROS, which was remarkably protected by EEPR. In addition, the loss of $H_2O_2$-induced MMPs and ATP contents was significantly attenuated in the presence of EEPR. The inhibitory effect of EEPR on $H_2O_2$-induced apoptosis was associated with up-regulation of Bcl-2 and down-regulation of Bax, thus reducing the Bax/Bcl-2 ratio. Conclusions: Our data prove that EEPR protects Chang liver cells against $H_2O_2$-induced DNA damage and apoptosis by scavenging ROS and thus suppressing the mitochondrial-dependent apoptosis pathway.

• Nutrition Research and Practice
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• v.4 no.3
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• pp.183-190
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• 2010

Blueberry was enzymatically hydrolyzed using selected commercial food grade carbohydrases (AMG, Celluclast, Termamyl, Ultraflo and Viscozyme) and proteases (Alcalase, Flavourzyme, Kojizyme, Neutrase and Protamex) to obtain water soluble compounds, and their protective effect was investigated against $H_2O_2$-induced damage in Chinese hamster lung fibroblast cell line (V79-4) via various published methods. Both AMG and Alcalase hydrolysates showed higher total phenolic content as well as higher cell viability and ROS scavenging activities, and hence, selected for further antioxidant assays. Both AMG and Alcalase hydrolysates also showed higher protective effects against lipid peroxidation, DNA damage and apoptotic body formation in a dose-dependent fashion. Thus, the results indicated that water soluble compounds obtained by enzymatic hydrolysis of blueberry possess good antioxidant activity against $H_2O_2$-induced cell damage in vitro.

• Nutrition Research and Practice
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• v.8 no.2
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• pp.138-145
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• 2014

BACKGROUND/OBJECTIVES: This study was performed to investigate the in vitro antioxidant and cytoprotective effects of fermented sesame sauce (FSeS) against hydrogen peroxide ($H_2O_2$)-induced oxidative damage in renal proximal tubule LLC-PK1 cells. MATERIALS/METHODS: 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical ($^{\bullet}OH$), and $H_2O_2$ scavenging assay was used to evaluate the in vitro antioxidant activity of FSeS. To investigate the cytoprotective effect of FSeS against $H_2O_2$-induced oxidative damage in LLC-PK1 cells, the cellular levels of reactive oxygen species (ROS), lipid peroxidation, and endogenous antioxidant enzymes including catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-px) were measured. RESULTS: The ability of FSeS to scavenge DPPH, $^{\bullet}OH$ and $H_2O_2$ was greater than that of FSS and AHSS. FSeS also significantly inhibited $H_2O_2$-induced ($500{\mu}M$) oxidative damage in the LLC-PK1 cells compared to FSS and AHSS (P < 0.05). Following treatment with $100{\mu}g/mL$ of FSeS and FSS to prevent $H_2O_2$-induced oxidation, cell viability increased from 56.7% (control) to 83.7% and 75.6%, respectively. However, AHSS was not able to reduce $H_2O_2$-induced cell damage (viability of the AHSS-treated cells was 54.6%). FSeS more effectively suppressed $H_2O_2$-induced ROS generation and lipid peroxidation compared to FSS and AHSS (P < 0.05). Compared to the other sauces, FSeS also significantly increased cellular CAT, SOD, and GSH-px activities and mRNA expression (P < 0.05). CONCULUSIONS: These results from the present study suggest that FSeS is an effective radical scavenger and protects against $H_2O_2$-induced oxidative damage in LLC-PK1 cells by reducing ROS levels, inhibiting lipid peroxidation, and stimulating antioxidant enzyme activity.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.5
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• pp.520-526
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• 2017

Carnosine was recently reported to protect against the DNA damage induced by oxidative stress. In this study, we investigated the protective effect of eel Anguilla japonica carnosine extracts prepared using different methods (heat treatment extracts, HTEs; ion exchange chromatography, IEC; ultrafiltration permeation, UFP) on leukocyte DNA damage using the comet assay. Human leukocytes were incubated with extracts of eel carnosine at concentrations (of 10, 50, $100{\mu}g/mL$), and then subjected to an oxidative stimulus [$200{\mu}M$ hydrogen peroxide ($H_2O_2$)]. Pretreatment of the cells for 30 min with carnosine significantly reduced the genotoxicity of $H_2O_2$ measured as DNA strand breaks. The protective effects of the three types of extract (HTE, IEC, and UFP) increased with concentration. At the highest concentration (100 g/mL). there were no statistical differences in oxidative damage between each extract treatment and PBS-treated negative controls. When leukocytes were incubated with carnosine for 30 min after exposure to $H_2O_2$. the protective ability of each extract changed. Therefore, eel carnosine inhibits the $H_2O_2$ induced damage to cellular DNA in human leukocytes, supporting the protective effect of this compound against oxidative damage.

• Journal of The Korean Society of Integrative Medicine
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• v.11 no.2
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• pp.169-179
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• 2023

Purpose : Although human periodontal ligament stem cells (hPDLSCs) are a supportive factor for tissue engineering, oxidative stress during cell culture and transplantation has been shown to affect stem cell viability and mortality, leading to failed regeneration. The aim of this study was to evaluate the antioxidant and protective effects against cell damage of celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, and the antioxidant signal of hPDLSCs in H₂O₂-induced oxidative stress. Methods : To induce oxidative stress in cultured hPDLSCs, H₂O₂ was used as an exogenous reactive oxygen species (ROS). Dose-dependent celecoxib (.1, 1, 10, or 100 µM) was administered after H₂O₂ treatment. WST-1 assay was used to assess cell damage and western blot was used to observe antioxidant activity of hPDLSCs in oxidative stress. Immunohistochemistry was performed for inverting the localization of the SOD and Nrf2 antibody. Results : We found that progressive cell death was induced in hPDLSCs by H₂O₂ treatment. However, low-dose celecoxib reduced H₂O₂-induced cellular damage and eventually enhanced the SOD activity and Nrf2 signal of hPDLSCs. Oxidative stress-induced morphological change in hPDLSCs included lowered the survival and number of spindle-shaped cells, and shrinkage and shortening of cell fibers. Notably, celecoxib promoted cell survival function and activated antioxidants such as SOD and Nrf2 by positively regulating the cell survival signal pathway, and also reduced the number of morphological changes in hPDLS. Immunohistochemistry results showed a greater number of SOD- and Nrf2-stained cells in the celecoxib-treated group following oxidative stress. Conclusion : By increasing SOD and Nrf2 expression at the antioxidant system, the findings suggest that celecoxib enhanced the antioxidative ability of hPDLSCs and protected cell viability against H₂O₂-induced oxidative stress by increasing SOD and Nrf2 expression in the antioxidant system.

• BMB Reports
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• v.43 no.10
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• pp.683-687
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• 2010

Previous studies have shown that one of the primary causes of increased iron content in the brain may be the release of excess iron from intracellular iron storage molecules such as ferritin. Free iron generates ROS that cause oxidative cell damage. Carnosine and related compounds such as endogenous histidine dipetides have antioxidant activities. We have investigated the protective effects of carnosine and homocarnosine against oxidative damage of DNA induced by reaction of ferritin with $H_2O_2$. The results show that carnosine and homocarnosine prevented ferritin/$H_2O_2$-mediated DNA strand breakage. These compounds effectively inhibited ferritin/$H_2O_2$-mediated hydroxyl radical generation and decreased the mutagenicity of DNA induced by the ferritin/$H_2O_2$ reaction. Our results suggest that carnosine and related compounds might have antioxidant effects on DNA under pathophysiological conditions leading to degenerative damage such as neurodegenerative disorders.

• Archives of Pharmacal Research
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• v.29 no.2
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• pp.159-164
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• 2006

The protective properties of Satureja hortensis L. on the rat lymphocytes DNA lesions were tested. Lymphocytes were isolated from blood samples taken from healthy rats. DNA breaks and resistance to $H_{2}O_{2}$-induced damage were measured with the comet assay. Rat lymphocytes were incubated in S. hortensis ethanolic extract (SHE) (0.05, 0.1, 0.5, 1.0, and 2.5 mg/mL), essential oil (SHEO)(0.05, 0.1, 0.5, 1.0, and 2.5 ${mu}L/mL$), $H_{2}O_{2}$ (50, 100, and 200 ${\mu}M$), a combination of $H_{2}O_{2}$ (200 mM) with either SHE (1.0, 2.5 mg/mL) or SHEO (1.0, 2.5 ${\mu}L/mL$) at $4^{\circ}C$ for 30 min, and the extent of DNA migration was measured using a single-cell microgel electrophoresis technique under alkaline conditions. Treatment of rat lymphocytes with SHE or SHEO resulted in significant reduction of $H_{2}O_{2}$-induced DNA damage compared to controls. SHE exhibited a significant (P<0.01) inhibitory effect on oxidative DNA damage at 2.5 mg/mL. SHEO (1.0 and 2.5 ${\mu}L/mL$) also showed significant inhibitory effects (P<0.01) on $H_{2}O_{2}$ induced chromosomal damage. In conclusion both the ethanolic extract and the essential oil of the plant reversed the oxidative damage to rat lymphocytes induced by hydrogen peroxide.

• Journal of Acupuncture Research
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• v.18 no.1
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• pp.88-99
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• 2001

Objectives : This study was performed to determine if Orostachys japonicus A. Berger aquacupuncture (OjB) provides the protective effect against the loss of celi viability and DNA damage induced by oxidant in renal proximal tubular cells. Methods : The cell viability was evaluated by a MTT reduction assay and DNA damage was estimated by measuring double stranded DNA breaks in opossum kidney (OK) cells, an established proximal tubular cell line. Lipid peroxidation was determined by measuring malondialdehyde (MDA), a product of lipid peroxidation. Results : $H_2O_2$ increased the loss of cell viability in a time-dependent manner, which were prevented by 0.1% OjB. The protective effect of OjB was dose-dependent over concentration range of 0.05-0.5%. $H_2O_2$ caused ATP depletion and DNA damage, which were prevented by OjB and the hydrogen peroxide scavenger catalase. The loss of cell viability by $H_2O_2$ was not affected by the antioxidant DPPD, but lipid peroxidation by the oxidant was completely inhibited by DPPD. Conclusions : These data suggest that $H_2O_2$-induced death results from a lipid peroxidation-independent mechanism and the protective effect of OjB is not associated with its antioxidant activity.

• Herbal Formula Science
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• v.25 no.2
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• pp.179-191
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• 2017

Objectives : Increased oxidative stress by reactive oxygen species (ROS) has been suggested as a major cause of muscle fatigue. Although several studies have demonstrated the various biological properties of Sophora flavescens Aiton, Glycyrrhiza uralensis Fischer and Dictamnus dasycarpus Turcz, but the antioxidative potentials have not been clearly demonstrated. The present study was designed to investigate the protective effects of their water and ethanol extract mixtures (medicinal herbal mixtures, MHMIXs) on hydrogen peroxide ($H_2O_2$)-induced cell damage and apoptosis in C2C12 myoblasts. Methods : Cytotoxicity was assessed by an MTT assay. Quantitative evaluation of apoptosis induction and ROS production was evaluated by flow cytometry analysis. Expression levels of apoptosis regulatory and DNA-damage proteins were detected by Western blotting. Result : The inhibition of $H_2O_2$-induced cell proliferation was effectively blocked in extracts of 3: 1: 1 (EMHMIXs-1) or 2: 2: 1 (EMHMIXs-2) of S. flavescens, G. uralensis and D. dasycarpus Turcz, ethanol extracts from various complex extracts in C2C12 myoblasts. EMHMIXs-1 and EMHMIXs-2 also effectively attenuated $H_2O_2$-induced C2C12 cell apoptosis, which was associated with the restoration of the upregulation of Bad and death receptor 4, and downregulation of XIAP and cIAP-1 induced by $H_2O_2$. In addition, these herbal mixtures significantly blocked the $H_2O_2$-induced ROS generation and phosphorylation of $p-{\gamma}H2A.X$, which suggests that they can prevent $H_2O_2$-induced cellular DNA damage. Conclusions : The results suggest that EMHMIXs-1 and EMHMIXs-2 could block the DAN damage and apoptosis of C2C12 myoblasts by oxidative stress through blocking ROS generation.