• Biomolecules & Therapeutics
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• 제17권3호
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• pp.293-298
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• 2009

Korean mistletoe lectin (KML) is the major component found in Viscum album var. (coloratum), displaying anti-cancer and immunostimulating activities. Even though it has been shown to boost host immune defense mechanisms, the regulatory roles of KML on the functional activation of macrophages have not been fully elucidated. In this study, regulatory mechanism of KML on macrophage-mediated immune responses was examined in terms of KML-mediated signaling event. KML clearly induced mRNA expression of tumor necrosis factor (TNF)-$\alpha$, the generation of reactive oxygen species (ROS) and phagocytic uptake in RAW264.7 cells. All of these events were strongly suppressed by U0126, whereas TNF-$\alpha$ mRNA was not diminished by SB203580 and SP600125, indicating ERK as a central enzyme managing KML-induced up-regulation of macrophage functions. Indeed, KML strongly induced the phosphorylation of ERK in a time-dependent manner without altering its total level. Therefore, these data suggest that ERK may be a major signaling enzyme with regulatory property toward various KML-mediated macrophage responses.

• 대한본초학회지
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• 제25권4호
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• pp.103-112
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• 2010

Objective : The aim of this study was to determine whether methanol extract of Do-Ki-Tang (DKT) inhibit free radical generation and production of nitrite an index of NO, $PGE_2$, iNOS, COX-2 and pro-inflammatory cytokines such as TNF-${\alpha}$, IL-$1{\beta}$, IL-6 and MCP-1 in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Methods : Cytotoxic activity of extract on RAW 264.7 cells was measured using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. The expression level of inflammatory response-related proteins was confirmed by western blot. The production of proinflammatory cytokines was measured by ELISA. Results : Our results indicated that DKT scavenged DPPH radical and nitric oxide in vitro. Moreover, DKT significantly inhibited the LPS-induced NO, $PGE_2$ production and iNOS, COX-2 expression accompanied by an attenuation of TNF-${\alpha}$, IL-$1{\beta}$, IL-6 and MCP-1 formation in macrophages. Furthermore, DKT treatment also blocked LPS-induced intracellular ROS production and the activation of NF-${\kappa}B$ and MAPKs. Conclusion : Our data suggest that the anti-inflammatory effect of DKT is mediated through down-modulation of pro-inflammatory mediators and cytokines by blocking the signaling pathways of NF-${\kappa}B$ and MAPKs. These inhibitory effects by DKT represent a potential therapeutic approach to the treatment of inflammatory diseases.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 추계학술대회
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• pp.111-111
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• 2018

Aronia (black chokeberry), a species of berries is source to a very large number of bioactive compounds like polyphenols, flavonoids, anthocyanins, and tannins in comparison to any other species. Owing to its antioxidant, anti-carcinogenic, anti-aging and anti-inflammatory properties. Fermentation- a bioconversion process exploiting the biological metabolic reaction of micro-organisms, has several benefits like improving the efficacy and safety of physiologically active substances, generation of new functional material, improving the adsorption rate and many others. Antigens like pathogens, food, pollen etc., generate a protective immune response in body tissues, and the process be referred to as inflammation. However, this when excessive results in a condition referred to as refractory inflammatory disease, whose incidence is increasing in the recent times, especially amongst children. The current study intended to assess the anti-oxidant activity, presence of polyphenols and anti-inflammatory efficacy of Aronia extract fermented by Lactic acid bacteria isolated from fermented sea foods. Aronia fruits collected from Sunchang, Chonbuk were lyophilized for fermentation. So as to maximise the efficacy of the fermented Aronia extract, the quantitative effects of lactic acid bacteria species, composition of extraction solution, influence of temperature and time on antioxidant activity and total polyphenol contents were investigated using an experimental design. Anti-inflammatory activity was evaluated on NO and cytokine ($TNF-{\alpha}$, IL-6) production in LPS activated Raw 264.7 cells. Results indicated that antioxidant effect and total polyphenol contents were best improved in extract of Aronia fermented by P. pentosaceus. In addition, NO and cytokine ($TNF-{\alpha}$, IL-6) levels were decreased significantly after fermentation. Thus, it was found that the anti-inflammatory activity of Aronia greatly increased after fermentation process using P. pentosaceus.

• 대한수의학회지
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• 제55권2호
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• pp.81-88
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• 2015

This study was conducted to isolate lactic acid bacteria (LAB) from dog intestine and identify potential probiotic strains for canine use. One hundred and one LAB were isolated from feces of 20 healthy dogs. Acid, bile, and heat resistance along with adherence to Caco-2 cells and antimicrobial activity against pathogens were examined. To analyze immunomodulative effects, the production of nitric oxide (NO), TNF-${\alpha}$, and IL-$1{\beta}$ was measured using RAW 264.7 macrophages. Additionally, RAW BLUE cells were used to evaluate nuclear factor-${\kappa}B$ (NF-${\kappa}B$) generation. Ultimately, three strains were selected as canine probiotics and identified as Lactobacillus reuteri L10, Enterococcus faecium S33, and Bifidobacterium longum B3 by 16S rRNA sequence analysis. The L10 and S33 strains showed tolerance to pH 2.5 for 2 h, 1.0% Oxgall for 2 h, and $60^{\circ}C$ for 5 min. These strains also had strong antimicrobial activity against Escherichia coli KCTC 1682, Salmonella Enteritidis KCCM 12021, Staphylococcus aureus KCTC 1621, and Listeria monocytogenes KCTC 3569. All three strains exerted better immunomodulatory effects than Lactobacillus rhamnosus GG (LGG), a well-known commercial immunomodulatory strain, based on NO, NF-${\kappa}B$, IL-$1{\beta}$, and TNF-${\alpha}$ production. These results suggested that the three selected strains could serve as canine probiotics.

• 대한방사선기술학회지:방사선기술과학
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• 제39권3호
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• pp.443-449
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• 2016

본 연구에서는 Candida albicans로 유도한 염증반응에 대해 스쿠알렌의 처치를 통해 염증반응을 경감시키는가에 대해 확인하였다. 실험동물은 (ICR계 생쥐) 각 실험군별 7마리씩을 사용하여 생체 내 실험을 하였다. Candida albicans-유도 염증인자 중 종양괴사인자-알파, 인터루킨-6, 산화질소는 ELISA kits 방법을 이용하여 관찰하였다. 실험을 통해 다음과 같은 결론을 얻었다. 1. Candida albicans를 감염시킨 군에서 1, 3일째 모두 신장조직 내에서의 산화질소 생성량을 감소시키는 것을 확인할 수 있었다 (p< 0.05). 2. 스쿠알렌 (80ml/kg)을 7일간 1일 1회 전 처치(복강투여)한 다음, Candida albicans를 감염시킨 군에서 3일째 군에서만 신장 조직 내에서의 종양괴사인자-알파 생성량을 감소시키는 것을 확인할 수 있었다(p< 0.05). 3. 스쿠알렌 (80m/kg)을 7일간 1일 1회 전 처치(복강투여)한 다음. Candida albicans를 감염 시킨 군에서 3일째 군에서만 신장 조직 내에서의 인터루킨-6 생성량을 감소시키는 것을 확인할 수 있었다(p< 0.05). 결론적으로 스쿠알렌이 Candida albicans-유도 염증인자들의 억제를 위해서는 예방적 차원에서 스쿠알렌을 공급하는 것이 효과가 있음을 알 수 있었다. 따라서 스쿠알렌 처치가 Candida albicans-유도 면역억제에 대해 회복 효과를 기대할 수 있을 것으로 사료된다.

• 대한본초학회지
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• 제25권2호
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• pp.129-136
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• 2010

Objective : Sinapis alba L. (SA) is a korean traditional herbal medicine that is usually used to prevent or treat inflammatory diseases, such as respiratory infection and rheumatoid arthritis. However, the effects of SA supplementation in vitro on serum antibody levels, splenocyte and peritoneal macrophage immune responses have not yet been determined. In this study, we examined the effect of SA on the production of Th1/Th2 cytokines. Methods : Splenocytes were isolated from naive C57BL/6 mice. Cells were enriched for CD4+ cell populations by first staining the cells with anti-CD4 (BD PharMingen, Calif, USA). CD4+ T cells were selected on a (CS) column, and the flow-through was collected as CD4+ T cells. Isolated cells were activated by overnight incubation on 24-well plates coated with $1{\mu}g/mL$ anti-CD3, $1{\mu}g/mL$ anti-CD28 and with SA ($100{\mu}g/mL$). Primary macrophages were collected from the peritoneal cavities of mice (8-week-old female C57BL/6). The peritoneal macrophages were washed and plated with RPMI-1640 overnight for the experiments. After 48-hours cultures, samples were centrifuged at 2000 rpm for 10 minutes, and the supernatants were stored at $-80^{\circ}C$. Mouse IL-4, IFN-$\gamma$ and TNF-$\alpha$ were quantified using ELISA kits (BioSource International, Camarillo, Calif, USA) according to the manufacturer's protocols. Results : SA at 100ug/ml decreased the generation of Th1 cytokine (IFN-$\gamma$) by 0.5-fold. However, SA has no effect on Th2 (IL-4) production. Conclusions : These results suggest that SA may play an important role in the control of T-cell-mediated autoimmunity by down-regulation of Th1 cytokine (especially IFN-$\gamma$, TNF-$\alpha$). These data may contribute to the design of new immunomodulating treatments for a group of autoimmune diseases.

• 한국식품영양과학회지
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• 제41권10호
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• pp.1371-1377
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• 2012

산양삼 55% 에탄올 추출물의 항산화 효과 및 LPS로 염증이 유도된 RAW 264.7 대식세포에서 염증매개 물질의 억제능력을 측정하여 항염증 효과를 살펴보았다. 폴리페놀 함량, DPPH, ABTS sytem 및 SOD 유사활성을 측정하여 항산화 효과를 확인한 결과, 높은 폴리페놀 함량(357.45 mg/100 g)이 확인되었고, free radical 소거활성을 측정한 결과 KPGE는 농도 의존적으로 free radical을 소거하였으며, 대조군인 BHT와 유사한 소거활성이 확인되어 KPGE의 우수한 항산화 활성을 확인할 수 있었다. 조사된 최고 농도(1,000 ${\mu}g/mL$)에서 DPPH(80%)와 ABTS(86%)는 가장 높은 활성을 나타냈다. SOD 유사활성은 1, 10, 100 ${\mu}g/mL$ 농도에서 22%의 활성이 확인되었고, 500, 1000 ${\mu}g/mL$ 농도에서 33%의 높은 항산화 활성이 확인되었다. 또한, KPGE의 항염증 효과를 확인하기 위해 LPS로 염증이 유도된 RAW 264.7 대식세포에서 NO, $PGE_2$ 및 전염증성 cytokine(TNF-${\alpha}$, IL-$1{\beta}$, IL-6)을 측정한 결과, 추출물은 농도 의존적으로 NO, $PGE_2$ 생성 및 전염증성 cytokine인 TNF-${\alpha}$, IL-$1{\beta}$, IL-6의 생성을 효과적으로 억제하는 것을 확인할 수 있었다. 본 연구 결과를 통하여 산양삼 55% 에탄올 추출물 KPGE는 높은 항산화 활성과 염증매개 물질의 생성을 억제하는 우수한 항염증 효과가 있는 것으로 확인되었다.

• BMB Reports
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• 제40권5호
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• pp.636-643
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• 2007

This study was designed to identify and characterize the mechanism of macrophage activation by AAP, an acidic polysaccharide fraction isolated from the roots of Angelica sinensis (Oliv.) Diels. As a result, AAP significantly enhanced nitric oxide (NO) production and cellular lysosomal enzyme activity in murine peritoneal macrophages in vitro and in vivo. Furthermore, L-NAME, a specific inhibitor of inducible nitric oxide synthase (iNOS), effectively suppressed AAP-induced NO generation in macrophages, indicating that AAP stimulated macrophages to produce NO through the induction of iNOS gene expression and the result was further confirmed by the experiment of the increase of AAP-induced iNOS transcription in a dose-dependent manner. To further investigate, AAP was shown to strongly augment toll-like receptor 4 (TLR4) mRNA expression and the pretreatment of macrophages with anti-TLR4 antibody significantly blocked AAP-induced NO release and the increase of iNOS activity, and tumor necrosis factor-$\alpha$ (TNF-$\alpha$) secretion.

• Preventive Nutrition and Food Science
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• 제16권4호
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• pp.299-306
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• 2011

The present study was designed to evaluate the inhibitory effects of fermented Liriope platyphylla extract on the production of inflammation-related mediators (NO, ROS, NF-${\kappa}B$, iNOS and COX-2) and pro-inflammatory cytokines (TNF-${\alpha}$, IL-$1{\beta}$, IL-6) in lipopolysaccharide-stimulated RAW 264.7 macrophages. Freeze-dried Liriope platyphylla was fermented by Saccharomyces cerevisiae and extracted with 70% ethanol. In lipopolysaccharide-stimulated macrophage cells, the treatment with fermented Liriope platyphylla extract decreased the generation of intracellular reactive oxygen species dose-dependently and increased antioxidant enzyme activities, including superoxide dismutase, catalase and glutathione peroxidase. Fermented Liriope platyphylla extract also inhibited NO production in lipopolysaccharide-stimulated RAW 264.7 cell. The expressions of NF-${\kappa}B$, iNOS, COX-2 and pro-inflammatory cytokines were inhibited by the treatment with fermented Liriope platyphylla extract. Thus, this study shows the fermented Liriope platyphylla extract could be effective at inhibiting the inflammation process.

• 대한본초학회지
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• 제23권4호
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• pp.179-189
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• 2008

Objectives: Effects of Rhodiola rosea root on immunity were examined. Methods: Water extracts and methanol extracts of Rhodiola rosea root were treated on the increase of immunity cells and the activation of cytokines were examined. Results: 1. Water extracts and methanol extracts of Rhodiola rosea root had positive effects on the increase of B-cells, T-cells, and NK cells, and on the facilitation of cytokines such as IL-6 and TNF-${\alpha}$. 2. Water extracts and methanol extracts of Rhodiola rosea root activated NK cells, and induced the generation of NO- of NK cells and macrophages. Conclusions: Rhodiola rosea root can be used to increase immunity.