• 한국패류학회지
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• 제29권1호
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• pp.15-21
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• 2013

일산화질소 (NO) 는 면역계에서 세포내 외의 신호전달에 관여하는 물질로 생물의 생리적, 병리학적 기작을 조절한다. 본 연구는 바지락 혈구의 NO 농도 측정을 위해 4,5-diaminofluorescein diacetate (DAF-2 DA) 를 이용한 DAF assay의 적용이 가능한지 확인하고자 화상분석법, 형광흡광도 측정법 및 유세포분석 기법 등을 이용하였다. 연구결과 인위적인 바지락 혈구의 NO 생성을 위해 L-arginine을 첨가한 경우 대조구에 비하여 NO 생성이 유의적으로 증가하였고, 반대로 NO 저해제인 L-NAME를 첨가한 경우 NO 생성은 급격히 감소하였다. 이러한 결과는 본 조사에 이용된 화상분석법, 형광흡광도 측정법 및 유세포분석 기법 등 모든 조사 방법에서 동일하게 확인되었다. 특히 3가지 측정 방법 중 유세포 분석법은 측정의 신속성, 신뢰성 및 정확성을 담보할 수 있는 유용한 방법으로 판단된다. 따라서 유세포 분석기를 이용한 NO 측정은 향후 바지락의 생리적 병리적 특성을 확인하는데 유용한 마커로써 이용될 수 있을 것으로 기대된다.

• 대한한방내과학회지
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• 제29권2호
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• pp.421-431
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• 2008

Objectives : This study was to investigate the inhibitory effects of Ulmus davidiana on the generation of peroxynitrite $(ONOO^{-})$, nitric oxide (NO) and superoxide anion radicals $(O_{2}^{-})$ in the endothelial cells of rat vessels. The effects of Ulmus davidiana on the expression of inflammation-related proteins, $NF-{\kappa}B$ (p50, p65), COX-2, and iNOS, were examined by western blotting. Methods : For this study, fluorescent probes, namely dihydrorhodamine 123 (DHR 123), 4,5-diaminofluorescein (DAF-2) and 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) were used. Western blotting was performed via using anti-$NF-{\kappa}B$ (p50, p65), anti-COX-2, and anti-iNOS, respectively. Results : Ulmus davidiana inhibited the generation of $ONOO^{-}$, NO and $(O_{2}^{-})$ in the lipopolysaccharide (LPS)-treated endothelial cells of rat vessels in vitro. Ulmus davidiana inhibited the expression of COX-2 and iNOS genes by means of decreasing the $NF-{\kappa}B$ activation. Conclusions : These results suggest Ulmus davidiana is effective on inhibiting the generation of $ONOO^{-}$, NO and $O_{2}^{-}$, and that therefore it might have a potential role as a treatment for the inflammatory process and inflammation-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.

• 대한한방내과학회지
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• 제28권1호
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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.

• 대한한방내과학회지
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• 제30권1호
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• pp.36-50
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• 2009

Objectives : This study was aimed to verify the cytoprotective function, antioxidative effect and inflammation genes inhibitory effects of Lycium chinense Milie. Therefore the generation of superoxide anion radical ( $O_2\;^-$), peroxynitrite ($ONOO^-$), nitric oxide (NO) and prostaglandin $E_2$ $(PGE_2)$ was investigated in the renal epithelial cells of mouse. Effects of Lycium chinense Milie on the expression of inflammation-related proteins, $IKK-{\alpha}$. $p-IKK-\alpha\beta$, $p-I{\kappa}B-\alpha$, $NF-{\kappa}B$ (p50, p65), COX-2 and iNOS, were examined by western blotting. Methods : For this study, the fluorescent probes were used, namely dihydrorhodamine 123 (DHR 123), 4.5-diaminofluorescein (DAF-2) and 2',7'-dichlorodihydrofluorescein diacetate (DCFDA). Western blotting was performed using anti-$IKK-\alpha$, anti-phospho $IKK-\alpha\beta$, anti-phospho $I{\kappa}B-\alpha$, anti-$NF-{\kappa}B$ (p50, p65), anti-COX-2 and anti-iNOS, respectively. Results : Lyciutn chinense Milie reduced $H_{2}O_{2}$-induced cell death dose-dependently. It inhibited the generation of $O_2\;^-$, $ONOO^-$, NO and $PGE_2$ in the $H_{2}O_{2}$-treated renal epithelial cells of mouse in vitro. Lycium chinense Milie inhibited the expression of $IKK-\alpha$, $p-IKK-\alpha\beta,\;p-I{\kappa}B-\alpha$, COX-2 and iNOS genes by means of decreasing activation of $NF-{\kappa}B$. Conclusions : According to above results. Lycium chinense Milie recommended to be applied in treatment for the inflammatory process and inflammation-related diseases.

• 동의생리병리학회지
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• 제26권4호
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• pp.483-490
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• 2012

This study was designed to identify the effects of Polygoni cuspidati Radix(PCR) on the generation of superoxide anion radicals (${\cdot}O_2{^-}$), nitric oxide (NO), peroxynitrite ($ONOO^-$) in the renal epithelial cells of mouse(LLC-$PK_1$). The effects of PCR on the expression of inflammation-related proteins, IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$, VCAM-1, were examined by western blotting. For this study, the fluorescent probes, namely dihydrorhodamine 123 (DHR 123), 2',7'-dichloro dihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) were used. Protein expression levels of IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$, VCAM-1 were assayed by western blot. PCR reduced $H_2O_2$-induced cell death dose-dependently. It inhibited the generation of ${\cdot}O_2{^-}$, NO, $ONOO^-$ and $PGE^2$ in the $H_2O_2$-treated LLC-PK1 cells in vitro. PCR inhibited the espression of IKK-${\alpha}$, phospho-$I{\kappa}B-{\alpha}$, COX-2, iNOS, IL-$1{\beta}$ and VCAM-1 genes by means of decreasing the NF-${\kappa}B$ activation. These results suggest that PCR is an effective NO, ${\cdot}O_2{^-}$, $ONOO^-$ scavenger, and this substance recommended to be applied in treatment for the inflammatory process and inflammation-related disease.

• 대한한방내과학회지
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• 제27권3호
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• pp.688-697
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• 2006

Objectives : Peroxynitrite ($ONOO^-$), $O_2^-$ 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 $ONOO^-$ scavenging activities. and that of its precursors. NO and $O_2^-$ of Trigonellae Semen. Methods : To investigate $ONOO^-$. NO. $O_2^-$ scavenging activities, fluorescent probes, namely 2'.7'-dichlorodihydrofluorescein diacetate (DCFDA). 4.5-diaminofluorescein (DAF-2) and dihydrorhodamine 123 (DHR 123) were used. Protein expression levels of iNOS, COX-2, NF-${\kappa}B$, and VCAM-1 were assayed by western blot. Results : Trigonellae Semen markedly scavenged authentic $ONOO^-$, $O_2^-$ and NO. It also inhibited $ONOO^-$ induced by $O_2^-$ and NO which are derived from SIN-1. Furthermore. Trigonellae Semen inhibited $ONOO^-$, $O_2^-$ and NO generation in LPS-treated ICR mouse kidney postmitochondria. Trigonellae Semen inhibited gene expression of iNOS, COX-2, VCAM-l and NF-${\kappa}B$ (p65) activation. Conclusions : These results suggest that Trigonellae Semen is an effective $ONOO^-$, $O_2^-$ and NO scavenger. and that this substance has a potential role as an inhibitor of the aging process, and in therapy against age-related diseases.

• 대한한의학회지
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• 제29권1호
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• pp.106-116
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• 2008

Objectives : Peroxynitrite $(ONOO^-)$, superoxide anion radical $({\cdot}O_2^-)$ 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 processes, and age-related diseases such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. The aim of this study was to investigate the $ONOO^-$, NO, and $({\cdot}O_2^-)$ scavenging and anti-inflammatory activities of Mori Fructus in ob/ob mice. Methods : Mice were grouped and treated for 5 weeks as follows. Both the normal lean (C57/BL6J black mice) and control obese (ob/ob mice) groups received the standard chow. The experimental groups were fed a diet of chow supplemented with 7.5, 15 and 30 mg Mori Fructus per 1 kg of body weight for 14 days. For this study, the fluorescent probes, namely 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) and dihydrorhodamine 123 (DHR 123) were used. Western blotting was performed using anti-phospho $I{\kappa}B-\alpha$, anti-IKK-$\alpha$, anti-NF-${\kappa}B$ (p50, p65), anti-COX-2 and anti-iNOS respectively. Results : Mori Fructus inhibited the generation of $ONOO^-$, NO and $({\cdot}O_2^-)$ in the lipopolysaccharide (LPS)-treated mouse kidney postmitochondria in vitro. The generation of $ONOO^-$, NO and $({\cdot}O2^-)$ were inhibited in the Mori Fructus-administered ob/ob mice groups. The GSH/GSSG ratio decreased in the ob/ob mice, whereas they improved in the Mori Fructus-administered groups. Mori Fructus inhibited the expression of phospho $I{\kappa}B-\alpha$, IKK-$\alpha$, COX-2, iNOS genes, and thereby the activation of NF-$I{\kappa}B$. Conclusions : These results suggest that Mori Fructus is an effective $ONOO^-$, $({\cdot}O_2^-)$ and NO scavenger, and therefore it might be a potential therapeutic drug against the inflammation process and inflammation-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.

• 동의생리병리학회지
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• 제22권6호
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• pp.1475-1481
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• 2008

This study was to investigate the inhibitory effects of Mori Fructus on the generation of peroxynitrite ($ONOO^-$), nitric oxide (NO) and superoxide anion radical (${\cdot}O_2{^-}$) in the endothelial cells of rat vessels. The aim of this study was to investigate the $ONOO^-$, NO, ${\cdot}O_2{^-}$ scavenging and anti-inflammatory activitives of Mori Fructus. For this study, the fluorescent probes, namely dihydrorhodamine 123 (DHR 123), 4,5-diaminofluorescein (DAF-2) and 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) were used. Western blotting was performed using anti-NF-${\kappa}B$ (p50, p65), anti-COX-2, anti-iNOS antibodies, respectively. Mori Fructus prevented lipopolysaccharide (LPS)-induced cell death in YPEN cells. Mori Fructus inhibited the generation of $ONOO^-$, NO and ${\cdot}O_2{^-}$ in the LPS-treated cells. Mori Fructus inhibited the expression of COX-2 and iNOS genes by means of decreasing the NF-${\kappa}B$ activation. These results suggest Mori Fructus is effective on inhibiting the generation of $ONOO^-$, NO and ${\cdot}O_2{^-}$, and that therefore it might have a potential role as a treatment for the inflammatory process and inflammation-related diseases.