• The Journal of Korean Medicine
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• v.17 no.1 s.31
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• pp.9-20
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• 1996

Oxygen free radicals can generated during metabolic processes in normal cells and by exposure of cells to toxic substances. These radicals have been recogenized to playa critical role in several pathological conditions including carcinogenesis and aging, and they have been implicated in pathogenesis of various diseases such as seizure, Alzheimer's disease, Parkinson's disease, myocardial infarction, respiratory distress syndrome, and rheumatoid arthritis. This study was undertaken to determine if Juglandis semen herb-acupuncture solution (JSHAS) has a protective effect against cell injury caused by oxidants, t-butylhydroperoxide (t-BHP) and $H_{2}O_2$. Cell injury was estimated by measuring lactate dehydrogenase (LDH) release and lipid perexidation was estimated by measurimg malondialdehyde, a product of lipid peroxidation. JSHAS significantly prevented LDH release induced by t-BHP or $H_{2}O_2$ in a dose-dependent manner at concentrations of 0.5-10%. Such protective effect was observed in control tissues untreated with oxidants. JSHAS, at 5% concentration, significantly reduced LDH release even when the concentrations of t-BHP and $H_{2}O_2$ increased to 5 and 200 mM, respectively. JSHAS, at 5% concentration, significantly reduced the lipid peroxidation by t-BHP and $H_{2}O_2$. These results indicate that JSHAS prevents cell injury and lipid peroxidation induced by oxidants in rabbit kidney cells. However, the underlying mechanisms remain to determined.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.5
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• pp.601-609
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• 1998

The present study was undertaken to examine the role of phospholipase $A_2\;(PLA_2)$ in oxidant-induced inhibition of phosphate transport in primary cultured rabbit renal proximal tubule cells. Uptakes of phosphate and glucose were dose-dependently inhibited by an oxidant t-butylhydroperoxide (tBHP), and the significant inhibition appeared at 0.025 mM of tBHP, whereas tBHP-induced alterations in lipid peroxidation and cell viability were seen at 0.5 mM. tBHP stimulated arachidonic acid (AA) release in a dose-dependent fashion. A $PLA_2$ inhibitor mepacrine prevented tBHP-induced AA release, but it did not alter the inhibition of phosphate uptake and the decrease in cell viability induced by tBHP. tBHP-induced inhibition of phosphate transport was not affected by a PKC inhibitor, staurosporine. tBHP at 0.1 mM did not produce the inhibition of $Na^+-K^+-ATPase$ activity in microsomal fraction, although it significantly inhibited at 1.0 mM. These results suggest that tBHP can inhibit phosphate uptake through a mechanism independent of $PLA_2$ activation, irreversible cell injury, and lipid peroxidation in primary cultured rabbit renal proximal tubular cells.

• 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.

• 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 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.

• The Korea Journal of Herbology
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• v.26 no.3
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• pp.47-56
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• 2011

Objectives : This study investigated whether the water extract of Spatholobi Caulis (SCE) has the ability to protect hepatocyte against oxidative stress induced by tert-butylhydroperoxide (tBHP) in vitro and $CCl_4$ in vivo. Methods : In vitro, HepG2 cells pre-treated with Spatholobi Caulis water extract (1, 3, 10, $30{\mu}g$/ml) for 12h and further incubated with tBHP ($100{\mu}M$) for the next 12h. Cell viability was assessed by MTT assay. In vivo, rats were orally administrated with the aqueous extract of Spatholobi Caulis (SCE; 50, 100 mg/kg) for 4 days and then, injected with $CCl_4$ 1 mg/kg body weight to induce acute liver damage. Results : Treatment with SCE inhibited cell death induced by tBHP, as evidenced by alterations in the levels of the proteins associated with apoptosis:SCE prevented a decrease in $Bcl_2$, and cleavage of poly(ADP-ribose)polymerase and pro-caspase-3. Moreover, SCE inhibited the ability of tBHP to generate $H_2O_2$ production, thereby restoring GSH content. Moreover, SCE treatments in rats effectively decreased liver injuries induced by a single dose of $CCl_4$, as evidenced by decreases in hepatic degeneration and inflammation as well as plasma alanine aminotransferase and lactate dehydrogenase activities. Consistently, treatments of SCE also protected liver in rats stimulated by $CCl_4$, as indicated by restoration GSH and prevention of MDA in the liver. Conclusions : SCE has the ability 1) to protect hepatocyte against oxidative stress induced by tBHP and 2) to prevent $CCl_4$-inducible acute liver toxicity. Present findings may be informative not only in elucidating the pharmacological mechanism of Spatholobi Caulis, but in determining its potential application for oxidative cellular damage in the liver.

• Korean Journal of Plant Resources
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• v.23 no.2
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• pp.165-171
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• 2010

Oxidative stress induced by reactive oxygen intermediates has been implicated in a variety of human diseases including neurodegenerative disorders, cancer, cardiovascular and respiratory diseases, and mode of action of environmental toxicants. Tert-butylhydroperoxide (t-BHP) is an organic lipid hydroperoxide analogue, which is commonly used as a pro-oxidant for evaluating mechanisms involving oxidative stress in cells and tissues. In this study, the underlying mechanisms involved in the protective effects of Hwabuk 94 kiwifruit (Actinidia deliciosa cv. 'Hwabuk 94'), which is cultivated in Jeju, on the t-BHP-induced cytotoxicity in PC12 cell. The pretreatment of rat pheochromocytoma cell line PC12 with Hwabuk 94 extract ($1-100\;{\mu}g/ml$) resulted in a significant recovery from t-BHP-induced cell death and increased Bcl-2 and procaspase-3 expression, whereas the expression of Bax and cleaved PARP were decreased in a dose-dependent manner compared to the control. Furthermore, Hwabuk 94 inhibited the t-BHP-induced p38 MAP kinase and extracellular signal-regulated kinase 1/2, but not c-Jun N-terminal kinase activations. Finally, these findings suggest that Hwabuk 94 kiwifruit might attenuate t-BHP-induced PC12 cell cytotoxicity, at least in part, through the inhibition of signaling pathways mediated by the ERK1/2 and p38 MAP kinase.

• 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.

• The Journal of Internal Korean Medicine
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• v.26 no.1
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• pp.107-118
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• 2005

Objectives : Peroxynitrite $(ONOO^-)$, fonned from the reaction of $O_2^-$ and NO, is a cytotoxic species that can oxidize several cellular components such as proteins, lipids and DNA. It has been implicated in the aging process and age-related disease such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. Due to the lack of endogenous enzymes to thwart $ONOO^-$ activation, developing a specific $ONOO^-$ scavenger is remarkably important. The aim of this study was to investigate scavenging activities of $ONOO^-$ and its precursors, NO and $O_2^-$ and its scavenging mechanism of Ojawhan. Methods : To investigate scavenging activities of $ONOO^-$, NO, $O_2^-$ and its scavenging mechanism using fluorescent probes, DCFDA, DAF-2 and DHR 123. The $ONOO^-$ scavenging activity on Ojawhan was assayed by measuring oxidized dihydrorhodamine 123 (DHR 123) by fluorometry. Oxidative stress was induced by strong oxidants t-butyl hydroperoxide (t-BHP). Endothelial cell (YPEN-1) was used for detection of intracellular oxidative stress. Results : Ojawhan markedly scavenged authentic $ONOO^-$, $O_2^-$ and NO. It also inhibited $ONOO^-$ induced by $O_2^-$ and NO which are derived from SIN-1. Furthennore, ${\underline{Ojawhan}}$ blocked lipopolysaccharide (LPS)-induced $ONOO^-$, $O_2^-$ and NO generation utilizing kidney homogenates of LPS-injected mouse and inhibited t-BHP-induced ROS and $ONOO^-$ in endothelial cell culture system. Conclusions : These results suggest that Ojawhan be developed as an effective $ONOO^-$ scavenger for the prevention of $ONOO^-$ involved diseases and age-related diseases.

• The Journal of Internal Korean Medicine
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• v.28 no.1
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• pp.80-91
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• 2007

Objectives : Peroxynitrite (ONOO-), superoxide anion radical (?O2-) and nitric oxide (NO) are cytotoxic because they can oxidize several cellular components such as proteins, lipids and DNA. They have been implicated in the aging process, and age-related diseases such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. The aim of this study was to investigate Sabohwan's activity for scavenging ONOO- and its precursors. NO and ?02-. Methods : For this study, the fluorescent probes, namely 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), 4.5-diaminofluorescein (DAF-2) and dihydrorhodamine 123 (DHR 123) were used. Results : Sabohwanblocked tert-butylhydroperoxide (t-BHP)-induced cell death in a dose-dependent fashion. It scavenged t-BHP-induced ONOO-, NO and ?O2- in YPEN cells. Sabohwan inhibited the generation of ONOO-, NO and ?O2- in the lipopolysaccharide (LPS)-treated mouse kidney postmitochondria both in vitro and in vivo. The lipid peroxide level increased and glutathione level decreased in the LPS-treated mice, whereas the ones in the Sabohwanadministered group among the LPS-treated mice reversed toward their natural levels. Conclusions : These results suggest that Sabohwanis an effective ONOO-, ?O2- and NO scavenger, and thereby it might have a potential role as a therapy against the aging process and age-related diseases.