• Korean Journal of Environmental Biology
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• v.27 no.2
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• pp.230-236
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• 2009

An extract of Allomyrina dichotoma larva (ADL), one of the insects used most frequently in traditional Chinese medicine for the treatment of liver diseases such as hepatocirrhosis and hepatofibrosis, was assessed for antioxidant bioactivity in this study. In the current work, we have investigated the protective effects of ADL extracts on tert-butyl hydroperoxide (t-BHP)-induced hepatotoxicity in cultured hepa1c1c7 cells and in the mouse liver. The treatment of the hepa1c1c7 cells with ADL extracts induced a significant reduction of t-BHP-induced oxidative injuries, as determined by cell cytotoxicity, lipid peroxidation (LPO) and reactive oxygen species contents, in a dose-dependent manner. Moreover, ADL extracts evidenced a protective effect against t-BHPinduced oxidative DNA damage, as revealed by the results of the Comet assay in hepa1c1c7 cells. ADL extracts also protected against hydroxyl radical-induced 2-deoxy-d-ribose degradation by ferric ion-nitrilotriacetic acid and $H_2O_2$. In addition, ADL extracts were shown to be able to quench 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals. Our in vivo study revealed that ADL extracts pretreatment applied prior to t-BHP administration significantly prevented an increase in the serum levels of hepatic enzyme markers and reduced LPO in the mouse liver in a dose-dependent manner. Taken together, these results suggest that the protective effects of ADL extracts against t-BHP-induced hepatotoxicity may be attributable, at least in part, to its ability to scavenge free oxygen radicals, and to protect against DNA damage due to oxidative stress.

• The Journal of Korean Medicine
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• v.18 no.1
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• pp.375-384
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• 1997

This study was undertaken to determine Juglandis Semen extraction (JS) has a protective effect against the cell injury caused by oxidants, t-butylhydroperoxide (t-BHP) and $H_{2}O_2$ in rabbit lung slices. Cell injury was estimated by measuring tissue water content and peroxidation of membrane lipids was assessed by measuring malondialdehyde (MDA), an end-product of lipid peroxidation. t-BHP significantly increased water content in lung tissues over concentrations of 2-10 mM, and such effects were prevented by 5% JS. JS exerted the beneficial effect in a dose-dependent manner. $H_{2}O_2$ (100 mM) also increased water content in tissues, which was almost completely prevented by 5% JS. t-BHP induced lipid peroxidation in a dose-dependent fashion in lung tissues over concentrations of 0.5-10 mM. JS significantly reduced t-BHP induced lipid peroxidation and oxidant-independent endogenous lipid peroxidation, and such effects were dose-dependent at concentration of 0.5-10%. JS prevented $H_{2}O_2$ (100 mM)-dependent lipid peroxidation. These results suggest that JS prevents ceil injury induced by oxidants in the lung, and such effects may be attributed to inhibition of lipid peroxidation. The precise mechanisms remains to be explored.

• Biomedical Science Letters
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• v.10 no.4
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• pp.341-346
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• 2004

Tumor necrosis factor-α (TNF-α) or its mRNA expression are increased in acute nephrosis of various types including ischemia/reperfusion injury. This study was undertaken to determine whether pentoxifylline (PTX), an inhibitor of TNF-α production, provides a protective effect against hypoxia-induced cell injury in rabbit renal cortical slices. To induce hypoxia-induced cell injury, renal cortical slices were exposed to 100% N₂ atmosphere. Control slices were exposed to 100% O₂ atmosphere. The cell injury was estimated by measuring lactate dehydrogenase (LDH) release and p-aminohippurate (PAH) uptake. Exposure of slices to hypoxia increased the LDH release in a time-dependent manner. However, when slices were exposed to hypoxia in the presence of PTX, the LDH release was decreased. The protective effect of PTX was dose-dependent over the concentrations of 0.05∼1 mM. Hypoxia did not increase lipid peroxidation, whereas an organic hydroperoxide t-butylhydroperoxide (tBHP) resulted in a significant increase in lipid peroxidation. PTX did not affect tBHP-induced lipid peroxidation. Hypoxia decreased PAH uptake, which was significantly attenuated by PTX and glycine. tBHP-induced inhibition of PAH uptake was not altered by PTX, although it was prevented by antioxidant deferoxarnine. The PAH uptake by slices in rabbits with ischemic acute renal failure was prevented by PTX pretreatment. These results suggest that PTX may exert a protective effect against hypoxia-induced cell injury and its effect may due to inhibition of the TNF-α production, but not by its antioxidant action.

• The Journal of Internal Korean Medicine
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• v.21 no.2
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• pp.299-308
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• 2000

This study was performed to elucidate the effects of Gokajisilsosiho-Tang(GJST) on the lactic dehydrogenase(LDH) release, cell viability and activity, lipid peroxidation, DNA synthesis and the changes of total protein synthesis and GSH changes in vivo and in vitro in rat cultured hepatocytes from hydrogen peroxide$(H_2O_2)$-induced injury. The GJST extract had not an effect on cytotoxicity in all experimental results. The treatment of GJST extract of $160{\mu}g/ml$, $320{\mu}g/ml$ showed the significant effect to decrease LDH leakage induced by t-BHP in cultured rat hepatocytes. The higher concentration of GJST extract than $160{\mu}g/ml$, showed the inhibitory effect on decreasing cell viability induced by t-BHP. The treatment of t-BHP to rat cultured hepatocytes resulted in a concentration dependent increase in TBARS, in the presence of GJST extract the production of TBARS induced by hydrogen peroxide was inhibited concentration dependently, significantly inhibited at $80{\mu}g/ml$ of GJST extract and above. The GJST extract simutaneously present with t-BHP prevented the loss of total protein and GSH in a concentration dependent manner. These results suggested that GJST extract may play a hepatoprotective role in oxidative damage induced by hydrogen peroxide and a therapeutic potential of inhibiting liver injury.

• BMB Reports
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• v.40 no.6
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• pp.928-935
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• 2007

Three Angelica sinensis polysaccharide fractions (APFs), named APF1, APF2 and APF3, were isolated and purified from Radix A. sinensis and their antioxidant activities were evaluated in isolated mouse peritoneal macrophages by pretreatment with APFs before exposure to 0.2 mM tertbutylhydroperoxide (t-BHP). The results showed that pretreatment of the macrophages with APFs as low as $10{\mu}g$/ml could significantly enhance t-BHP-decreased cell survival, intracellular glutathione (GSH) content and superoxide dismutase (SOD) activity, and also inhibited t-BHP-increased lactate dehydrogenase (LDH) leakage and malondialdehyde (MDA) formation (p < 0.05), and APF3 was the most active fraction, followed by APF2 and APF1 in decreasing order. Furthermore, we found for the first time that the bound-protein in APF3 was associated closely with the protective effects and the polysaccharide inhibited the excess NO release from t-BHP-activated macrophages to protect host cells.

• BMB Reports
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• v.46 no.10
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• pp.513-518
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• 2013

We investigated the protective effects of Gymnaster koraiensis against oxidative stress-induced hepatic cell damage. We used two different cytotoxicity models, i.e., the administration of tert-butyl hydroperoxide (t-BHP) and acetaminophen, in HepG2 cells to evaluate the protective effects of G. koraiensis. The ethyl acetate (EA) fraction of G. koraiensis and its major compound, 3,5-di-O-caffeoylquinic acid (DCQA), exerted protective effects in the t-BHP-induced liver cytotoxicity model. The EA fraction and DCQA ameliorated t-BHP-induced reductions in GSH levels and exhibited free radical scavenging activity. The EA fraction and DCQA also significantly reduced t-BHP-induced DNA damage in HepG2 cells. Furthermore, the hexane fraction of G. koraiensis and its major compound, gymnasterkoreayne B (GKB), exerted strong hepatoprotection in the acetaminophen-induced cytotoxicity model. CYP 3A4 enzyme activity was strongly inhibited by the extract, hexane fraction, and GKB. The hexane fraction and GKB ameliorated acetaminophen-induced reductions in GSH levels and protected against cell death.

• Journal of Ginseng Research
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• v.36 no.3
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• pp.248-255
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• 2012

Potential antioxidant effect of processed ginseng (sun ginseng, SG) on oxidative stress generated by tert-butyl hydroperoxide (t-BHP) was investigated in HepG2 cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and lactate dehydrogenase (LDH) leakage test demonstrated that SG dose-dependently prevents a loss of cell viability against t-BHP-induced oxidative stress. Also, SG treatment dose-dependently relieved the increment of activities of hepatic enzymes, such as aspartate aminotrasferase and alanine aminotransferase, and lipid peroxidation mediated by t-BHP treatment in HepG2 cells. SG increased the gene expression of antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase. However, high dose of SG treatment caused decrease in mRNA level of glutathione peroxidase as compared to low dosage of SG-treated cells. The gene expression of glutathione reductase was found to be slightly increased by SG treatment. In addition, SG extract attributed its hepaprotective effect by inducing the mRNA level of bcl-2 and bcl-xL but reducing that of bax. But, the gene expression of bad showed no significant change in SG-treated HepG2 cells. These findings suggest that SG has hepatoprotective effect by showing reduction of LDH release, activities of hepatic enzymes and lipid peroxidation and regulating the gene expression of antioxidant enzymes and apoptosis-related molecules against oxdative stress caused by t-BHP in HepG2 cells.

• Preventive Nutrition and Food Science
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• v.21 no.3
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• pp.208-220
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• 2016

The aim of this study was to investigate the protective effects of juice powders from sweet orange [Citrus sinensis (L.) Osbeck], unshiu mikan (Citrus unshiu Marcow), and mini tomato (Solanum lycopersicum L.), and their major flavonoids, hesperidin, narirutin, and rutin in tert-butyl hydroperoxide (t-BHP)-induced oxidative stress in HepG2 cells. The increased reactive oxygen species and decreased glutathione levels observed in t-BHP-treated HepG2 cells were ameliorated by pretreatment with juice powders, indicating that the hepatoprotective effects of juice powders and their major flavonoids are mediated by induction of cellular defense against oxidative stress. Moreover, pretreatment with juice powders up-regulated phase-II genes such as heme oxygenase-1 (HO-1), thereby preventing cellular damage and the resultant increase in HO-1 expression. The high-performance liquid chromatography profiles of the juice powders confirmed that hesperidin, narirutin, and rutin were the key flavonoids present. Our results suggest that these fruit juice powders and their major flavonoids provide a significant cytoprotective effect against oxidative stress, which is most likely due to the flavonoid-related bioactive compounds present, leading to the normal redox status of cells. Therefore, these fruit juice powders could be advantageous as bioactive sources for the prevention of oxidative injury in hepatoma cells.

• Journal of Sasang Constitutional Medicine
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• v.13 no.1
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• pp.51-60
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• 2001

Effects of Taeumjowetang on Lipid Peroxidation by Free Radicals and Oxidative Damage of Hepatocytes by tert-Butyl Hydroperoxide. 1. Purpose The present study was carried out to evaluate the antioxidant effects of Taeumjowetang in vitro. 2. Methods In this study, antioxidant effects of TJT on lipid peroxidation were determined according to the method of TBA. (Abbreviation) TJT : Taeumjowetang, TBA : 2-thiobarbituric acid. 3. Results : 1) TJT inhibited markedly peroxidation of linoleic acid during the autoxidation. 2) TJT inhibited lipid peroxidation induced by hydroxyl radical derived from H2O2-Fe2+ in rat liver homogenate. 3) TJT showed 66% scavenging effect on DPPH radical. 4) TJT exhibited a 25% inhibitory effect on superoxide generation from xanthine-xan thine oxidase system. 5) To investigate the antioxidative effects of TJT on the hepatocytes, cultured normal rat liver cells(Ac2F) were prepared and incubated with or without TJT. After 16~18hr, cells placed in DMEM medium without serum, and then incubated with 1mM t-BHP for 2hr. Viable cells were detected by MTT assay. In this test, TJT protected the cell death induced by t-BHP and significantly increased cell viability in the normal rat liver cell. (Abbreviation) DPPH : ${\alpha},{\alpha}$-diphenyl-${\beta}$-picryl hydrazyl, DMEM : Dulbecco's Modified Eagle Medium, t-BHP : terr-butyl hydroperoxide, 4. Conclusion These results suggested that TJT might play a protective role in lipid peroxidation by free radicals.

• Animal Bioscience
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• v.36 no.7
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• pp.1120-1129
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• 2023

Objective: This study aimed to determine the protective efficacy of Buddha's Temple (BT) extract against tert-butyl hydroperoxide (t-BHP)-induced oxidative stress in Gallus gallus chicken embryo fibroblast cell line (DF-1) and its effects on the cell lipid metabolism. Methods: In this experimental study, Gallus gallus DF-1 fibroblast cells were pretreated with BT 10^-7 for 24 hours, followed by their six-hour exposure to t-BHP (100 μM). Water-soluble tetrazolium salt-8 (WST-8) assays were performed, and the growth curve was computed. The intracellular gene expression changes caused by BT extract were confirmed through quantitative polymerase chain reaction (qPCR). Flow cytometry, oil red O staining experiment, and thin-layer chromatography were performed for the detection of intracellular metabolic mechanism changes. Results: The WST-8 assay results showed that the BT pretreatment of Gallus gallus DF-1 fibroblast cell increased their cell survival rate by 1.08%±0.04%, decreased the reactive oxygen species (ROS) level by 0.93%±0.12% even after exposure to oxidants, and stabilized mitochondrial activity by 1.37%±0.36%. In addition, qPCR results confirmed that the gene expression levels of tumor necrosis factor α (TNFα), TIR domain-containing adapter inducing IFN-beta (TICAM1), and glucose-regulated protein 78 (GRP78) were regulated, which contributed to cell stabilization. Thin-layer chromatography and oil red O analyses showed a clear decrease in the contents of lipid metabolites such as triacylglycerol and free fatty acids. Conclusion: In this study, we confirmed that the examined BT extract exerted selective protective effects on Gallus gallus DF-1 fibroblast cells against cell damage caused by t-BHP, which is a strong oxidative inducer. Furthermore, we established that this extract significantly reduced the intracellular ROS accumulation due to oxidative stress, which contributes to an increase in poultry production and higher incomes.