• 대한한의학회지
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• 제28권2호통권70호
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• pp.155-165
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• 2007

Objective : The Angelica gigas Nakai ethanol extract (AGE) was investigated to compare nitric oxide (NO) production and $NF-{\kappa}B$ activity from RAW 264.7 cells, since NO and nuclear $factor-{\kappa}B$ $(NF-{\kappa}B)$ have been shown to be factors implicated in inflammatory disease. Method : AGE was prepared by extracting medicinal herb with 70% (v/v) ethanol solution. We investigated production of nitric oxide (NO) and inducible nitric oxide synthase (iNOS) gene expression by ARE in LPS-stimulated RAW 264.7 macrophage cells. We also investigated inhibition of LPS-induced activation of $NF-{\kappa}B$ on western blot. Result : LPS-induced RAW 264.7 cells increased NO production and iNOS expression. Upon treatment with AGE, nitrite production was significantly inhibited in a concentration-dependent manner compared to the untreated control. AGE inhibited this LPS-induced iNOS mRNA and protein in a dose-dependent manner. AGE markedly inhibited the expression of iNOS mRNA and protein at a concentration of 100 ${\mu}g/ml$. LPS-induced RAW 264.7 cells with AGE blocked inhibitory $factor-{\kappa}B{\alpha}$ degradation. Conclusion :This study shows that AGE seems to attenuate inflammation through inhibition of NO production and iNOS expression by blockade of $NF-{\kappa}B$ activation in LPS-stimulated RAW 264.7 cells.

• Tuberculosis and Respiratory Diseases
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• 제47권2호
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• pp.172-183
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• 1999

연구배경: 산화질소(${\cdot}NO$)는 여러 세포에서 산화질소 합성효소(NOS)에 의해서 생산되며 다양한 병태생리과정에 관여한다. 여러 cytokine들이 iNOS의 발현을 촉진시키고 산화질소 생산을 증가시킴으로써 염증반응을 증폭시키고 세포와 조직손상을 초래한다고 알려진 바, 과산화수소($H_2O_2$)가 세포내 NOS의 발현과 산화질소형성에 미치는 영향을 알아보고자 하였다. 방법: 마우스 대식세포주 RAW264.7에 여러 가지 cytokine과 세균 내독소 (LPS)로 자극을 준 세포군 이에 더하여 $H_2O_2$, NOS 억제제 (L-NAME) 및 항산화제 (catalase)등을 사용하여 세포를 자극한 후 생성된 산화질소 산화물의 농도를 측정하고 Northern analysis로 iNOS mRNA의 발현정도를 보아 다음과 같은 성적을 얻었다. 결과: Cytokine과 LPS 자극군에서 대조군보다 ${\cdot}NO$ 생산이 높았고, 이 자극군에 $H_2O_2$를 추가로 자극하였을 때 ${\cdot}NO$생산이 2 배 이상 유의하게 높았다. Cytokine 자극군에서 $H_2O_2$의 자극 농도에 따른 ${\cdot}NO$생산은 $H_2O_2$의 농도가 증가할수록 유의하게 증가하였다. LPS와 IFN-$\gamma$ 자극군에서 L-NAME을 같이 자극시에 ${\cdot}NO$의 양은 L-NAME의 농도증가에 따라 유의하게 감소하였고, Cytokine 및 $H_2O_2$자극군에서도 추가로 자극한 L-NAME 의 농도증가에 따라 ${\cdot}NO$의 양은 유의하게 감소하였다. Cytokine과 $H_2O_2$ 자극균에 catalase를 같이 자극 하였을 때 ${\cdot}NO$의 양은 유의하게 감소했고, Mercaptoethanol과 phenanthroline을 전처치하고 LPS와 IFN-$\gamma$ 및 $H_2O_2$로 자극한 군에서 이들의 전처치한 농도가 높을수록 ${\cdot}NO$의 양은 유의하게 Cytokine자극군과 IFN-$\gamma$, LPS 자극군에 $H_2O_2$를 추가 자극 후 Northern analysis 결과 $H_2O_2$는 iNOS mRNA 발현을 현저히 증가시켰다. 결론: 이상의 결과로 과산화수소가 cytokine과 내독소 등으로 자극된 마우스 대식세포에서 산화질소생산에 유의한 증폭효과를 나타냈고, iNOS mRNA 의 발현도 증가시켰음을 확인할 수 있었다.

• 대한본초학회지
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• 제23권2호
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• pp.169-178
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• 2008

Objectives : Inflammation is important event in the development of vascular diseases including hypertension, atherosclerosis, and restenosis. Injinho-tang(IJHT) has been used as a traditional Korean herbal medicine since ancient times, and today it is widely used as a medication for jaundice associated with inflammation of the liver. The aim of this study was to determine whether IJHT and its components inhibit production of nitrite, an index of NO, and proinflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. Methods : Cytotoxic activity of IJHT and its components on RAW 264.7 cells was using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. The nitric oxide (NO) production was measured by Griess reagent system. And proinflammatory cytokines were measured by ELISA kit. The levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were detected by western blot. Results : IJHT and its components significantly inhibited the LPS-induced NO production and iNOS expression accompanied by an attenuation of tumor necrosis factor-alpha (TNF-${\alpha}$), interleukin-6 (1L-6), IL-$1{\beta}$ and monocyte chemoattractant protein-1 (MCP-1) formation in macrophages. Conclusions : IJHT and its components inhibit LPS-induced inflammation via decreasing cytokines production. These results indicate that IJHT and its components have potential as an anti-inflammation and anti-artherosclerosis agent.

• 대한한의학방제학회지
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• 제18권1호
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• pp.169-179
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• 2010

The pistil of nelumbo nucifera (PNN) is used in the treatment of nocturnal pollution, hematemesis, epistaxis, metrorrhagia and diarrhoea in traditional medicine. The present study was examined to evaluate the effects of PNN on the production of pro-inflammatory mediators in vitro. After the treatment of PNN, cell viability was measured by MTT assay, nitric oxide (NO) production was monitored by measuring the nitrite content in culture medium. The protein bands were determined by immunoblot analysis and levels of cytokines were analyzed by sandwich immunoassays. In the MTT assay, the doses of PNN extract (0.03, 0.10 mg/ml) had no significant cytotoxicity. The increases of NO production and inducible nitric oxide synthase expression were detected in lipopolysaccharide(LPS)-activated Raw 264.7 cells compared with control, in contrast, these increases were significantly attenuated by pre-treatment with PNN. In cytokine assay, the massive pro-inflammatory cytokines such as tumour necrosis factor-$\alpha$, interleukin (IL)-$1{\beta}$ and IL-6 were induced in LPS-activated Raw 264.7 cells, but pre-treatment of Raw 264.7 cells with PNN caused inhibition (TNF-$\alpha$=14.17%, IL-$1{\beta}$=107.43%, IL-6=46.27%) the production of cytokines by LPS. In addition, PNN reduced prostaglandin E2 productions in a dose-dependent manner (0.03mg/ml=37.52%, 0.10 mg/ml=83.77%) as a consequence of the inhibition of cyclooxygenase-2 expression. Taken together, our data indicates that PNN can regulate the inflammatory response in macrophage cells activated by Gram-negative infection.

• The Korean Journal of Physiology and Pharmacology
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• 제4권4호
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• pp.325-331
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• 2000

Tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ plays important roles in inflammatory responses. Some of tetrahydroisoquinoline (THI) compounds exhibited to inhibit iNOS expression in animal studies and RAW 264.7 cells, but the action of THI on inflammatory reaction was not fully investigated. In the present study, we examined a limited series of THIs (higenamine, YS-51 and THI-52) on the $TNF-{\alpha}$ mRNA expression in mouse peritoneal macrophages by Northern analysis. When thioglycollate-stimulated peritoneal macrophages were incubated with LPS (100 ng/ml), expression of $TNF-{\alpha}$ mRNA was evident and reached its maximum at 2.5 h, which was reduced concentration-dependently by treatment with THIs. When the $TNF-{\alpha}$ activity of macrophage-conditioned media was measured using a TNF-sensitive L929 fibroblast cell line, CCL 1, all THIs increased the cell viability in a concentration dependent manner. The concentrations of THIs used are not cytotoxic by itself when analysed by MTT. Furthermore, nitrite/nitrate level was significantly reduced by the presence of THIs in cells treated with $LPS+interferon-{\gamma}\;(IFN-{\gamma}).$ It is concluded, thus, that these results strongly indicated that THIs can suppress the $TNF-{\alpha}$ expression and reduce NO, which may be useful for the inflammatory disorders.

• The Korean Journal of Physiology and Pharmacology
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• 제18권6호
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• pp.481-487
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• 2014

Ischemic postconditioning (IPost) could decrease ischemia-reperfusion (IR) injury. It has not yet reported whether IPost is useful when ischemic heart disease is accompanied with co-morbidities like hyperthyroidism. The aim of this study was to examine the effect of IPost on myocardial IR injury in hyperthyroid male rats. Hyperthyroidism was induced with administration of thyroxine in drinking water (12 mg/L) over a period of 21 days. After thoracotomy, the hearts of control and hyperthyroid rats were perfused in the Langendorff apparatus and subjected to 30 minutes global ischemia, followed by 120 minutes reperfusion; IPost, intermittent early reperfusion, was induced instantly following ischemia. In control rats, IPost significantly improved the left ventricular developed pressure (LVDP) and ${\pm}dp/dt$ during reperfusion (p<0.05); however it had no effect in hyperthyroid rats. In addition, hyperthyroidism significantly increased basal $NO_x$ (nitrate+nitrite) content in serum ($125.5{\pm}5.4{\mu}mol/L$ vs. $102.8{\pm}3.7{\mu}mol/L$; p<0.05) and heart ($34.9{\pm}4.1{\mu}mol/L$ vs. $19.9{\pm}1.94{\mu}mol/L$; p<0.05). In hyperthyroid groups, heart $NO_x$ concentration significantly increased after IR and IPost, whereas in the control groups, heart $NO_x$ were significantly higher after IR and lower after IPost (p<0.05). IPost reduced infarct size (p<0.05) only in control groups. In hyperthyroid group subjected to IPost, aminoguanidine, an inducible nitric oxide (NO) inhibitor, significantly reduced both the infarct size and heart $NO_x$ concentrations. In conclusion, unlike normal rats, IPost cycles following reperfusion does not provide cardioprotection against IR injury in hyperthyroid rats; an effect that may be due to NO overproduction because it is restored by iNOS inhibition.

• 상하수도학회지
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• 제23권5호
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• pp.547-555
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• 2009

The objectives of this research were focused on the effects of various operating parameters on nitrous oxide emission such as C/N ratio, ammonia concentration and HRT in the hybrid and suspension reactors. With the decreasing of C/N ratios, $N_2O$ emission rates in the both processes were increased because organic carbon source for denitrification was depleted. In case of biofilm reactor operated using medium, $N_2O$ release from the nitrification was not affected by the variation of ammonia concentration. But in the suspension reactor, $N_2O$ production from the nitrification was rapidly increased with the increase of ammonia. Nitrite accumulation caused by undesirable nitrification conditions could be a important reason for the increase in the $N_2O$ production from the aerobic reactor. And rapid increase in $N_2O$ production was reflected by the decrease of HRT, similar to the results observed in the results of ammonia loading changes. So it could be said that it is very important to put in consideration both its optimum conditions for wastewater treatment efficiency and suitable conditions for $N_2O$ diminish, simultaneously, in order to development an eco-friendly and advanced wastewater treatment, especially in BNR process.

• Asian Pacific Journal of Cancer Prevention
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• 제15권4호
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• pp.1807-1810
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• 2014

Background: It is known that inducible nitric oxide synthase (iNOS)/nitric oxide (NO) plays an integral role during intestinal inflammation, an important factor for colon cancer development. Natural compounds from Curcuma longa L. (Zingiberaceae) have long been a potential source of bioactive materials with various beneficial biological functions. Among them, a major active curcuminoid, demethoxycurcumin (DMC) has been shown to possess anti-inflammatory properties in lipopolysaccharide (LPS)-activated macrophages or microglia cells. However, the role of DMC on iNOS expression and NO production in an in vitro inflamed human intestinal mucosa model has not yet been elucidated. This study concerned inhibitory effects on iNOS expression and NO production of DMC in inflamed human intestinal Caco-2 cells. An in vitro model was generated and inhibitory effects on NO production of DMC at 65 ${\mu}M$ for 24-96 h were assessed by monitoring nitrite levels. Expression of iNOS mRNA and protein was also investigated. DMC significantly decreased NO secretion by 35-41% in our inflamed cell model. Decrease in NO production by DMC was concomitant with down-regulation of iNOS at mRNA and protein levels compared to proinflammatory cytokine cocktail and LPS-treated controls. Mechanism of action of DMC may be partly due to its potent inhibition of the iNOS pathway. Our findings suggest that DMC may have potential as a therapeutic agent against inflammation-related diseases, especially in the gut.

• 한방안이비인후피부과학회지
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• 제28권4호
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• pp.51-61
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• 2015

Objective : This research has been conducted to comparative analysis of anti-inflammatory effects to suggest a usefulness of ethanol extracts fromGentianae sino-ornata(GSO) as a functional material for inflammatory activities.Methods : Cell viability was measured by neutral red (NR) assay, and nitric oxide (NO) production in RAW264.7 cells was monitored by measuring the nitrite content in culture medium. The expressions of cyclooxigenase-2 (COX-2) was determined by western blot analysis, and Inducible nitric oxide syntase (iNOS) and cytokine were determined by reverse transcriptase-polymerase chain reaction (RT-PCR).Results : When the GSO extract was added the concentration of 5-20 ㎍/㎕, the viability of cells was maintained 90% or more at all levels. NO production was suppressed by the treatment of GSO in LPS-stimulated RAW264.7 cells. GSO inhibited the expression of iNOS, COX-2, IL-1βand IL-6 in LPS-stimulated RAW264.7 cells.Conclusions: From this results, we consider that GSO can be a useful therapeutic and preventive approach to various inflammatory diseases as a functional material for inflammatory activities.

• 융합정보논문지
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• 제10권2호
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• pp.89-95
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• 2020

아로니아 비가식 부위(잎, 송이가지, 가지)를 바이오 기능성 소재로 활용하기 위하여 부위별, 용매별 추출물의 총 폴리페놀 함량과 생리활성을 분석하였다. 잎과 가지 추출물의 폴리페놀 함량은 열매와 송이가지 추출물보다 더 높은 값을 나타냈고, 아로니아의 부위별 항산화능(DPPH 래디컬과 ABTS 래디컬 소거능)을 분석한 결과, 열수추출물과 메탄올추출물 두 경우에서 가지, 잎, 열매, 송이가지의 순으로 활성을 나타냈다. Tyrosinase 저해활성을 평가한 결과, 열수추출물은 열매, 잎, 가지, 송이가지의 순으로 높은 저해활성을 보였으며, 메탄올추출물의 경우 가지, 잎, 열매, 송이가지의 순으로 저해활성을 갖는 것으로 나타났다. Nitric oxide 저해활성은 열수추출물과 메탄올 추출물 두 경우에서 열매, 잎, 가지, 송이가지의 순으로 높았다. 결론적으로 아로니아 가지와 잎은 항산화, 미백 및 항염증 활성을 갖고 있는 것으로 확인하였고, 추가적인 연구를 통하여 화장품 등의 바이오 헬스케어 제품의 기능성 바이오소재로 개발할 수 있을 것으로 기대된다.