• Journal of Pharmacopuncture
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• v.19 no.1
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• pp.59-69
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• 2016

Objectives: The mushroom Ganoderma lucidum has been widely used as a traditional herbal medicine for many years. Although several studies have focused on the anti-oxidative activity of this mushroom, the molecular mechanisms underlying its activity have not yet been clearly established. The present study investigated the cytoprotective effect of ethanol extract of Ganoderma lucidum (EGL) against oxidative stress (hydrogen peroxide, $H_2O_2$) and elucidated the underlying mechanisms in a C2C12 myoblast cell line. Methods: Oxidative stress markers were determined by using the comet assay to measure reactive oxygen species (ROS) generation and deoxyribonucleic acid (DNA) damage. Cell viability and Western blotting analyses were employed to evaluate the cellular response to EGL and $H_2O_2$ in C2C12 cells. Transfection with nuclear factor erythroid 2-related factor 2 (Nrf2)-specific small interfering ribonucleic acid (siRNA) was conducted to understand the relationship between Nrf2 expression and $H_2O_2$-induced growth inhibition. Results: The results showed that EGL effectively inhibited $H_2O_2$-induced growth and the generation of ROS. EGL markedly suppressed $H_2O_2$-induced comet-like DNA formation and phosphorylation of histone H2AX at serine 139 ($p-{\gamma}H2AX$), a widely used marker of DNA damage, suggesting that EGL prevented $H_2O_2$-induced DNA damage. Furthermore, the EGL treatment effectively induced the expression of Nrf2, as well as heme oxygenase-1 (HO-1), with parallel phosphorylation and nuclear translocation of Nrf2 in the C2C12 myoblasts. However, zinc protoporphyrin IX, a HO-1 inhibitor, significantly abolished the protective effects of EGL against $H_2O_2$-induced accumulation of ROS and reduced cell growth. Notably, transient transfection with Nrf2-specific siRNA attenuated the cytoprotective effects and HO-1 induction by EGL, indicating that EGL induced the expression of HO-1 in an Nrf2-dependent manner. Conclusion: Collectively, these results demonstrate that EGL augments the cellular anti-oxidant defense capacity through activation of Nrf2/HO-1, thereby protecting C2C12 myoblasts from $H_2O_2$-induced oxidative cytotoxicity.

• Korean Journal of Veterinary Research
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• v.44 no.3
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• pp.475-482
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• 2004

For the induction of arthropathy, 4% hydrogen peroxide ($H_2O_2$) was injected for 4 weeks into the intra-articular space of the 25 New Zealand white rabbits to damage articular cartilage. The verification of arthropathy induction and the effect of scan-bio laser treatment were determined by measuring superoxide dismutase (SOD) activity, by observing gross and histopathologic findings. The SOD activity increased by about 40% in arthropathy group, as compared to controls. Although SOD activity in arthropathy group was not significantly different from the 2-week group, it was significantly different from the 4-week control and treatment groups. There was also a significant difference between the 4-week control and treatment groups. Grossly, erosions formed on the articular cartilage surface, and the lateral femoral condyle was damaged in arthropathy group. In comparison, there was slight, but not significant, progression of the lesion in the 2-week control group, and no difference between the 2-week treatment and control groups. Conversely, severe erosions damaged the articular cartilage in the 4-week control group. Cartilage proliferation was seen in gross observations in the 4-week treatment group, suggesting a treatment effect. Histopathologically, there was slight articular surface damage and apoptosis in arthropathy group, and serious cartilage damage, despite slight chondrocyte proliferation, in the 4-week control group. By contrast, the 4-week treatment group showed chondrocyte replacement, with close to normal articular cartilage on the articular surface. There was significant cartilage proliferation with regeneration of the articular cartilage on the articular surface in the group treated with low-level laser, as compared to control group, when arthropathy was induced by $H_2O_2$ injections. Therefore, low-level laser was effective in the treatment of chemically induced arthropathy.

• Korean Journal of Acupuncture
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• v.24 no.1
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• pp.171-187
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• 2007

Objectives : This study was performed to determine if Orostachys japonicus A. Berger herbal acupuncture (OjB) provides the protective effect against the loss of cell 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 : H2O2 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%. H2O2 caused ATP depletion and DNA damage, which were prevented by OjB and the hydrogen peroxide scavenger catalase. The loss of cell viability by H2O2 was not affected by the antioxidant DPPD, but lipid peroxidation by the oxidant was completely inhibited by DPPD. Generation of superoxide and H2O2 in neutrophils activated by phorbol-12,13-dibutyrate was inhibited by OjB in a dose-dependent manner. OjB inhibited generation of H2O2 in OK cells treated with antimycin A and exerted a direct H2O2 scavenging effect. Exposure of OK cells to 1 mM tBHP caused a significant depletion of glutathione which was prevented by OjB. OjB accelerated the recovery in cells cultured for 20 hr in normal medium without oxidant following oxidative stress. Conclusions : These results suggest that OjB exerts the protective effect against oxidant-induced cell injury and its protective effect was resulted from radical scavenging and antioxidant activities.

• Biomolecules & Therapeutics
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• v.32 no.3
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• pp.329-340
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• 2024

Mangiferin is a kind of natural xanthone glycosides and is known to have various pharmacological activities. However, since the beneficial efficacy of this compound has not been reported in retinal pigment epithelial (RPE) cells, this study aimed to evaluate whether mangiferin could protect human RPE ARPE-19 cells from oxidative injury mimicked by hydrogen peroxide (H₂O₂). The results showed that mangiferin attenuated H₂O₂-induced cell viability reduction and DNA damage, while inhibiting reactive oxygen species (ROS) production and preserving diminished glutathione (GSH). Mangiferin also antagonized H₂O₂-induced inhibition of the expression and activity of antioxidant enzymes such as manganese superoxide dismutase and GSH peroxidase, which was associated with inhibition of mitochondrial ROS production. In addition, mangiferin protected ARPE-19 cells from H₂O₂-induced apoptosis by increasing the Bcl-2/Bax ratio, decreasing caspase-3 activation, and blocking poly(ADP-ribose) polymerase cleavage. Moreover, mangiferin suppressed the release of cytochrome c into the cytosol, which was achieved by interfering with mitochondrial membrane disruption. Furthermore, mangiferin increased the expression and activity of heme oxygenase-1 (HO-1) and nuclear factor-erythroid-2 related factor 2 (Nrf2). However, the inhibition of ROS production, cytoprotective and anti-apoptotic effects of mangiferin were significantly attenuated by the HO-1 inhibitor, indicating that mangiferin promoted Nrf2-mediated HO-1 activity to prevent ARPE-19 cells from oxidative injury. The results of this study suggest that mangiferin, as an Nrf2 activator, has potent ROS scavenging activity and may have the potential to protect oxidative stress-mediated ocular diseases.

• Journal of Life Science
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• v.31 no.2
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• pp.126-136
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• 2021

Oxidative stress causes injury to and degeneration of retinal pigment epithelial (RPE) cells. It is involved in several retinal disorders and leads to vision loss. In the present study, we investigated the effect of 14 kinds of natural compounds and two kinds of synthetic compounds on oxidative stress-induced cellular damage in human PRE cell lines (ARPE-19). From among them, we selected five kinds of compounds, including auranofin, FK-509, hemistepsin A, honokiol, and spermidine, which have inhibitory effects against hydrogen peroxide (H2O2)-mediated cytotoxicity. In addition, we found that four kinds of compounds (excluding auranofin) have protective effects on H2O2-induced mitochondrial dysfunction. Furthermore, the expression of phosphorylation of histone H2AX, a sensitive marker of DNA damage, was markedly up-regulated by H2O2, whereas it was notably down-regulated by FK-506, honokiol, and spermidine treatment. Meanwhile, five kinds of candidate compounds had no effect on H2O2-induced intracellular reactive oxygen species (ROS) levels, suggesting that the five candidate compounds have protective effects on oxidative stress-induced cellular damage through the ROS-independent pathway. Taken together, according to the results of H2O2-mediated cellular damage―such as cytotoxicity, apoptosis, mitochondrial dysfunction, and DNA damage―spermidine and FK-506 are the natural and synthetic compounds with the most protective effects against oxidative stress in RPE. Although further studies on the identification of the mechanism responsible are required, the results of the present study suggest the possibility of using spermidine and FK-506 to suppress the risk of retinal disorders.

• Journal of Life Science
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• v.28 no.6
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• pp.708-717
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• 2018

The antioxidant activity and protective effects of a hot water extract from the Stauntonia hexaphylla fruit (WESHF) were investigated in vitro and in vivo. The total polyphenol and flavonoid contents of WESHF were $16.13{\pm}0.27mg$ gallic acid equivalent/g and $4.7{\pm}0.80mg$ catechin equivalent/g, respectively. In addition, the DPPH radical-scavenging activity ($SC_{50}$) and the Oxygen Radical Absorbance capacity of WESHF were $63.62{\pm}4.10{\mu}g/ml$ and $90.63{\pm}5.29{\mu}M$ trolox equivalent/g, respectively. The hepatoprotective effect of WESHF against hydrogen peroxide-induced oxidative damage was investigated. $H_2O_2$-induced liver damage on HepG2 cells was prevented by $200{\mu}g/ml$ of WESHF. Furthermore, to investigate the protection mechanism of WESHF on hydrogen peroxide-induced cytotoxicity in HepG2 cells, pre-treatment with $200{\mu}g/ml$ of WESHF significantly attenuated a decrease in the activities of CAT, SOD, GR, and GPx. The hepatoprotective activity of WESHF was evaluated in an experimental model of hepatic damage induced by acetaminophen (APAP). The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were significantly decreased in the livers of mice treated with 200 mg/kg of WESHF compared to the APAP-treated group. The lipid peroxidation level, which increased after APAP administration, was significantly reduced in the WESHF group. In addition, histological examinations of the liver showed the same protective effect of WESHF treatment. Based on these findings, it is suggested that WESHF has potent hepatoprotective effects, and the mechanism that causes this type of protection could be related to antioxidant pathways.

• Journal of Life Science
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• v.27 no.9
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• pp.1070-1077
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• 2017

Chondrocyte apoptosis induced by reactive oxygen species (ROS) plays an important role in the pathogenesis of osteoarthritis. Schisandrin A, a bioactive compound found in fruits of the Schisandra genus, has been reported to possess multiple pharmacological and therapeutic properties. Although several studies have described the antioxidant effects of analogues of schisandrin A, the underlying molecular mechanisms of this bioactive compound remain largely unresolved. The present study investigated the cytoprotective effect of schisandrin A against oxidative stress (hydrogen peroxide [$H_2O_2$]) in SW1353 human chondrocyte cells. The results showed that schisandrin A preconditioning significantly inhibited $H_2O_2-induced$ growth inhibition and apoptotic cell death by blocking the degradation of poly (ADP-ribose) polymerase proteins and down-regulating pro-caspase-3. These antiapoptotic effects of schisandrin A were associated with attenuation of mitochondrial dysfunction and normalization of expression changes of proapoptotic Bax and antiapoptotic Bcl-2 in $H_2O_2-stimulated$ SW1353 chondrocytes. Furthermore, schisandrin A effectively abrogated $H_2O_2-induced$ intracellular ROS accumulation and phosphorylation of histone H2AX at serine 139, a widely used marker of DNA damage. Thus, the present study demonstrates that schisandrin A provides protection against $H_2O_2-induced$ apoptosis and DNA damage in SW1353 chondrocytes, possibly by prevention of ROS generation. Collectively, our data indicate that schisandrin A has therapeutic potential in the treatment of oxidative disorders caused by overproduction of ROS.

• Korean Journal of Food Science and Technology
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• v.46 no.3
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• pp.384-389
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• 2014

The objective of the present study was to investigate the chemical composition of anthocyanin-enriched extract of radiation-induced blackberry (Rubus fruticosus L.) mutant (${\gamma}$-B201) as well as the protective effect of ${\gamma}$-B201 against oxidative stress in vitro. The cytotoxicity, reactive oxygen species (ROS) scavenging capacity, and DNA damage were assessed by WST-1 assay, flow cytometry, and comet assay, respectively. Lactate dehydrogenase, superoxide dismutase, and catalase activities were determined by using a commercial kit. The in vitro results showed that ${\gamma}$-B201 increased the cell viability, reduction of lactate dehydrogenase release, and intracellular ROS scavenging capacity in hydrogen peroxide ($H_2O_2$)-treated HepG2 cells. Furthermore, treatment with ${\gamma}$-B201 attenuated DNA damage in $H_2O_2$-treated HepG2 cells and treatment with ${\gamma}$-B201 restored the activity of superoxide dismutase and catalase in $H_2O_2$-treated HepG2 cells. In conclusion, the present study suggests that ${\gamma}$-B201 blackberry extract can exert a significant cytoprotective effect against $H_2O_2$-induced cell damage.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.7
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• pp.953-959
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• 2010

We extracted bioactive materials from Codium fragile by enzymatic hydrolysis using four different proteases (Alcalase, Flavourzyme, Neutrase, and Protamex) and seven different carbohydrases (amyloglucosidase (AMG), Celluclast, Dextrozyme, Maltogenase, Promozyme, Termamyl, and Viscozyme), and evaluated their biological activities such as antioxidant, anti-acetylcholinesterase (AChE), and anti-inflammatory effects. All enzymatic hydrolysates showed good DPPH radical scavenging capacities, in particular, Flavourzyme and Promozyme hydrolysates possessed the highest activity. The two hydrolysates also exhibited strong hydrogen peroxide scavenging activity, $Fe^{2+}$ chelating activity, and reducing power in a dose-dependent manner. Furthermore, the two hydrolysates effectively protected DNA damage induced by hydroxyl radical by measuring the conversion of supercoiled DNA to the open circular DNA. All enzymatic hydrolysates also showed high anti-AChE inhibitory activities in a dose-dependent manner, and did not showed any significant cytotoxicity on RAW264.7 cells (p<0.05). In addition, the enzymatic hydrolysates significantly (p<0.05) inhibited lipopolysaccharide induced-nitric oxide production on RAW264.7 cells. These results suggest that the enzymatic extracts from Codium fragile would be good source as an ingredient of functional foods.

• Journal of Life Science
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• v.22 no.7
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• pp.936-942
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• 2012

Prunus mume has been traditionally used as a medicinal food in Korea, Japan, and China. In particular, this fruit has been reported to have beneficial biological effects on gastritis and gastric ulcers. However, its action in relation to skin whitening has remained unclear. Accordingly, the effects of fruit extract of P. mume related to antioxidation and skin whitening were examined in this study. First, using the MTT assay, it was observed that fruit extract of P. mume below 0.1% has no cytotoxicity in B16-F1 cells as a result of cell viability. Second, the direct scavenging effects and the reducing power of the fruit extract of P. mume were evaluated in vitro on DPPH radicals, hydrogen peroxide, and superoxide. It exhibited high reducing power and scavenging activity on the aforementioned reactive oxygen species. Furthermore, we found that its protective effect against genomic DNA damage related to oxidative stress was increased in a dose-dependent manner. In addition, the fruit extract of P. mume had an inhibitory effect on melanin production induced by L-dopa. In addition, it reduced the expression level of NRF-2, SOD-1, and SOD-2 related to antioxidation in western blot analysis. These results suggest that fruit extract of P. mume could exert a whitening effect through inhibition of melanin production by its antioxidant effect.