• 한방재활의학과학회지
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• 제26권3호
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• pp.51-58
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• 2016

Objectives This study was designed to investigate whether the Cheongajihwang-Tang (CT) has an inhibitory effect association with oxidation or inflammation in RAW264.7 cells. Methods Cytotoxic activity of CT extract on RAW264.7 cells was evaluated by using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) solution. Nitric oxide production was measured using Griess reagent system. The total phenolic contents and 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was measured to evaluate the anti-oxidative effects of CT. Dichlorofluorescin diacetate (DCFH-DA) has been used as a substrate for measuring intracellular oxidant production. Results Cheongajihwang-Tang does not impair the cell viability in tested concentration. CT showed anti-oxidative effects in vitro by decreasing electron donating ability, and also showed anti-inflammatory effects suppressing NO and ROS expression in LPS induced RAW264.7 activation. CT inhibited the generation of intracellular ROS production as dose dependant manner. Conclusions CT has anti-oxidative effects and anti-inflammatory activities. These results indicate that CT extract has an anti-inflammatory activities via anti-oxidative effects.

• 대한본초학회지
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• 제23권3호
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• pp.163-173
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• 2008

Objectives : Imyosan (IMS), a drug preparation comprised of Phellodendri Cortex (PC) and Atractylodis Rhizoma (AR), is commonly used as a traditional herbal medicine in Korea and China for the treatment of various inflammatory diseases. However, little is known about the effect of IMS and its component herbs on inflammatory mediators in RAW 264.7 cells. Therefore, in this study, methanol extracts of IMS and its component herbs were examined to determine if they inhibited inflammatory effects in RAW 264.7 cells. Methods : Cytotoxic activity of IJHT and its components on RAW 264.7 cells was using 5-(3-carboxymethoxyphenyl)-2H-tetrazolium 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 : Methanol extract of IMS and its component herbs were significantly reduced iNOS and COX-2 expression as well as NO, PGE2, $IL-1{\beta}$ and IL-6 production in RAW 264.7 cells. Conclusions : The results of this study indicated that the anti-inflammatory effect of Imyosan extract is more potent than that of extracts of its component herbs in macrophages.

• 한방재활의학과학회지
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• 제28권2호
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• pp.37-45
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• 2018

Objectives This study was designed to investigate whether the Gagamtongsoon-San (GT) has an inhibitory effect and its mechanisms are associated with the iNOS and COX-2. Methods Cytotoxic activity of GT extract on RAW264.7 cells was evaluated by using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) solution. Inflammatory condition was induced by LPS. NO production was measured using Griess reagent system. The expressions of iNOS and COX-2 mRNA and protein were determined by realtime PCR. The concentrations of PGE2 were measured by an enzyme immunoassay (EIA). Results The GT does not impair the cell viability in tested concentration $500{\mu}g/ml$ or below. GT significantly reduced the NO production in a dose-dependent manner. GT $500{\mu}g/ml$ also suppressed LPS-induced mRNA expressions of iNOS and COX-2. GT $500{\mu}g/ml$ reduced the PGE2 secretion in LPS induced RAW264.7 cells. Conclusions These outcomes show that GT extract has an anti-inflammatory activities. And also this conclusion can be the data that supports the GT's anti-inflammatory effect objectively.

• 한방안이비인후피부과학회지
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• 제15권2호
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• pp.1-19
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• 2002

Fifteen herbs used in oral medicine were extracted in ethanol and screened for tooth whitening effect, radical scavenging activity, inhibition of inducible nitric oxide synthase activity and anti-bacterial activity on Streptoccocus mutans. The results are as followed: 1. The tooth whitening effect of toothpaste containing 1 $\%$ ethanol extracts of 15 herbs was tested in brushing method using artificially stained HAT tablet and bovine tooth specimens. Toothpaste containing 1$\%$ of Vucia unijuga, Illicium verum, Angelica dahurica and Piper longum extracts showed tooth whitening effects on both HAT tablet and bovine tooth specimens. But the others did not show a considerable activity. 2. The antioxidant activity of ethanol extracts of 15 herbs was tested using the method of 1,1-diphenyl-2-picryl hydrazyl (DPPH) reactivity. Two ethanol extracts of Cimicifuga heracleifolia and Phyllostachys nigra var. henonis were found to be the most effective on radical scavenging activity. C. heracleifolia and P. nigra var. henonis extracts removed 86$\%$ and 81 $\%$ of DPPH radical at 0.01$\%$, compared with butylated hydroxy toluene as positive control. 3. The inhibition activity of ethanol extracts on nitrate production In RAW264.7 cell stimulated by lipopolysacchride was tested using the Griess reagent. But all extracts did not inhibit nitrate production. 4. The antibacterial activity on Streptoccocus mutans was tested by paper disk method. But no one extract showed any anti - bacterial activity.

• 대한의생명과학회지
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• 제24권4호
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• pp.365-371
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• 2018

In this study, antioxidant and anti-neuroinflammatory of ethanol extracts from walnuts's (Juglans regia L.) shell were investigated in vitro. Radical-scavenging activities of the walnuts's shell ethanol extracts (WSE) were examined by using ABTS radicals and ${\alpha},{\alpha}$-diphenyl-${\beta}$-picrylhydrazyl (DPPH) radicals assay. In the ABTS and DPPH radical scavenging activity, $RC_{50}$ of WSE were measured as 15.74 and $40.13{\mu}g/mL$, respectively. Also, to evaluate the anti-neuroinflammatory effects of WSE in lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. The production of proinflammatory cytokines NO were examined by LPS in BV-2 cell. BV-2 cells activated with LPS were treated with various doses (10, 25, 50, $100{\mu}g/mL$) of WSE. Supernatants were analyzed for the production of NO using Griess reagent. WSE up to $10{\mu}g/mL$ still required to inhibit NO induced by LPS. These results showed that walnuts's (Juglans regia L.) shell can be used as an easily accessible source of natural anti-neuroinflammatory and natural antioxidants.

• Journal of Animal Science and Technology
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• 제63권5호
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• pp.1159-1168
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• 2021

Ovotransferrin (OTF), an egg protein known as transferrin family protein, possess strong antimicrobial and antioxidant activity. This is because OTF has two iron binding sites, so it has a strong metal chelating ability. The present study aimed to evaluate the improved immune-enhancing activities of OTF hydrolysates produced using bromelain, pancreatin, and papain. The effects of OTF hydrolysates on the production and secretion of pro-inflammatory mediators in RAW 264.7 macrophages were confirmed. The production of nitric oxide (NO) was evaluated using Griess reagent and the expression of inducible nitric oxide synthase (iNOS) were evaluated using quantitative real-time polymerase chain reaction (PCR). And the production of pro-inflammatory cytokines (tumor necrosis factor [TNF]-α and interleukin [IL]-6) and the phagocytic activity of macrophages were evaluated using an ELISA assay and neutral red uptake assay, respectively. All OTF hydrolysates enhanced NO production by increasing iNOS mRNA expression. Treating RAW 264.7 macrophages with OTF hydrolysates increased the production of pro-inflammatory cytokines and the phagocytic activity. The production of NO and pro-inflammatory cytokines induced by OTF hydrolysates was inhibited by the addition of specific mitogen-activated protein kinase (MAPK) inhibitors. In conclusion, results indicated that all OTF hydrolysates activated RAW 264.7 macrophages by activating MAPK signaling pathway.

• 한방안이비인후피부과학회지
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• 제35권1호
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• pp.11-23
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• 2022

Objectives : The purpose of this study is to confirm the anti-inflammatory effects of Cistanche deserticola Water Extracts(CDWE). Methods : In this study, cell viability was determined by MTT assay. NO production was determined with Griess reagent, and IL-6 production was determined by ELISA. And western blot analysis was performed to confirm the protein expression of NOS2, COX-2, I𝜅B-𝛼, p-I𝜅B-𝛼, and NF-𝜅B. Results : CDWE was not cytotoxic at 15.625-1,000㎍/㎖ in RAW 264.7 cells. CDWE inhibited NO production in a concentration-dependent manner, with inhibitory effect on expression levels of IL-6. Expression level of NOS2, NF-𝜅B p65, and p-I𝜅B-𝛼 decreased at 250-1,000㎍/㎖. mRNA expression of COX-2 suppressed at 250, 1,000㎍/㎖. Conclusions : These results suggest that CDWE has anti-inflammatory effects and can be used as an anti-inflammatory therapeutic material.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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• pp.176-176
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• 1998

An enhanced formation of nitric oxide (NO) is an important mediator of hypotention, peripheral vasodilation and vascular hyporeactivity to vasoconstrictor agents in endotoxaemia. And tumor necrosis factor (TNF${\alpha}$), as a primary mediator of circulatory shock has been known to induce inducible nitric oxide synthase (i-NOS), leading to excessive production of NO. We isolated two sesquiterpene lactone compounds from Saussurea lappa and their structures were elucidated as dehydrocostus lactone and costunolide. These compounds inhibit the production of both NO and TNF${\alpha}$ by LPS (1 $\mu\textrm{g}$/$m\ell$)-activated Raw 264.7 cells. NO was measured spectropho-tometrically as nitrite by the Griess reagent and TNF${\alpha}$ by ELISA. Dehydrocostus lactone (IC$\sub$50/ : 3.0 ${\mu}$M) and costunolide (IC$\sub$50/ : 4.5 ${\mu}$M) inhibited the production of NO in LPS-activated Raw 264.7 cells by suppressing inducible nitric oxide synthase enzyme expression. These compounds also decreased the TNF${\alpha}$ levels in LPS-activated system in vitro and in vivo.

• 대한본초학회지
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• 제24권3호
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• pp.97-102
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• 2009

Objectives : We determined the anti-inflammatory activity of KCNS-001 that is a herbal formula including 6 medicinal plants and that are used to mitigate atopic dermatitis in oriental medicine. Methods : To evaluate anti-inflammatory effect of KCNS-001, we measured the production of reactive oxygen species (ROS), nitric oxide (NO) and cyclooxygenase-2 (COX-2) in LPS-activated Raw 264.7 cells. Cell viability was determined by MTT assay. The concentrations of ROS and relative level of NO were measured with DPPH assay and Griess reagent, respectively. COX-2 and TNF-$\alpha$ were detected by enzyme immuno assay (EIA) and enzyme-linked immunosorbent assay (ELISA). Results : ROS and NO production were reduced by KCNS-001 in a dose-dependent manner. KCNS-001 significantly inhibited activity of COX-2 and suppressed the release of tumor necrosis factor-alpha (TNF-$\alpha$). Conclusions : These results indicate that the KCNS-001 may have an anti-inflammatory agent for the treatment of various inflammatory disease.

• 한국식생활문화학회지
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• 제38권5호
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• pp.365-372
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• 2023

Ipomoea aquatic is a leafy vegetable of the Convolvulaceae family, and is a tropical plant widely inhabiting southern China and Southeast Asia, and is widely known as Morning Glory in the West. In this study, the anti-inflammatory effects of ethyl acetate extract from Ipomoea aquatic extracts (IAE) were tested against lipopolysaccharide (LPS)-induced activation microglia BV2 cells. The production of nitric oxide (NO) and cell viability were measured using the Griess reagent and MTT assay, respectively. Inflammatory cytokine [interleukin (IL)-6, tumor necrosis factor (TNF)-α, and interleukin-1β (IL-1β)] were detected qPCR in LPS induced BV-2 cells. Subsequently, nuclear factor (NF)-κB, mitogen-activated protein kinases (MAPKs), and nuclear factor erythroid-2-related factor 2 (Nrf2) were analyzed through western blot analyses and immunofluorescence. Ipomoea aquatic down-regulated of inflammatory markers and up-regulated anti-inflammatory and anti-oxidants in BV2 cells.