• Korean Journal of Pharmacognosy
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• v.35 no.1 s.136
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• pp.28-34
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• 2004

DNA damage induced by reactive oxygen species (ROS) seems to play an important role in the induction of mutation and cancer. Hydrogen peroxide $(H_2O_2)$ has been shown to induce a variety of genetic alterations, probably by the generation of hydroxyl radicals via Fenton reaction. In this study, we examined the ability of medicinal herbs in the suppression of $H_2O_2$-induced mutagenesis. Human fibroblast GM00637 cells were treated with $H_2O_2$ in the presence or absence of medicinal herbs, and $H_2O_2$-induced mutant frequency was measured at the hypoxanthine guanine phosphoribosyl transferase (HPRT) locus. Treatment of cells with various doses of $H_2O_2$ caused a significant increase of the HPRT mutant frequency. However, pretreatment of cells with several medicinal herbs reduced $H_2O_2$-induced mutant frequency. The strong antimutagenic effects were observed from the methylene chloride and ethyl acetate fractions of Selaginella tamariscina, Panax ginseng, and Angelica acutiloba; ethyl acetate fractions of Rehmania glutinosa, Leonurus sibiricus, Curcuma zedoaria and Commiphora molmol; butanol fractions of Scutellaria barbata, Tribulus terrestris, Curcuma zedoaria, Cyperus rotundus and Carthamus tinctorius, which were more than 60% inhibition of $H_2O_2$-induced mutant frequency at the HPRT locus.

• The Journal of Korean Medicine
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• v.21 no.4
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• pp.193-203
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• 2000

Objectives : Hindered barrier function of vascular endothelium has been implicated in the initiation and progression of degenerative vascular diseases such as atherosclerosis. In this study, the effect of Sunghyangchungisan(SHCS) as a protectant against oxidant-induced destruction of endothelial barrier function was assessed. Methods : Toward this end, endothelial cells derived from the human umbilical vein were cultured as monolayers on permeable membrane filters. Endothelial permeability was monitored by measuring transendothelial electrical resistance and movement of low density lipoprotein (LDL) across the endothelial monolayer. Results : Along with increased movement of LDL, $H_2O_2$-induced increase in endothelial permeability was paralleled by a decrease in transendotheliaI electrical resistance. The effect of $H_2O_2$ was mimicked by phorbol 12-myristate 13-acetate (PMA), a potent activator of proteinkinase C. Calphostin-C, a protein kinase C inhibitor, effectively blocked the increase in endothelial permeability induced by $H_2O_2$ or PMA, indicating that activation of protein kinase C is associated with the $H_2O_2-induced$ permeability change. SHCS effectively protected the endothelial monolayer against $H_2O_2-induced$ increase in permeability, whereas, it did not affect PMA-induced change. Forskolin, a potent activator of adenylyl cyclase, antagonized $H_2O_2$ to increase endothelial permeability. In addition, in ${H_2O_2}-treated$ cens, intracenular cAMP concentration was significantly decreased, indicating that impaired cAMP production as well as activation of proteinkinase C is a mechanism underlying ${H_2O_2}>-induced$$H_2O_2$ with regard to its effect on intracellular cAMP content. However, SHCS itself did not affect resting cAMP concentration in endothelial cells. Conclusions : These results suggest that SHCS might operate as an effective protectant against oxidant-induced destruction of endothelial barrier function. The mechanism does not appear to involve direct interaction with protein kinase C- or cAMP-associated signaling mechanism.

• Korean Journal of Clinical Laboratory Science
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• v.43 no.4
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• pp.161-170
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• 2011

Quercetin is one of the most distributed flavonoids in the plant kingdom and occurs naturally in a wide range of fruits and vegetables. This study was undertaken to determine whether quercetin exerts beneficial effect against necrotic and apoptotic cell death induced by hydrogen peroxide ($H_2O2$) in intestinal cells using the human-derived cultured T84 colonic epithelial cell line. Necrotic cell death was induced by exposing cells to 0.5 mM $H_2O_2$ for 2 h and apoptosis was induced by incubating cells in normal culture medium for 18 h following exposure of cells to 0.5 mM $H_2O2$ for 2 h. Cell viability was evaluated by the trypan blue exclusion assay and apoptosis was assessed by Hoechst 33258 staining and flow cytometry. $H_2O_2$ induced necrotic cell death in a time and dose-dependent fashion. Both necrotic and apoptotic cell deaths were not prevented by the antioxidants N,N'-diphenyl-p-phenylenediamine(DPPD) and Trolox, whereas both cell deaths induced by the organic hydroperoxide t-butylhydroperoxide (tBHP) were prevented by DPPD, suggesting that $H_2O_2$ induces cell death through a lipid peroxidation-independent mechanism. $H_2O2$-induced necrotic death was prevented by deferoxamine and 3-aminobenzamide, while the apoptotic cell death was not affected by these agents. Quercetin prevented both necrotic and apoptotic cell deaths induced by $H_2O_2$ in a dose-dependent manner. $H_2O_2$ caused activation of poly (ADP-ribose) polmerase (PARP), which was inhibited by deferoxamine, 3-aminobenzamide, and quercetin, but not DPPD. These results indicate that quercetin inhibits both necroticand apoptotic deaths of T84 cells. The anti-necrotic effect of quercetin may be attributed to its iron chelator activity rather than a direct $H_2O_2$ scavenging capacity and antioxidant. The present study suggests that quercetin may play a therapeutic role in the treatment of human gastrointestinal diseases mediated by oxidants.

• The Journal of Internal Korean Medicine
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• v.20 no.1
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• pp.83-98
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• 1999

This study is designed to investigate the effects of Samul-Tang extract on the response of lactic dehydrogenase(LDH) release, cellular activity, lipid peroxidation, DNA synthesis and the changes of total protein of bovine pulmonary artery endothelial cells(PAEC) from hydrogen peroxide$(H_2O_2)$-induced injury. The results are as follows : 1. Samul-Tang significantly decreased $H_2O_2$-induced release of LDH from injured bovine PAEC. 2. Samul-Tang significantly repressed $H_2O_2$-induced cellular activity from injured bovine PAEC. 3. Samul-Tang significantly repressed $H_2O_2$-induced lipid peroxidation from injured bovine PAEC. 4. Samul-Tang significantly stimulated DNA synthesis in bovine PAEC. 5. Samul-Tang significantly repressed $H_2O_2$-induced changes of total protein volume from injured bovine PAEC. Above results suggest that Samul-Tang can protect bovine PAEC from $H_2O_2$-induced injury. These results can be effectively applied to the prevention and cure of cardiovascular and cerebrovascular diseases.

• The Journal of Korean Medicine
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• v.22 no.4
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• pp.58-68
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• 2001

Objectives : This study was designed to investigate the protective mechanisms of Samul-tang (SMT) on $H_2O_2$-induced toxicity in H9c2 cardiomyoblast cells. Methods : The cultured cells were pretreated with SMT and exposed to $H_2O_2$. The cell damage was assessed by using MTT assay. Also, we used Hoechst staining, Western blotting analysis. Results : SMT significantly reduced both $H_2O_2$-induced cell death and chromatin fragmentation. The decrease of Bcl2 expression by $H_2O_2$ was inhibited by SMT. In addition, the increase of Bax expression was also inhibited by SMT. In particular, Fas expression, which is generally recognized as cell death inducing signal by Fas/FasL interaction, was markedly decreased by $H_2O_2$ in a time-dependent manner, whereas this decrease was completely prevented by SMT. The cotreatment of SMT and $H_2O_2$ in H9c2 cells also induced the phosphorylation of ERK in a time-dependent manner. Moreover, PD098059, a specific inhibitor of ERKl/2, attenuated the protective effect of SMT on $H_2O_2$-induced toxicity in H9c2 cardiomyoblast cells. Furthermore, the protective effect of SMT was significantly blocked by treatment of SB203580, a specific inhibitor of p38. Conclusions : Taken together, this study suggests that the protective effects of the water extract of SMT against oxidative damages may be mediated by the modulation of Bel2 and Bax expression via the regulation of ERK and p38 signaling pathway.

• The Korean Journal of Physiology and Pharmacology
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• v.13 no.3
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• pp.139-145
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• 2009

The purpose of this study was to evaluate the effects of the water extract of Samultang (SMT), a Chinese herb, on apoptotic cell death by $H_2O_2$-induced oxidative stress in SK-N-M C cells. A nuclear fragmentation was observed via fluorescence imaging 12 h after exposure to 30 ${\mu}M$ $H_2O_2$ and DNA laddering was detected via agarose electrophoresis gel. In addition, increases in sub-G1 phase and cleavage of the PARP protein were observed. However, treatment with SMT for 2 h prior to $H_2O_2$ exposure significantly reduced apoptotic cell death induced by incubation with 30 ${\mu}M$ $H_2O_2$ in SK-N-MC cells. Pre-incubation with water extract of SMT for 2 h prevented the $H_2O_2$-induced decrease in mitochondrial transmembrane potential. SMT also attenuated the increase in caspase-3 activity and the breakdown of PARP protein caused by $H_2O_2$-induced oxidative stress. These results suggest that the water extract of SMT provides inhibition of apoptotic cell death against oxidative injury in SK-N-MC cells.

• Bulletin of the Korean Chemical Society
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• v.27 no.6
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• pp.830-834
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• 2006

Lipid peroxidation induced by the reaction of cytochrome c with $H_2O_2$ was investigated. When linoleic acid micelles or phosphatidyl choline liposomes were incubated with cytochrome c and $H_2O_2$, lipid peroxidation was increased in cytochrome c and $H_2O_2$ concentrations-dependent manner. Radical scavengers, azide, formate and ethanol prevented lipid peroxidation induced by the cytochrome c/$H_2O_2$ system. Iron specific chelator, desferoxamine also prevented the cytochrome c/$H_2O_2$ system-mediated lipid peroxidation. These results suggest that lipid peroxidation may be induced by the cytochrome c/$H_2O_2$ system via the generation of free radicals. Carnosine, homocarnosine and anserine are present in the muscle and brain of many animals and human. Previous studies show that these compounds have an antioxidant function. In the present study, carnosine, homocarnosine and anserine significantly prevented the cytochrome c/$H_2O_2$ system-mediated lipid peroxidation. Carnosine and related compounds also inhibited the free radical-generating activity of cytochrome c. The results suggest that carnosine, homocarnosine and anserine may prevent lipid peroxidation induced by the cytochrome c/$H_2O_2$ system through a free radical scavenging.

• 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.

• Biomedical Science Letters
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• v.14 no.3
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• pp.193-198
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• 2008

In order to evaluate on the effect of kaempferol on the cytotoxicity of oxygen tree radicals, XTT assay was performed to determine the cell viability after skin fibroblasts derived from human (Detroit 51) that were treated with various concentrations of hydrogen peroxide $(H_2O_2)$. And also, the effect of kaempferol on the cytotoxicity induced by H202 that was examined by cell viability, lactate dehydrogenase (LDH) activity and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity in these cultures. $H_2O_2$ decreased cell viability in dose-dependent manner in these cultures and the $XTT_{90}\;and\;XTT_{50}$ values were determined at concentration of $35{\mu}M\;and\;90{\mu}M$ of $H_2O_2$ after skin fibroblasts derived from human were treated with $15{\sim}90{\mu}M$ of $H_2O_2$ for 6 hours, respectively. $H_2O_2$ was highly toxic on cultured skin fibroblasts derived from human by toxic criteria of Brenfreund and Puerner (1984). In the protective effect of kaempferol on $H_2O_2$-induced cytotoxicity, kaempferol increased DPPH radical scavenging activity and significantly decreased LDH activity. From these results, it is suggested that oxygen tree radical, $H_2O_2$, was highly toxic on cultured skin fibroblasts derived from human, and also kaempferol of flavonoid showed the protection on $H_2O_2$-induced cytotoxicity.

• Biomedical Science Letters
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• v.14 no.1
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• pp.27-32
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• 2008

To clerify the antioxidant effect of Crataegi Fructus (CF) extract on reactive oxygen species (ROS), The C6 glioma cells were treated with various concentrations of hydrogen peroxide ($H_2O_2$). The $H_2O_2$-induced neurotoxicity was measured by XTT assay for the cell viability. For the protective effect of CF extract on the cytotoxicity induced by $H_2O_2$, cell viability, lactate dehydroganase (LDH) activity, and the inhibitive activity of lipid peroxidation of CF extract were performed. In this study, $H_2O_2$ decreased cell viability dose- and time-dependent manners and increased LDH activity compared with the control. In the protective effect on $H_2O_2$, CF extract increased cell viability and decreased LDH activity on $H_2O_2$-induced cytotoxicity, lipid peroxidation by FTC assay. From these results, It is suggested that $H_2O_2$ was highly toxic on cultured C6 glioma cells, and also, CF extract showed the protective effect on $H_2O_2$-mediated cytotoxicity.