• Journal of Acupuncture Research
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• v.17 no.3
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• pp.176-187
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• 2000

Objectives : This study was purposed to investigate the antioxidative effects of Paeoniae radix aqua-acupuncture solution(PR) on culture liver cell system, lipid peroxidation and antioxidative enzyme activities in tert-butyl hydroperoxide(t-BHP) treatmented conditions. Methods : Cultured normal rat liver cell(Ac2F) were prepared and incubated with or without PR(at 2% volume in culture medium). 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, and the levels of lipid peroxide(LPO) were measured by TBA method. And catalase activity was measured as the decrease in hydrogen peroxide absorbance at 240nm on spectrophotometer using 30mM hydrogen peroxide. Superoxide dismutase(SOD) were assayed by recording the inhibition of nitro blue tetrazolium reduction with xanthine and xanthine oxidase. Glutathione peroxidase(GPX) activity was determined by the modified coupled assay developed by Paglia and Lawrence. The reaction was started by addition of 2.2mM hydrogen peroxide as substrate. The change in absorbance at 340nm was measured for 1min on spectrophotometer. Glutathione-S-transferase(GST) activity was assayed with CDNB as substrate and enzyme activity of GST towards the glutathione conjugation of CDNB. Results : Cell killing was significantly enhanced by addition of t-BHP compared to those of untreated group. PR pretreated cell resisted the toxic effects of t-BHP. LPO levels of t-BHP treatment group were significantly higher than other groups. This increased level was significandy reduced by PR pretreatment. The t-BHP treatment resulted in a decrease of catalase, GPX and GST activities. By contrast, PR pretreatment markedly increased compare to those of untreated groups. Conclusions : T-BHP which can produce intracellular free radical was used for inducer of the peroxidation of cellular lipids. PR protected the cell death induced by t-BHP and significantly increased cell viabiliry in the normal rat liver cell, and showed effective inhibition of lipid peroxidation, and elevations of catalase, GPX and GST activities. These results suggested that PR might play a protective role in lipid peroxidation by free radicals.

• Biomolecules & Therapeutics
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• v.19 no.2
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• pp.195-200
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• 2011

Neuronal cell death is a common characteristic feature of a variety of neurodegenerative disorders including Alzheimer's disease and Parkinson's disease. However, there have been no effective drugs to successfully prevent neuronal death in those diseases. In the present study, docosyl cafferate (DC), a derivative of caffeic acid, was isolated from Rhus verniciflua and its protective effects on tBHP-induced neuronal cell death were examined in SH-SY5Y human neuroblastoma cells. Pretreatment of DC significantly attenuated tBHP-induced neuronal cell death in a concentration-dependent manner. DC also significantly suppressed tBHP-induced caspase-3 activation. In addition, DC restored tBHP-induced depletion of intracellular Bcl-2, an anti-apoptotic member of the Bcl-2 family. Furthermore, DC significantly suppressed tBHP-induced degradation of IKB, which retains $NF-{\kappa}B$ in the cytoplasm, resulting in the suppression of nuclear translocation of $NF-{\kappa}B$ and its subsequent activation. Taken together, the results clearly demonstrate that DC exerts its neuroprotective activity against tBHP-induced oxidative stress through the suppression of nuclear translocation of $NF-{\kappa}B$.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.5
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• pp.1304-1308
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• 2008

This study was performed to evaluate the effect of persimmon leaves extract on the reactive oxygen species (ROS) in cultured melanoma cells. The B16/F10 melanoma cells were treated with various concentrations of t-butyl hydroperoxide (t-BHP). And also, the effect of persimmon leaves (PL) extract on the cytotoxicity mediated by t-BHP was done on the cell viability, tyrosinase activity and melanogenesis by colorimetric assays. In this study, t-BHP decreased cell viability in dose-dependent manner and XTT90 and XTT50 values were measured at 10 and 35 uM of PL, respectively in these culture. And also, XTT50 value was assessed as a highly toxic effect on cultured melanoma cells by the toxic criteria. In the effect of PL extract on the t-BHP-mediated cytotoxicity, PL extract significantly increased the cell viability injured by t-BHP in cultured B16/F10 melanoma cells. PL also showed the decreased tyrosinase activity and melanogenesis. From these results, it is suggested that ROS such as t-BHP showed highly toxic effect on cultured melanoma cells, and also, PL extract inhibited the tyrosinase activity and melanogenesis in cultured melanoma cells injured by ROS.

• Journal of Pharmacopuncture
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• v.10 no.1 s.22
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• pp.121-135
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• 2007

Objectives : This study was carried out to examine protective effect of wild ginseng extract on HepG2 human hepatoma cell line against tert-Butyl hydroperoxide (t-BHP)-induced oxidative damage. Methods : To evaluate protective effect of wild ginseng extract against t-BHP induced cytotoxicity, LDH level and activity of glutathione peroxidase and reductase were measured. Gene expression was also measured using DNA microarray. Results : Wild ginseng extract showed a significant protective effect against t-BHP-induced cytotoxicity in HepG2 cell line. It is not, however, related with the activities of glutathione peroxidase and glutathione reductase. Analysis of gene expression using DNA chip, demonstrated that 28 genes were up-regulated in t-BHP only group. Five genes - selenoprotein P, glutathione peroxidase 3, sirtuin 2, peroxiredoxin 2, serfiredoxin 1 homolog - may be related with the protective effect of wild ginseng extract. Conclusions : Based on the results, a protective effect of wild ginseng extract against t-BHP-induced oxidative damage in HepG2 cell line is not associated with the activities of glutathione peroxidase and glutathione reductase, but with the expression of selenoprotein P, glutathione peroxidase 3, sirtuin 2, peroxiredoxin 2, and serfiredoxin 1 homolog.

• Food Science and Preservation
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• v.24 no.1
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• pp.107-115
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• 2017

The aim of this study was to investigate the hepatoprotecive effect of silk protein hydrolysates (SDH), which was prepared by acid hydrolysis, in rats. SDH itself did not exhibit any cytotoxic effect on hepatic tissues. SDH showed a protective effect on tert-butyl hydroperoxide (t-BHP)-induced hepatotoxicity and liver damage. SDH effectively reduced AST (aspartate aminotransferase) and ALT (alanine aminotransferase), which are biomarkers for liver damage, in a dose-dependent manner. Malondialdehyde (MDA), a lipid peroxidation product, was significantly reduced by SDH. A high dose of SDH (2 g/kg) reduced t-BHP-induced MDA production by 40%. Glutathione (GSH), which is an endogenous antioxidant molecule, was effectively increased by SDH treatment. GSH content was enhanced by around 2.5-fold, compared with t-BHP control, upon SDH (2 g/kg) treatment. Lactate dehydrogenase (LDH), which is an enzyme released by cell cytotoxicity, was greatly increased by t-BHP, but significantly decreased by SDH treatment. Furthermore, hematoxylin and eosin (H&E) staining showed that SDH suppressed t-BHP-induced lesions in liver tissue. Taken together, SDH might be used as a protective agent against liver damage.

• Journal of Pharmacopuncture
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• v.2 no.1 s.2
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• pp.13-25
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• 1999

This study was undertaken to determine whether Juglandis Semen (JAS) extraction exerts protective effect against oxidant-induced inhibition of $Na^+$-pump activity in cerebral cortex. $Na^+$-pump activity was estimated by measuring ouabain-sensitive oxygen consumption. The oxygen consumption significantly inhibited by 1 mM t-butylhydroperoxide (tBHP), which was prevented by addition of 2% JAS extraction. The oxygen consumption was increased by an increase in $Na^+$ concentration from 5 to 100mM, $K^+$ concentration from 0.5 to 10 mM, and $Mg^{++}$ concentration from 0.2 to 5 mM. These changes in ion concentrations did not affect the inhibitory effct of tBHP and protective action of JAS on oxygen consumption. tBHP(1mM) produced a significant increase in lipid peroxidation in cerebral cortex, which was prevented by 2% JAS extraction. These result suggest that JAS exerts protective effect against tBHP-induced inhibition of $Na^+$-pump activity in the cerebral cortex, this effect may be due to by an antioxidant action.

• The Journal of Dong Guk Oriental Medicine
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• v.7 no.1
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• pp.87-98
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• 1998

This study was undertaken to determine whether Bombycis Corpus extract (Bom) has antioxidant action. Kidney tissues were exposed to t-butylhydroperoxide (t-BHP) to induce oxidative stress. Lipid peroxidation was estimated by measuring malondialdehyde, a product of lipid peroxidation, and cell injury was estimated by measuring lactate dehydrogenase (LDH) release in rabbit renal cortical slices. t-BHP increased lipid peroxidation and LDH release in a dose-dependent manner over the concentration range of 0.1-1 mM. Such effects of t-BHP on lipid peroxidase and LDH release were prevented by 0.5% Bom. When tissues were treated with t-BHP in the presence of various concentrations of Bom, lipid peroxidation and LDH release were dose-dependently inhibited by Bom. Bom at 1 and 2% concentrations inhibited lipid peroxidation and LDH release in normal tissues. Bom at 2% concentration increased glutathione peroxidase activity in tissues treated or untreated with 1.0 mM t-BHP. However, catalase activity was not altered by addition of Bom. Bom inhibited generation of reactive oxygen species. These results indicate that Bom inhibits lipid peroxidation and cell injury in tissues treated with or without oxidant and this effect is, at least in part, attributed to increased activity of glutathione peroxidase and a direct sacvenging action.

• Journal of Microbiology and Biotechnology
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• v.15 no.4
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• pp.887-890
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• 2005

To evaluate the hepatoprotective activity of lactic acid bacteria, their effects on tert-butylperoxide (t-BHP)-induced hepatotoxicity in mice were measured. When lactic acid bacteria at doses of 0.5 and 2 g (wet weight)/kg were orally administered to mice with t-BHP-induced liver injury, these bacteria significantly inhibited the increase of plasma alanine aminotransferase and aspartate aminotransferase activities by $17-57\%$ and $57-66\%$ of the t-BHP control group, respectively. However, these lactic acid bacteria did not protect cytotoxicity induced by t-BHP against HepG2 cells. The inhibitory effects of these lactic acid bacteria at a dose of 15 g/kg were comparable with that of diphenyl dimethyl bicarboxylate at a dose of 0.2 g/kg, which has been used as a commercial hepatoprotective agent. Among these lactic acid Jacteria, Bifidobacterium longum HY8001 exhibited the most potent hepatoprotective effect. These orally administered lactic acid bacteria inhibited liver lipid peroxidation on t-BHP-induced hepatotoxicity of mice. We suggest that lactic acid bacteria may be an effective agent against liver injury.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.288.1-288.1
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• 2002

Herbal medicines are increasingly being utilized to treat a wide variety of disease processes. The aim of this study was to evaluate the ability of aqueous extract from the roots of Platycodon grandiflorum A. DC (Campanulaceae). Changkil (CK). to affect cellular response in primary cultures of rat hepatocytes to t-butyl hydroperoxide (t-BHP) induced oxidative stress and hepatotoxicity. CK-treated cells showed an increased resistance to oxidative challenge. as revealed by a higher percent of survival capacity in respect to control cells. CK added prior or simultaneously with I-BHP reduced enganced lipid peroxidation measured as production of malondialdehyde and enhnaced intracellular reduced glutathinoe depletion by t-BHP. Furhtermore. CK protected from the t-BHP-induced intracellular generation of reactive oxygen species assessde by montioting dichlorodihydrofluorescein fluorescence. it can be concluded that CK exerts an antioxidant action insice the cell. responsible for the abserved modulation of the cellular response to oxidative challenge. and CK have a marked anitioxdative and hepatoprotective potency.

• Korean Journal of Acupuncture
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• v.17 no.1
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• pp.179-190
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• 2000

This study was undertaken to determine whether Juglandis semen extract solution (JLS solution) exerts protective effect against oxidant-induced inhibition of glutamate uptake by synaptosomes. Synaptosome was prepared from rabbit brain cortex. Glutamate uptake increased by incubation time during 10 minutes, which was significantly inhibited by 1mM t-buthylhydroperoxide(t-BHP). JLS solution prevented t-BHP-induced inhibition of glutamate uptake in a dose-dependent manner. t-BHP reduced glutamate uptake in dose-dependent fashion, which was significantly prevented by 2% JLS solution. t-BHP(1mM) and $ascorbate/Fe^{2+}(50/1{\mu}M)$ increased lipid peroxidation in synaptosomes by 5-fold, and it was significantly prevented by 2% JLS solution. $HgCl_2(0.1mM)$ inhibited glutamate uptake and increased lipid peroxidation. These changes were prevented by 2% JLS solution. Synaptosomal Na-K-ATPase activity was inhibited by t-BHP(1mM) and $H_2O_2(50mM)$, which was prevented by 2% JLS solution. The results indicate that JLS solution prevents oxidant-induced inhibition of glutamate by synaptosomes, and this may result from inhibition of lipid peroxidation induced by oxidants.