• 대한한의학회지
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• 제23권4호
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• pp.55-63
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• 2002

Objectives: Peroxynitrite ($ONOO^{-}$), formed from the reaction of superoxide <${\cdot}O_2^{-}$) and nitric oxide (NO), is a cytotoxic species that can oxidize several cellular components such as proteins, lipids and DNA. It has been implicated in diseases such as aging process, Alzheimer's disease, rheumatoid arthritis, cancer and arteriosclerosis. Due to the lack of endogenous enzymes responsible for $ONOO^{-}$ inactivation, developing a specific $ONOO^{-}$ scavenger is of considerable importance. The aim of this study was to evaluate $ONOO^{-}$ scavenging activity and its mechanism in Cheonga-hwan (CAH). Methods: The $ONOO^{-}$ scavenging activity in CAH was assayed by measuring oxidized dihydrorhodamine 123 (DHR 123) by fluorescence. The scavenging efficacy was expressed as $IC_{50}$, showing the concentration of each sample required to cause 50% inhibition of DHR 123 oxidation. In a separate study, the protective effect of CAR on $ONOO^{-}$-induced nitration of bovine serum albumin (BSA) was investigated using immunoassay with a monoclonal anti-nitrotyrosine antibody, and a horseradish peroxidase-conjugated anti-mouse secondary antibody from sheep. Results: CAH showed potent scavenging activities of $ONOO^{-}$, NO and ${\cdot}O_2^{-}$. The data demonstrated that CAH led to decreased $ONOO^{-}$-mediated nitration of tyrosine through electron donation. CAH showed significant inhibition on nitration of bovine serum albumin by $ONOO^{-}$ in a dose-dependent manner. Conclusions: CAH can be developed as an effective peroxynitrite scavenger for the prevention of the $ONOO^{-}$ involved diseases.

• 대한한방내과학회지
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• 제26권1호
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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.

• 대한한방내과학회지
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• 제24권1호
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• pp.75-83
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• 2003

Objectives : Peroxynitrite($ONOO^-$), formed from the reaction of $O2^-$ and NO, is a cytotoxic species that can oxidize several cellular components such as proteins, lipids and DNA. It has been involved 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 scavenging activities of $ONOO^-$ and its precursors, NO and $O_{2^-}$ and its scavenging mechanism of Zingiberis Rhizoma (ZR). Methods : To investigate scavenging activities of $ONOO^-,\;NO,\;O_{2^-}$ and its scavenging mechanism, we used fluorescent probes like DCFDA, DAF-2 and DHR 123. The $ONOO^-$ scavenging activity on ZR was assayed by measuring oxidized dihydrorhodamine 123 (DHR 123) by fluorometry. The scavenging efficacy was expressed as IC50, showing the concentration of each sample that is required to cause 50% inhibition of DHR 123 oxidation. In a separate study, the protective effect of ZR on $ONOO^-$-induced nitration of bovine serum albumin was investigated through immuno-assay with a monoclonal anti-nitryrosine antibody, and a horseradish peroxidase-conjugated anti-mouse secondary antibody from sheep. Results : ZR markedly scavenged authentic $ONOO^-,\;O_{2^-}$ and NO. It also inhibited $ONOO^-$ induced by $O_{2^-}$ and NO which are derived from SIN-1. The data demonstrated that ZR led to decreased $ONOO^-$ mediated nitration of tyrosine through electron donation. It also inhibited the nitration of bovine serum albumin induced by $ONOO^-$ in a dose-dependent manner. Furtheremore, it blocked LPS-induced ROS and RNS generation. Conclusions : These results suggest that ZR can be developed as an effective $ONOO^-$ scavenger for the prevention of aging process and age-related diseases.

• 대한한방내과학회지
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• 제27권1호
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• pp.178-187
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• 2006

Objectives : Peroxynitrite $(ONOO^-),\;{\cdot}O_2^-$ and nitric oxide (NO) are 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. The aim of this study was to investigate $ONOO^-$ scavengactivities, and that of its precursors, NO and ${\cdot}O_2^-$ of Samjunghwan. Methods : To investigate $ONOO^-,\;NO,\;{\cdot}O_2^-$ scavenging activities, fluorescent probes, namely 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) and dihydrorhodamine 123 (DHR 123) were used. Results : Samjunghwan markedly scavenged authentic $ONOO^-,\;{\cdot}O_2^-$ and NO. It also inhibited $ONOO^-$ induced by .02' and NO which are derived from SIN-I. Furthermore, Samjunghwan inhibited $ONOO^-$, ${\cdot}O_2^-$ and NO generation in LPS-treated ICR mouse kidney postmitochondria. Conclusions : These results suggest that Samjunghwan is an effective $ONOO^-,\;{\cdot}O_2^-$ and NO scavenger, and that this substance has a potential role as an inhibitor of aging process, and in therapy against age-related diseases.

• 한국자원식물학회지
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• 제21권4호
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• pp.254-259
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• 2008

약용식물 9종의 물 추출물에 대한 항산화 효과를 탐색하기 위해 in vitro 조건에서 강력한 산화력과 독성을 갖는 $ONOO^-$와 ${\cdot}$OH radical의 소거효과를 측정하였다. 그 결과, 감잎, 구기자, 꾸지뽕나무, 담쟁이덩굴, 마디풀, 비파나무, 으름덩굴, 참취 추출물은 10${\mu}g/ml$ 농도에서 50% 이상의$ONOO^$ 소거효과를 나타냈다. 이러한 $ONOO^-$ 소거효과는 양성 대조물인 penicillamine(94.08${\pm}$3.04)에 비해 낮았지만 비파나무 물 추출물(89.87${\pm}$4.57)이 다른 시료에 비해 가장 강력한 $ONOO^-$ 소거효과를 나타냈다. 또한, 가는잎쐐기풀과 감잎 물 추출물은 1mg/ml 농도에서 양성대조물인 thiourea 보다 효과적인${\cdot}$OH radical 억제효 과를 나타냈다. 결론적으로 9종의 약용식물은 $ONOO^-$와${\cdot}$OH radical과 연관된 산화적 스트레스에 의한 세포와 조직의 손상을 억제시킬 수 있는 천연 항산화제로 밝혀졌다.

• 대한한방내과학회지
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• 제25권2호
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• pp.268-276
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• 2004

Objective : Peroxynitrite $(ONOO^-)$, formed 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. The aim of this study was to investigate scavenging activities of $ONOO^-$ and its precursors. NO and $O_2^-$ and its scavenging mechanism using fluorescent probes, DCFDA, DAF-2 and DHR 123.. Methods : Psoralea corylifolia was crushed. The crushed Psoralea corylifolia was extracted 3 times, each time with 3 volumes of methyl alcohol at $60^{\circ}C$ for 24 h. The extract was filtered and evaporated under reduced pressured using a rotary evaporator to yield 16g. This was done to investigate scavenging activities of $ONOO^-$, NO, $O_2^-$ and its scavenging mechanism using fluorescent probes, DCFDA, DAF-2 and DHR 123. Results : After Psoralea corylifolia was added authentic $ONOO^-,\;{\cdot}\;O_2^-$ and NO was markedly scavenged. Also, $ONOO^-$ induced by $O_2^-$ and NO (these derived from SIN-1) was inhibited. The data showed a decrease in $ONOO^-$ mediated nitration of tyrosine through electron donation after Psoralea corylifolia was added. Data showed a dose-dependent correlation with inhibition of nitration of bovine serum albumin induced by $ONOO^-$, Furtheremore, LPS-induced ROS and RNS generation was blocked. Conclusions: These results suggest potential for use of Psoralea corylifolia as an effective $ONOO^-$ scavenger to counter the aging process and age-related diseases.

• 대한한의학회지
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• 제27권2호
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• pp.171-181
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• 2006

Objectives : Peroxynitrite($ONOO^-$), superoxide anion(${\cdot}{O_2}^-$) and nitric oxide (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. The aim of this study was to investigate scavenging activities for $ONOO^-$ and its precursors, NO and ${\cdot}{O_2}^-$ of Vespae Nidus. Methods : Dichlorodihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) and dihydrorhodamine 123 (DHR 123) were used to investigate scavenging activities of $ONOO^-,\;NO,\;{\cdot}{O_2}^-$. Six-months-old ICR mice were used. After mice were injected with lipopolysaccharides(LPS), kidney organization was evaluated. Three comparison groups of ICR mice were used : a normal group, an experimental group that was fed Vespae Nidus extract and then injected with LPS, and a control group that was injected with LPS. Scavenging activities of $ONOO^-,\;NO,\;{\cdot}{O_2}^-$ in these groups were measured in the same way. Results : Vespae Nidus markedly scavenged authentic $ONOO^-,\;{\cdot}{O_2}^-$ and NO. It also inhibited $ONOO^-$ induced by ${\cdot}{O_2}^-$ and NO which are derived trom SIN-1. Furthermore, it inhibited $ONOO^-,\;{\cdot}{O_2}^-$, and NO generation by Vespae Nidus in LPS-treated ICR mouse kidney postmitochondria. Conclusions : These results suggest that Vespae Nidus might be developed as an effective $ONOO^-,\;{\cdot}{O_2}^-$, and NO scavenger for the prevention of the aging process and age-related diseases.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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• pp.126-126
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• 2001

Peroxynitrite (ONOO$^{-}$) is one of cytotoxic species produced by the reaction between superoxide(ㆍ $O_2$$^{-}$) and nitric oxide (NO). The aim of this study was to characterize ONOO$^{-}$ scavenging constituents from herbs. Methanolic extract derived from one hundred fifty nine herbs were screened out for their ONOO$^{-}$ scavenging activities. It was investigated that about 33 herbs was excellent scavengers of ONOO$^{-}$. The extracts exhibited dose-dependent ONOO$^{-}$ scavenging activities.

• 동의생리병리학회지
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• 제21권6호
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• pp.1499-1505
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• 2007

Peroxynitrite ($ONOO^-$) is a reactive oxidant formed from superoxide anion radical (${\cdot}\;O_2-$) and nitric oxide (NO), which can oxidize cellular components such as essential protein, non-protein thiols, DNA, low-density lipoproteins and membrane phospholipids. ${\cdot}\;O_2-$ and $ONOO^-$ have contributed to the pathogenesis of diseases such as stroke, heart disease, Alzheimer's disease and atherosclerosis. Because of damaging effects of ${\cdot}\;O_2-$ and $ONOO^-$ oxidants, Vespae Nidus, which has been known to strengthen the kidneys to preserve the vital energy. was tested as a potential specific scavenger of those oxidants. In this study, the viability of Vespae Nidus (1, 10, 50 g/ml) to scavenge ${\cdot}\;O_2-$, NO, $ONOO^-$ and so to protect cells against tert-butylhydroxyperoxide (t-BHP) induced cell death was tested. The levels of ${\cdot}\;O_2-$ and $ONOO^-$ were detected by staining with DCFH-DA and DHR 123, respectively. Protein expression levels of COX-2, iNOS and $NF{-\kappa}B$ were assayed by western blot. Vespae Nidus blocked t-BHP-induced cell death in a dose-dependent fashion. Vespae Nidus inhibited t-BHP-induced production of ${\cdot}\;O_2-$, NO and $ONOO^-$ in YPEN cells. The lipid peroxide level was increased and glutathione level was decreased in lipopolysaccharide (LPS)-treated ICR mouse, whereas the ones in the Vespae Nidus-administered group were regulated beneficially. Vespae Nidus inhibited the expression of COX-2, iNOS and NF-κB (p65 and p50) genes in LPS-treated ICR mouse. The present study suggests that Vespae Nidus is a powerful antioxidant and promotes cellular defense activity by scavenging the toxic oxidants such as ${\cdot}\;O_2-$ and $ONOO^-$.

• Journal of Ginseng Research
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• 제34권3호
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• pp.205-211
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• 2010

Korean red ginseng (KRG) has been used worldwide as a traditional medicine for the treatment of various diseases, including cancer. In this study, we determined the effect of KRG on the responses of HaCaT cells to peroxynitrite ($ONOO^-$). Cells has been used worldwide as a traditional medicine for the treatment of various diseases, including cancer. In this study, we determined the effect of KRG on the responses of HaCaT cells to peroxynitrite ($ONOO^-$). Cells treated with $ONOO^-$ (2 mM) prior to incubation with control medium for 12 hours displayed reduced viability, as determined using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay (viability about 48% of that of non-treated control cells). When KRG was added to the post-incubation medium, the negative effects of $ONOO^-$ on cell viability were significantly reduced. Reverse transcription-polymerase chain reaction analysis indicated that KRG alone did not significantly alter p53 or "growth arrest and DNA damage" (GADD)45 mRNA levels. However, the addition of KRG to the post-incubation medium significantly and dose-dependently reduced levels of p53 and GADD45 mRNA in $ONOO^-$-treated cells. Western blot analyses revealed that incubation with KRG decreased p53 and GADD45 protein levels in $ONOO^-$-treated cells, relative to those in cells incubated with control medium. Collectively, these results suggest that Korean red ginseng extract protects cells against $ONOO^-$-induced genotoxicity by increasing cell viability through modulating the expression of p53 signaling intermediates.