• 한국식품영양과학회지
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• 제45권2호
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• pp.194-201
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• 2016

본 연구에서는 lipopolysaccharide(LPS)로 자극한 마우스 대식세포인 RAW 264.7 세포에서 가락진두발 에탄올 추출물(CNYEE)의 항염증 효과를 알아보기 위하여 nitric oxide(NO)와 pro-inflammatory cytokine의 분비량을 확인하였다. 그 결과 CNYEE 모든 농도에서 LPS만을 처리한 대조군과 비교하였을 때 유의성 있게 NO와 pro-inflammatory cytokine의 분비량을 저해하였으며, 특히 $100{\mu}g/mL$ 농도에서는 IL-6의 분비량을 70% 이상 억제하였고, TNF-${\alpha}$ 및 IL-$1{\beta}$의 분비량은 50% 이상의 억제 효과를 나타내었다. CNYEE에 의한 염증매개물질의 분비 감소가 전사인자인 nuclear factor-${\kappa}B$(NF-${\kappa}B$)의 핵 내 전이 pathway를 저해함으로써 나타난 결과인지 확인하기 위하여 iNOS, COX-2 및 NF-${\kappa}B$ p65의 단백질 발현량을 관찰한 결과, 비교적 낮은 농도인 $50{\mu}g/mL$에서 40% 이상의 저해능을 보인 것으로 보아 NO와 cytokine의 분비 억제 결과가 NF-${\kappa}B$ pathway를 저해함으로써 나타난 것임을 유추할 수 있었다. 또한 LPS에 의해 증가한 mitogen-activated protein kinases의 인산화를 확인한 결과, CNYEE 처리에 의해 농도 의존적으로 유의성 있게 저해되었다. 이러한 결과를 종합해볼 때 가락진두발 에탄올 추출물은 염증매개물질의 분비를 효과적으로 저해함으로써 추후 천연물로서 염증 치료제의 개발이 가능할 것으로 생각한다.

• 생약학회지
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• 제51권4호
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• pp.244-254
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• 2020

Atractylodes macrocephala is a perennial herb and is a member of the Compositae family. This plant is known to contain various bioactive constituents indicating anti-inflammatory, neuroprotective, anti-oxidant, immunological enhancement, and gastroprotective effects. In this investigation, we isolated four compounds with similar chemical structures from A. macrocephala, and evaluated their anti-inflammatory effects. Among the four compounds, compound 2(atractylenolide II) showed the second-best inhibitory effect on the lipopolysaccharide(LPS)-induced production of nitric oxide in RAW264.7 macrophages and BV2 microglial cells. Compound 2 also inhibited the LPS-induced the production of prostaglandin E2(PGE2), and the expression of inducible nitric oxide synthase(iNOS) and cyclooxygenase(COX)-2 proteins in both cells. In addition, compound 2 suppressed the production of pro-inflammatory cytokines including interleukin(IL)-1β, IL-6, and tumor necrosis factor(TNF)-α. These inhibitory effects were contributed by inactivation of nuclear factor kappa B(NF-κB) and mitogen-activated protein kinases(MAPKs) pathways by treatment with compound 2. This compound did not induce the expression of heme oxygenase(HO)-1 protein indicating that the anti-inflammatory effect of compound 2 was independent with HO-1 protein. Taken together, these results suggested that atractylenolide II can be a candidate material to treat inflammatory diseases.

• Journal of the Korean Wood Science and Technology
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• 제47권6호
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• pp.674-691
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• 2019

본 연구는 국내산 편백, 소나무, 잣나무, 일본잎갈나무 목부 정유의 항염증 효과를 평가 하였다. 실험에 사용된 정유는 각 수종의 목부에서 증류법(hydrodistillation)으로 추출되었다. 추출한 정유들을 GC-MS를 통해 성분 분석한 결과, 편백의 경우 ${\alpha}$-cadinol (19.25%), ${\tau}$-muurolol (14.20%) 및 ${\alpha}$-pinene (13.74%)이, 소나무는 ${\alpha}$-pinene (47.16%), longifolene (14.31%), ${\beta}$-phellandrene (11.78%) 및 ${\beta}$-pinene (11.02%)이 주요성분이었으며, 잣나무의 경우 ${\alpha}$-pinene (13.49%)와 longifolene(10.79%), 낙엽송의 경우 geranyl linalool (23.58%) 및 ${\alpha}$-pinene (18.57%)이 주요성분으로 확인되었다. 침엽수 목부 정유들의 항염증 효과는 LPS(Lipopolysaccharide)로 염증반응이 유도된 RBL-2H3 비만세포주에 정유를 처리한 후 IL-4(interleukin-4), IL-13(interleukin-13), ${\beta}$-hexosaminidase의 발현 변화를 조사함으로써 평가하였다. MTT assay를 통한 세포 독성 평가 결과, 침엽수 4종 정유 모두 $10^{-7}{\sim}10^{-5}%$ 농도에서 세포 독성을 보이지 않았다. 염증반응이 유도된 RBL-2H3 세포에 침엽수 4종의 목부 정유를 $10^{-7}%$ 농도로 처리하였을 때, LPS 처리군 대비 침엽수 4종의 목부 처리군에서 IL-4와 IL-13의 상대적 발현량이 큰 폭으로 감소하였으며, ${\beta}$-hexosaminidase 분비가 유의하게 억제되었다. 침엽수 정유 4종의 IL-4 상대적 발현량은 LPS 처리군 대비 소나무 69.6%, 잣나무 63.2%, 편백 55.1% 그리고 낙엽송 45.8% 감소하였다. IL-13의 상대적 발현량은 소나무 57.8%, 잣나무 57.1%, 편백 51.1% 그리고 낙엽송 34.5% 감소하였다. ${\beta}$-Hexosaminidase 분비의 경우 LPS 처리군 대비 편백 38.1%, 소나무 33.0%, 잣나무 27.4% 그리고 낙엽송 9.1% 억제하였다. Il-4, IL-13 및 ${\beta}$-hexosaminidase 분비를 비교한 결과, 침엽수 목부 정유 4종 중 소나무 정유의 활성이 가장 우수하였다. 이러한 결과는 국내 침엽수 4종의 목부 정유는 염증반응을 유발하는 싸이토카인의 발현 억제를 통하여 항염증 효과를 나타냄을 확인할 수 있었다.

• 대한한방부인과학회지
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• 제30권2호
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• pp.1-15
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• 2017

Objectives: The object of this study was to identify the anti-inflammatory effects of Patrinia scabiosaefolia aqueous extract (PSE). Methods: RAW 264.7 cells were pre-treated with PSE and then incubated with or without lipopolysaccharide (LPS). Cell viability, production of nitric oxide (NO), secretion of pro-inflammatory cytokine, activation of mitogen-activated protein kinases (MAPKs) and nuclear factor-kappa B (NF-${\kappa}B$) were measured. In addition, we observed mice survival rate after LPS and their cytokine levels of serum. We also observed inflammatory and hemorrhagic change on the histological sections of the liver. Results: PSE inhibited LPS-induced NO production, interleukin (IL)-6 secretion, c-Jun NH2-terminal kinase (JNK) and NF-${\kappa}B$ activation. In addition, PSE reduced the death rate of LPS-induced mice and IL-6 production on the serum of mice. PSE inhibited inflammation and hemorrhage on liver tissue as well. Conclusions: The results suggest that PSE have anti-inflammatory effects by inhibited NF-${\kappa}B$ and JNK activation, IL-6 secretion, and NO production. So PSE may be effective treatment for the inflammatory disease.

• 동의생리병리학회지
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• 제18권6호
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• pp.1622-1627
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• 2004

Nephrite has been widely used as a medicinal mineral resource to treat a numerous chronic diseases and to replenish vital essence and blood in the Korean traditional medicine. However, as of yet, there is little understanding of the pharmacological and biochemical mechanisms of its therapeutic effects as regards anti-inflammation. We therefore examined whether nephrite represses the expression of major inflammation mediators, such as interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2), in LPS-stimulated murine macrophage cell line, RAW264.7 by using RT-PCR. The powder suspension and water extracts of nephrite significantly inhibited the mRNA expression of the mediators, despite a little toxic effects on growth of RAW 264.7 cells within the concentration range tested. These experimental results suggested that the nephrite can be utilized as a functional mineral exerting the anti-inflammation medicinal effect.

• Nutrition Research and Practice
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• 제11권5호
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• pp.357-364
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• 2017

BACKGROUND/OBJECTIVES: Oxidative stress is closely related with inflammation and development of many diseases. Black soybean seed coat contains high amount of anthocyanins, which are well-known for free radical scavenging activities. This study investigated inflammatory response and action mechanism of black soybean anthocyanins with regard to antioxidant activity in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. MATERIALS/METHODS: RAW 264.7 cells were treated with anthocyanins extracted from black soybean seed coats in a concentration range of 12.5 to $100{\mu}g/mL$. The production of reactive oxygen species (ROS), secretion of pro-inflammatory mediators and cytokines, and the signaling in the mitogen activated protein kinases (MAPKs) pathway were examined. RESULTS: Black soybean anthocyanins significantly decreased LPS-stimulated production of ROS, inflammatory mediators such as nitric oxide (NO) and prostaglandin $E_2$, and pro-inflammatory cytokines, including tumor necrosis factor ${\alpha}$ and interleukin-6, in a dose-dependent manner without cytotoxicity (P < 0.001). Black soybean anthocyanins downregulated the expression of inducible NO synthase and cyclooxygenase-2 in LPS-stimulated RAW 264.7 cells (P < 0.001). Moreover, black soybean anthocyanins inhibited LPS-induced phosphorylation of MAPKs, including extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38 (P < 0.001). CONCLUSION: These results suggest that black soybean anthocyanins exert anti-inflammatory activity by inhibiting ROS generation and subsequent MAPKs signaling, thereby inhibiting inflammatory responses.

• ALGAE
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• 제32권3호
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• pp.261-273
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• 2017

Fine dust (FD) particles have become a major contributor to air pollution causing detrimental effects on the respiratory system and skin. Although some studies have investigated the effects of FD on the respiratory system, their possible effects on the skin remain under-explored. We investigated the FD mediated inflammatory responses in keratinocytes, present in the outer layers of skin tissues and the transfer of inflammatory potential to macrophages. We further evaluated the anti-inflammatory effects of the polyphenolic derivative, diphlorethohydroxycarmalol (DPHC) isolated from Ishige okamurae against FD-induced inflammation. Size distribution of FD particles was analyzed by scanning electron microscopy. FD particles induced the production of cyclooxygenase-2, prostaglandin E2 ($PGE_2$), interleukin (IL)-$1{\beta}$, and IL-6 in HaCaT keratinocytes and the expression of nitric oxide (NO), inducible nitric oxide synthases (iNOS), $PGE_2$, tumor necrosis factor-${\alpha}$ expression in RAW 264.7 macrophages. Further, we evaluated the inflammatory potential of the culture medium of inflammation-induced HaCaT cells in RAW 264.7 macrophages and observed a marked increase in the expression of NO, iNOS, $PGE_2$, and proinflammatory cytokines. DPHC treatment markedly attenuated the inflammatory responses, indicating its effectiveness in suppressing a broad range of inflammatory responses. It also showed anti-inflammatory potential in in-vivo experiments using FD-stimulated zebrafish embryos by decreasing NO and reactive oxygen species production, while eventing cell death caused by inflammation.

• Archives of Pharmacal Research
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• 제28권10호
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• pp.1170-1176
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• 2005

2-Methyl-2-(2-methylpropenyl)-2,3-dihydronaphthoquinone[2,3-b]furan-4,9-dione (N FD-37) is a synthetic furonaphthoquinone compound. In this study, we determined that NFD-37 could inhibit the lipopolysaccharide (LPS)-induced production of inflammatory mediators in macrophages RAW 264.7. This compound inhibited LPS-induced nitric oxide (NO) or prostaglandin (PG) $E_{2}$ production in dose-dependent manners, with $IC_{50}$ values of 7.2 ${\mu}M$ and 5.3 ${\mu}m$, respectively. As the positive controls, pyrrolidine dithiocarbamate (30 ${\mu}M$) exhibited a $57{\%}$ inhibition of NO production, and NS-398 ($1{\mu}M$) manifested a $48{\%}$ inhibition of $PGE_2$ production. The inhibitory effects of NFD-37 on NO and $PGE_2$ production were determined to occur in conjunction with the suppression of inducible NO synthase or cyclooxygenase-2 expression. NFD-37 also inhibited the production of LPS-inducible tumor necrosis factor-${\alpha}$, interleukin (IL)-$1{\beta}$ and IL-6, at $IC_{50}$ values of 4.8-8.9 ${\mu}M$. We also determined the anti-inflammatory efficacy of NFD-37 using carrageenin-induced paw edema in experimental mice.

• Journal of Ginseng Research
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• 제30권4호
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• pp.181-187
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• 2006

Protopanaxadiol (PPD) is a mixture of protopanaxadiol type saponins with a dammarane skeleton, from Korean red ginseng (Panax ginseng C.A. Meyer; Araliaceae). Korean ginseng is well-known herb to treat almost all kinds of diseases in Oriental medicine. This herb was particularly prescribed for treatment various inflammatory diseases, including rheumatoid arthritis, atherosclerosis, and diabetes mellitus, for centuries. To understand the efficacy of ginseng against inflammatory diseases, we aimed to show anti-inflammatory activities of the PPD in murine macrophage cell line, RAW264.7 cells using nitric oxide (NO) production assay and the expressions of pro-inflammatory cytokines, such as tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), and IL-6, and monocyte chemotactic protein-1 (MCP-1). We found that PPD saponin significantly blocked LPS ($1{\mu}g/ml$)-induced NO production in a dose-dependent manner. In addition, PPD abrogated the expressions of LPS-induced pro-inflammatory cytokines, such as IL-$1{\beta}$ and MCP-1. Moreover, cyclooxygenase (COX)-2, a critical enzyme to produce prostaglandin E2 (PGE2), was significantly inhibited by PPD in LPS-activated RAW264.7 cells. Taken together, these results suggested that anti-inflammatory efficacy of Korean red ginseng on inflammatory diseases is, at least, due to the NO inhibitory activity and the inhibition of the expressional level of inflammatory cytokines and/or mediators.

• Journal of Veterinary Science
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• 제23권1호
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• pp.7.1-7.14
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• 2022

Background: Enterotoxigenic Escherichia coli (ETEC) infection is a primary cause of livestock diarrhea. Therefore, effective vaccines are needed to reduce the incidence of ETEC infection. Objectives: Our study aimed to develop a multivalent ETEC vaccine targeting major virulence factors of ETEC, including enterotoxins and fimbriae. Methods: SLS (STa-LTB-STb) recombinant enterotoxin and fimbriae proteins (F4, F5, F6, F18, and F41) were prepared to develop a multivalent vaccine. A total of 65 mice were immunized subcutaneously by vaccines and phosphate-buffered saline (PBS). The levels of specific immunoglobulin G (IgG) and pro-inflammatory cytokines were determined at 0, 7, 14 and 21 days post-vaccination (dpv). A challenge test with a lethal dose of ETEC was performed, and the survival rate of the mice in each group was recorded. Feces and intestine washes were collected to measure the concentrations of secretory immunoglobulin A (sIgA). Results: Anti-SLS and anti-fimbriae-specific IgG in serums of antigen-vaccinated mice were significantly higher than those of the control group. Immunization with the SLS enterotoxin and multivalent vaccine increased interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) concentrations. Compared to diarrheal symptoms and 100% death of mice in the control group, mice inoculated with the multivalent vaccine showed an 80% survival rate without any symptom of diarrhea, while SLS and fimbriae vaccinated groups showed 60 and 70% survival rates, respectively. Conclusions: Both SLS and fimbriae proteins can serve as vaccine antigens, and the combination of these two antigens can elicit stronger immune responses. The results suggest that the multivalent vaccine can be successfully used for preventing ETEC in important livestock.