• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.143.1-143.1
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• 2003

In this study. we investigated whether the cardioprotective effect shown by quercetin 3-O-$\alpha$-arabinofuranoside extracted from Lindera erythrocarpa against ROS-induced cell death in H9c2 cardiac myocytes. Cell death was induced by BSO, buthionine sulfoximine, which inhibits GSH level and subsequntly increase ROS level. Cell death was quntitatively determined by measuring lactate dehydrogenase (LDH) activity. (omitted)

• Asian Pacific Journal of Cancer Prevention
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• 제16권8호
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• pp.3213-3222
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• 2015

Background: Cancer metastasis depends on cell motility which is driven by cycles of actin polymerization and depolymerization. Reactive oxygen species (ROS) and metabolic oxidative stress have long been associated with cancer. ROS play a vital role in regulating actin dynamics that are sensitive to oxidative modification. The current work aimed at studying the effects of sub-lethal metabolic oxidative stress on actin cytoskeleton, focal adhesion and cell migration. Materials and Methods: T47D human breast cancer cells were treated with 2-deoxy-D-glucose (2DG), L-buthionine sulfoximine (BSO), or doxorubicin (DOX), individually or in combination, and changes in intracellular total glutathione and malondialdehyde (MDA) levels were measured. The expression of three major antioxidant enzymes was studied by immunoblotting, and cells were stained with fluorescent-phalloidin to evaluate changes in F-actin organization. In addition, cell adhesion and degradation ability were measured. Cell migration was studied using wound healing and transwell migration assays. Results: Our results show that treating T47D human breast cancer cells with drug combinations (2DG/BSO, 2DG/DOX, or BSO/DOX) decreased intracellular total glutathione and increased oxidized glutathione, lipid peroxidation, and cytotoxicity. In addition, the drug combinations caused a reduction in cell area and mitotic index, prophase arrest and a decreased ability to form invadopodia. The formation of F-actin aggregates was increased in treated T47D cells. Moreover, combination therapy reduced cell adhesion and the rate of cell migration. Conclusions: Our results suggest that exposure of T47D breast cancer cells to combination therapy reduces cell migration via effects on metabolic oxidative stress.

• Journal of Preventive Medicine and Public Health
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• 제35권4호
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• pp.269-274
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• 2002

Balb/c 마우스의 복강내에 Abelson leukemia virus (A-MuLV)를 주입하여 발생시킨 대식세포주 RAW 264.7 세포의 배양조건에 납과 NAC및 BSO를 첨가하여 세포생존율과 NO 및 ATP 생성량의 변화를 관찰한 결과, 납을 처리한 기본배양조건에서 RAW 264.7 세포의 생존율은 각 농도에서 차이가 없었으며 NO의 생성량은 $0.5{\mu}M$의 납농도에서부터 용량 의존적으로 감소하였으나 ATP의 생성량은 각 농도군에서 차이가 없었다. NAC을 전처리하고 납을 처리한 배양조건에서의 NO 및 ATP의 생성량은 대조군과 차이가 없었다. BSO를 전처리하고 납을 처리한 배양조건에서의 NO의 생성량은 납만 처리했을 때와 달리 각각의 농도군에서 대조군과 차이가 있었다. ATP생성량은 역시 차이가 없었다. 이상의 결과에서 납의 농도가 증가함에 따라 ATP의 생성량은 변화가 없으면서 NO가 감소하는 것을 볼 때, 납에 의한 대식세포에서 NO생성의 억제기전은 수은 및 카드뮴 등과 같이 미토콘드리아에 영향을 미쳐 ATP생성이 억제됨으로 L-arginine-NO경로에서 ATP를 필요로 하는 iNOS가 작용을 못하여 NO 생성이 저하되는 기전과는 다른 기전이 있음을 보여준다. 또한 iNOS의 조효소인 세포내 GSH를 증가시키는 NAC을 전처리했을 때 NO의 생성량이 대조군 수준으로 회복되고 세포내 GSH를 감소시키는 BSO를 전처리했을 때는 오히려 NO의 생성량에 영향을 미치지 않는 납의 농도에서조차 NO 생성의 감소가 일어난 것으로 볼 때 GSH는 대식세포에서 NO생성을 저하시키는 납의 독성에 보호작용이 있음을 확인할 수 있었다.

• Archives of Pharmacal Research
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• 제27권4호
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• pp.454-459
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• 2004

The present study investigated the effects of gossypin, 3,3',4',5,7,8-hexahydroxyflavone 8-glucoside, on the toxicity induced by oxidative stress or $\beta$-amyloid ($A_{\beta}$) in primary cultured rat cortical cells. The antioxidant properties of gossypin were also evaluated by cell-free assays. Gossypin was found to inhibit the oxidative neuronal damage induced by xanthinelxanthine oxidase or by a glutathione depleting agent, D,L-buthionine (S,R)-sulfoximine. In addition, gossypin significantly attenuated the neurotoxicity induced by $A_{{\beta}(25-35)}$. Furthermore, gossypin dramatically inhibited lipid peroxidation initiated by $Fe^{2+}$ and ascorbic acid in rat brain homogenates. It also exhibited potent radical scavenging activity generated from 1 ,1-diphenyl-2-picrylhydrazyl. These results indicate that gossypin exerts neuroprotective effects in the cultured cortical cells by inhibiting oxidative stress- and $A_{\beta}$-induced toxicity, and that the antioxidant properties of gossypin may contribute to its neuroprotective actions.

• BMB Reports
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• 제32권2호
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• pp.161-167
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• 1999

The role of oxidative stress in the initiation and the complication of diabetes was examined by monitoring blood glucose increase and oxidative DNA damage in rats treated with alloxan or streptozotocin (STZ). Oxidative DNA damage was assessed by quantitating 8-oxo-2'-deoxyguanosine ($oxo^8dG)$ excreted in urine and the $oxo^8dG$ accumulated in pancreas DNA. Both alloxan and STZ treatments resulted in an abrupt increase in blood glucose and significant increases in urinary and pancreatic $oxo^8dG$. Pretreatment of buthionine sulfoximine (BSO), a glutathione-depleting agent, slightly potentiated the increase of blood glucose and urinary $oxo^8dG$ in the alloxan- and STZ-treated rats. Furthermore, the BSO pretreatment caused significant amplification of pancreatic $oxo^8dG$ increase in the rats. On the other hand, pretreatment with 1,10- phenanthroline (o-phen), a chelator of divalent cations, showed different results between alloxan- and STZ-treated rats. The o-phen pretreatment completely blocked diabetes and the increase of $oxo^8dG$ by alloxan treatment, while it potentiated the increase of blood glucose and $oxo^8dG$ by STZ treatment. The results demonstrate that the causative effect of alloxan on diabetes may be the generation of reactive oxygen species through a Fenton type reaction, but that of STZ may not.

• Molecules and Cells
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• 제19권1호
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• pp.131-136
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• 2005

The ETV6-NTRK3 gene fusion, first identified in the chromosomal translocation in congenital fibrosarcoma, encodes a chimeric protein tyrosine kinase with potent transforming activity. ETV6-NTRK3-dependent transformation involves the joint action of NTRK3 signaling pathways, and aberrant cell cycle progression resulting from activation of Mek1 and Akt. The level of glutathione (GSH) was found to be markedly increased in ETV6-NTRK3-transformed NIH3T3 cells. The activities of the two GSH biosynthetic enzymes as well as of glutathione peroxidase, together with their mRNAs, were also higher in the transformed cells. The transformed cells were able to grow in the presence of GSH-depleting agents, whereas the control cells were not. L-Buthionine-(S,R)-sulfoximine (BSO) inhibited activation of Mek1 and Akt in the transformed NIH3T3 cells. These observations imply that up-regulation of GSH biosynthesis plays a central role in ETV6-NTRK3-induced transformation.

• The Korean Journal of Physiology and Pharmacology
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• 제5권3호
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• pp.271-278
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• 2001

Oxidative stress and methylglyoxal (MG), a reactive dicarbonyl metabolites produced by enzymatic and non-enzymatic reaction of normal metabolism, induced aldose reductase (AR) expression in rat aortic smooth muscle cells (SMC). AR expression was induced in a time-dependent manner and reached at a maximum of 4.5-fold in 12 h of MG treatment. This effect of MG was completely abolished by cyclohemide and actinomycin D treatment suggesting AR was synthesized by de novo pathway. Pretreatment of the SMC with N-acetyl-L-cysteine significantly down-regulated the MG-induced AR mRNA. Furthermore, DL-Buthionine-(S,R)-sulfoximine, a reagent which depletes intracellular glutathione levels, increased the levels of MG-induced AR mRNA. These results indicated that MG induces AR mRNA by increasing the intracellular peroxide levels. Aminoguanidine, a scanvenger of dicarbonyl, significantly down-regulated the MG-induced AR mRNA. In addition, the inhibition of AR activities with statil, an AR inhibitor, enhanced the cytotoxic effect of MG on SMC under normal glucose, suggesting a protective role of AR against MG-induced cell damages. These results imply that the induction of AR by MG may contribute to an important cellular detoxification of reactive aldehyde compounds generated under oxidative stress in extrahepatic tissues.

• 한국식품영양과학회지
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• 제46권2호
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• pp.161-168
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• 2017

본 연구에서는 산화적 스트레스로 유도한 시신경 세포사멸에 대한 차조기 물 추출물(PFE)의 효과를 확인하였다. Staurosporine으로 분화된 ssdRGC-5 세포에 buthionine과 glutamate(B/G)로 산화적 스트레스를 유도하였으며, LDH release assay, MTT reduction assay를 통하여 PFE가 농도 의존적으로 B/G에 의한 세포사멸을 억제함을 관찰하였다. 세포사멸의 기전을 연구하기 위해 caspase 활성, 세포 내 ROS 생성량, 세포고사 관련 단백질 발현을 관찰한 결과, B/G에 의해 증가한 ROS 생성량, caspase 활성을 PFE가 억제하였고, 세포질로 방출된 cytochrome c와 미토콘드리아로 이동한 Bax도 감소함을 확인하였다. 이상의 결과로부터 차조기는 산화적 스트레스로 유도된 시신경 세포사멸 과정에서 항산화 효과와 미토콘드리아성 세포사멸을 완화함으로써 세포 보호 작용을 나타냄을 확인하였다.

• BMB Reports
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• 제42권9호
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• pp.561-567
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• 2009

Although many plant-derived phenolic compounds display antioxidant effects in biological systems, their mechanism of action remains controversial. In this study, the mechanism by which p-coumaric acid (p-CA) performs its antioxidant action was investigated in bovine aortic endothelial cells under oxidative stress due to high levels of glucose (HG) and arachidonic acid (AA), a free fatty acid. p-CA prevented lipid peroxidation and cell death due to HG+AA without affecting the production of reactive oxygen species. The antioxidant effect of p-CA was not decreased by buthionine-(S,R)-sulfoximine, an inhibitor of cellular GSH synthesis. In contrast, pretreatment with p-CA caused the induction of peroxidases that decomposed t-butyl hydroperoxide in a p-CA-dependent manner. Furthermore, the antioxidant effect of p-CA was significantly mitigated by methimazole, which was shown to inhibit the catalytic activity of 'p-CA peroxidases' in vitro. Therefore, it is suggested that the induction of these previously unidentified 'p-CA peroxidases' is responsible for the antioxidant effect of p-CA.

• Toxicological Research
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• 제16권1호
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• pp.1-8
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• 2000

Astrocytes generate free radicals including nitric oxide (NO) and reactive oxygen intermediates(ROI) which in turn play roles in the pathogenesis of degenerative diseases and sclerotic changes of the brain. This study was designed to evaluate the mechanism that free radicals contribute to the cytotoxicty of rat neonatal primary astrocytes. Treatment with NO donors alone including soldium nitroprusside(SNP), S-nitrosoglucathinoe (GSNO), and S-nitroso-n-acetylpenicillamine (SNAP) showed a little effect on the death of rat neonatal primary astrocytes, whereas SNP markedly induced the death of RAW 264.7 cells. ROI inculding H2O2 and O2 donor also slightly induced the death of rat primary astrocytes. However, 3-morpholinosydnonimine(SIN-1), a donor of peroxynitrite (ONOO), which is a reactive compound of NO with superoxide, significantly decreased the viability of rat primary astrocytes in a dose-dependent manner. Cells were retarded in outgrowth of viability of cellular processes with cell shrinkage and detachment from culture dishes. Hoechst staining demonstrated that SIN-1-induced cell death might be due to an apoptosis which was characterized by nuclear condensation and fragmentation. SIN-1-induced apoptosis was prevented by the pretreatment with superoxide dismutase (SOD) and catalase in rat primary astorocytes. Furthermore, prevention of the generation of reduced glutathione (GSH) by DL-buthionine-[S, R]-sulfoximine (BSO) aggravated the cytotoxic effects of SNP, benzene triol, and SIN-1 in rat primary astrocytes. Taken together, it is suggested that peroxynitrite may be a major effector of apoptosis and cellular antioxidant system is important for cell survival in rat prima교 astrocytes.