• Molecules and Cells
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• 제45권9호
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• pp.649-659
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• 2022

A long-term energy nutritional imbalance fundamentally causes the development of obesity and associated fat accumulation. Lysosomes, as nutrient-sensing and lipophagy centers, critically control cellular lipid catabolism in response to nutrient deprivation. However, whether lysosome activity is directly involved in nutrient-induced fat accumulation remains unclear. In this study, worm fat accumulation was induced by 1 mM glucose or 0.02 mM palmitic acid supplementation. Along with the elevation of fat accumulation, lysosomal number and acidification were also increased, suggesting that lysosome activity might be correlated with nutrient-induced fat deposition in Caenorhabditis elegans. Furthermore, treatments with the lysosomal inhibitors chloroquine and leupeptin significantly reduced basal and nutrient-induced fat accumulation in C. elegans. The knockdown of hlh-30, which is a critical gene in lysosomal biogenesis, also resulted in worm fat loss. Finally, the mutation of aak-2, daf-15, and rsks-1 showed that mTORC1 (mechanistic target of rapamycin complex-1) signaling mediated the effects of lysosomes on basal and nutrient-induced fat accumulation in C. elegans. Overall, this study reveals the previously undescribed role of lysosomes in overnutrition sensing, suggesting a new strategy for controlling body fat accumulation.

• 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.

• 대한한방내과학회지
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• 제32권1호
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• pp.56-67
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• 2011

Objectives : Recent etiological studies show that oxidative stress might play a major role in the diabetes and its complications. Mori Ramulus (MR) has been known to have antioxidative, anti-inflammatory and antidiabetic effects. The methanol extract of MR was tested for its effectiveness in LLC-PK1 cells exposed to high glucose. Methods : The cytoprotective effect of MR was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The antioxidative effect was measured in terms of generation amount of ${\cdot}O_2^-$ by 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), NO by 4,5-diaminofluorescein (DAF-2), $ONOO^-$ by dihydrorhodamine 123 (DHR 123) in the high glucose -treated LLC-$PK_1$ cells. Western blotting was performed using anti-AGE, anti-RAGE, anti-MAPKs(ERK1/2, JNK, p38), anti-PI3K, anti-Akt, and anti-NF-${\kappa}$B (p50, p65) respectively. Results : MR extract reduced cell death and inhibited the generation of ${\cdot}O_2^-$, NO, $ONOO^-$ in the high glucose-treated LLC-$PK_1$ cells. MR inhibited the expression of AGE, RAGE, MAPKs, PI3K, and Akt by means of decreasing NF-${\kappa}$B activation. MR also inhibited NF-${\kappa}$B activation itself. Conclusions : These results indicate MR has cytoprotective, antioxidative, and anti-inflammatory effects. Therefore it is suggested that MR might prevent and cure diabetes and its complications.

• 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.

• 대한환경공학회지
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• 제36권3호
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• pp.172-177
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• 2014

가압형 부상법에 비해 낮은 압력에서 미세기포를 발생시키는 저압형 부상법을 슬러지 부상 농축에 적용하여 그 성능을 검증하고 실제 적용가능성을 확인하였다. 파일롯 규모의 저압형 부상조를 충남 N.S. 하수처리장에 설치하여 혼합슬러지의 농도, 응집제 주입량 및 혼합슬러지 대비 미세기포수의 비율과 같은 운전 변수가 혼합슬러지 농축에 미치는 영향을 확인하였다. 미세기포는 내부 압력이$1.5kgf/cm^2$으로 유지된 미세기포발생기에서 공기와 기포조제가 포함된 물을 고속 충돌 방식으로 발생시켰으며, 이를 부상농축 실험에 사용하였다. 장기운전 시 유입된 혼합슬러지의 SS 농도는 평균 14,400 mg/L였으며, 응집제 농도 27.6 mg/L, 기포조제 농도 4.0 mg/L, 혼합슬러지 대비 미세기포수의 비를 9.7%로 하여 저압형 부유부상조를 운전한 결과, 60,300 mg/L의 농축슬러지 고형물함량과 99% 이상의 고형물 회수율을 얻었다. 이 경우의 고형물 표면적 부하율은 $30kg/m^2/hr$로 2011년 환경부에서 제정한 하수도 상압 부상농축 시설기준 25 $kg/m^2/hr$을 상회하였다.