• Animal cells and systems
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• 제12권1호
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• pp.15-21
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• 2008

Nitric oxide(NO) has been known to play important roles in numerous physiologic processes including neurotransmission, vasorelaxation, and cellular apoptosis. Using a mouse cDNA gene chip, we examined expression patterns and time course of NO-dependent genes in mouse macrophage RAW264.7 cells. Genes shown to be upregulated more than two fold or at least at two serial time points were further selected and validated by RT-PCR. Finally, 81 selected genes were classified by function as signaling, apoptosis, inflammation, transcription, translation, ionic homeostasis and metabolism. Among those, genes related with signaling, apoptosis and inflammation, such as guanylate cyclase 1, soluble, alpha3(Gucy1a3); protein kinase C, alpha($Pkc{\alpha}$); lymphocyte protein tyrosine kinase(Lck); BCL2/adenovirus E1B 19 kDa-interacting protein(Bnip3); apoptotic protease activating factor 1(Apaf1); X-linked inhibitor of apoptosis(Xiap); cyclin G1(Ccng1); chemokine(C-C motif) ligand 4(Ccl4); B cell translocation gene 2, anti-proliferative(Btg2); lysozyme 2(Lyz2); secreted phosphoprotein 1(Spp1); heme oxygenase(decycling) 1(Hmox1); CD14 antigen(Cd14); and granulin(Grn) may play important roles in NO-dependent responses in murine macrophages.

• 약학회지
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• 제43권5호
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• pp.635-641
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• 1999

In this study, we investigated the immunopotent activities of lepidan, a water soluble proteoglycan from Lentinus lepideus. To examine whether lepidan may affect nonspecific immune responses, we determined the effect on the production of nitric oxide (NO). Lepidan effectively increased the NO production in IFN-${\gamma}$ and LPS triggered RAW 264.7 cells. To further demonstrate the evidence that lepidan activates various immune cells, we determined the alkaline phosphatase activity, plaque-forming cell number and secretion of interleukine-4 (IL-4) and granulocyte/macrophage-colony stimulating factor (GM-CSF) after lepidan treatment in murine splenocytes. The results showed that lepidan increased alkaline phosphatase activity and the number of plaque-forming cells, which indicates that lepidan can lead B lymphocytes to late stage of differentiation. Also, when the splenocytes were cultured with lepidan for 48 hr, the level of IL-4 and GM-CSF in the supernatant dramatically increased. Taken together, these data suggest that lepidan is a biological response modulator that is able to activate immune responses.

• Biomolecules & Therapeutics
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• 제27권4호
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• pp.381-385
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• 2019

We attempted to examine anti-inflammatory and anti-oxidant effects of 4'-O-${\beta}$-D-glucosyl-5-O-methylvisamminol (GOMV), the first epigenetic inhibitor of histone phosphorylation at Ser10. While GOMV did not affect the viability of murine macrophage RAW 264.7 cells, it significantly suppressed lipopolysaccharide (LPS)-induced generation of prostaglandin $E_2$ ($PGE_2$) and nitric oxide (NO) through transcriptional inhibition of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). GOMV also scavenged free radicals in vitro, increased NF-E2-related factor 2 (NRF2), and activated antioxidant response element (ARE), thereby resulting in the induction of phase II cytoprotective enzymes in human keratinocyte HaCaT cells. Finally, GOMV significantly protected HaCaT cells against 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced oxidative intracellular damages. Together, our results illustrate that GOMV possesses anti-inflammatory and anti-oxidant activity.

• Fisheries and Aquatic Sciences
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• 제14권4호
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• pp.267-274
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• 2011

Bacterial lipopolysaccharide (LPS) induces expression of pro-inflammatory cytokines and enzymes producing nitric oxide (NO) and prostaglandins (PGs) in immune cells. This process is mediated by the activation of nuclear factor kappaB (NF-${\kappa}B$). In this study, we investigated the anti-inflammatory characteristics of Codium fragile ethanolic extract (CFE) mediated by the regulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) using LPS-stimulated murine macrophage RAW 264.7 cells. CFE significantly inhibited LPS-induced NO and $PGE_2$ production in a dose-dependent manner and suppressed the expression of iNOS and COX-2 proteins in LPS-stimulated RAW 264.7 cells with no cytotoxicity. Pro-inflammatory cytokines, such as interleukin (IL)-$1{\beta}$, IL-6, and tumor necrosis factor-${\alpha}$, were significantly reduced by treatment of CFE in LPS-stimulated RAW 264.7 cells. CFE inhibited the promoter activity of (NF)-${\kappa}B$ in LPS-stimulated macrophages. Treatment with CFE suppressed translocation of the NF-${\kappa}B$ p65 subunit by preventing proteolytic degradation of inhibitor of ${\kappa}B-{\alpha}$. These results indicate that the CFE-mediated inhibition of NO and $PGE_2$ production in LPS-stimulated RAW 264.7 cells is mediated through the NF-${\kappa}B$-dependent transcriptional downregulation of iNOS and COX-2, suggesting the potential of CFE as a nutraceutical with anti-inflammatory activity.

• 대한화장품학회지
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• 제32권3호
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• pp.173-180
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• 2006

참깨(Sesamum indicum L.)는 1년생 초본식물로서 볶은 참깨나 참기름은 우리나라 식단에서 아주 중요한 조미용 양념 및 식용유지로서 이용되고 있다. 약효로는 동맥경화, 고혈압 및 노화 예방 등으로 알려져 왔으나, 참깨에 함유되어 있는 어떤 성분이, 이들 약효에 관여하고 있는지의 과학적인 규명이 미흡한 실정이다. 본 연구에서는 천연물 유래의 새로운 노화방지제를 개발하기 위해 참깨의 리그난 화합물(sesamin, sesamol, sesaminol, sesaminol diglucosides (SDG), sesaminol triglucosides (STG))의 항염증 활성을 검색하고, 그 중에서 항염증 효과가 가장 높게 나타난 sesamin을 전처치 방법과 동시처치 방법으로 NO 생성억제효과, 세포독성, inducible nitric oxide synthase (iNOS) 및 cyclooxygenases-2 (COX-2) 발현에 미치는 영향을 탐색하고 작용기작을 규명하고자 하였다. 리그난 화합물의 NO 생성억제효과를 탐색한 결과, sesamol, sesaminol, sesamin에서 NO 생성에 미치는 억제율이 높았으나, sesamol, sesaminol은 세포독성이 나타났다. 하지만 sesamin ($IC_{50}: 64{\mu}M$)은 세포독성은 없으면서 농도의존적으로 NO 생성을 억제시켰으며, iNOS 발현에서도 억제 효과를 보여주었다. 결론적으로 sesamin은 피부노화에 원인 중 하나이며 체내 염증반응의 중요 매개인자인 NO의 생성을 억제하는 항염증 물질로서 이와 같은 연구 결과는 향후 임상에서 항염증과 관련된 새로운 항노화 전략으로 개발하기 위해 기초자료로 응응될 수 있음을 시사한다.

• Food Science and Biotechnology
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• 제16권2호
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• pp.205-211
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• 2007

This study was designed to investigate the suppressive effect of chlorophyll a on nitric oxide (NO) production and intracellular oxidative stress. In addition, chlorophyll a regulation of nuclear factor (NF) ${\kappa}B$ activation and inducible NO synthase (iNOS) expression were explored as potential mechanisms of NO suppression in a lipopolysaccharide (LPS)-stimulated macrophage cell line. RAW 264.7 murine macrophages were preincubated with various concentrations ($0-10\;{\mu}g/ mL$) of chlorophyll a and stimulated with LPS to induce oxidative stress and inflammatory response. Treatment with chlorophyll a reduced the accumulation of thiobarbituric acid-reactive substances (TBARS), enhancing glutathione level and the activities of antioxidative enzymes including superoxide dismutase, catalase, glutathione peroxidase (GSH-px), and glutathione reductase in LPS-stimulated macrophages compared to LPS-only treated cells. NO production was significantly suppressed in a dose-dependent manner (p<0.05) with an $IC_{50}$ of $12.8\;{\mu}g/mL$. Treatment with chlorophyll a suppressed the levels of iNOS protein and its mRNA expression. The specific DNA binding activities of NFkB on nuclear extracts from chlorophyll a treated cells were significantly suppressed in a dose-dependent manner with an $IC_{50}$ of $10.7\;{\mu}g/mL$. Chlorophyll a ameliorates NO production and iNOS expression through the down-regulation of NFkB activity, which may be mediated by attenuated oxidative stress in RAW 264.7 macrophages.

• 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.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.447-454
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• 2023

인간이 살아가는 환경에는 인체에 침입하여 건강한 삶을 영위하는 것을 방해하는 다양한 항원들이 존재하며, 면역 체계는 복잡한 기전을 통하여 이를 인식하고 제거한다. 대식세포는 선천 면역체계에 관연하는 면역세포로 체내 널리 분포하고 있으며, inducible nitric oxide synthase로 유도된 산화질소, cyclooxygenase-2로 유도된 prostaglandin E2 그리고 tumor necrosis factor-alpha 등의 전염증성 사이토카인 같은 다양한 면역 조절 물질을 생산한다. 흰점박이꽃무지유충은 미래 식량 수급 문제에 대한 대안으로 등장한 식용 곤충의 일종으로, 기존 mitogen activated protein kinases 및 nuclear factor-kappa B (NF-κB) 신호전달 경로를 경유하는 RAW264.7 대식세포의 활성화를 통한 면역 조절 효과가 보고되었다. 본 연구에서는 RAW264.7 세포에서 흰점박이꽃무지유충 추출물에 의해 유도된 면역 조절 물질의 발현이 toll-like receptor 4, mitogen activated protein kinases 및 nuclear factor-kappa B 신호전달 경로의 약리학적 억제제에 의해 어떻게 변화되었는지 확인하였다. 그 결과, 흰점박이꽃무지유충 처리에 의해 증가된 면역 조절 물질의 발현이 c-Jun N-terminal kinase (JNK) 억제제 및 NF-κB 억제제 처리에 의해 감소하는 것을 확인하였다. 또한, toll-like receptor 4(TLR4) 억제제 처리에 의해서는 흰점박이꽃무지유충 추출물 처리에 의해 증가된 면역 조절 물질의 발현과 JNK 및 NF-κB의 인산화 감소를 확인하였다. 우리의 이러한 연구는 흰점박이꽃무지유충이 TLR4-JNK/NF-κB 신호전달의 관여에 의해 RAW264.7 세포를 활성화하는 것을 시사한다.

• 사상체질의학회지
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• 제22권3호
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• pp.141-151
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• 2010

1. Objectives: Three herbal formulas (Yuldahanso-tang, Chungsimyonja-tang, and Taeeumjowi-tang) for Taeeumin were applied to investigate the immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses in murine macrophage cell line (RAW 264.7) and ovalbumin (OVA)-immunized mice. 2. Methods: This study was carried out in nitric oxide (NO) synthesis in RAW 264.7 cells and cellular proliferation in mouse splenocytes according to three herbal formulas. C57BL/6 mice were immunized intraperitonially with OVA/aluminium ($100\;{\mu}g/200\;{\mu}g$/mouse) on day 1, 8, and 15. Three herbal formulas were administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, IgG1, and total IgM classes) in plasma were measured. 3. Results: Yuldahanso-tang and Chungsimyonja-tang increased NO synthesis in RAW 264.7 cells. Three herbal formulas significantly enhanced cellular proliferation by LPS and Con A in splenocytes from OVA-immunized mice (p<.001). Three herbal formulas for Taeeumin also significantly enhanced plasma OVA-specific IgG, IgG1, and total IgM levels compared with the OVA/Alum group. 4. Conclusion: These results suggested that three herbal formulas for Taeeumin could be used as stimulator of immune response.

• 대한본초학회지
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• 제26권4호
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• pp.187-194
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• 2011

Objective : Astragali Radix (AR) and Cinnamomi Cortex (CC) are used to enhance immune response in Asian traditional medicine. Immuno-potentiation of the combination of AR and CC were evaluated on the cellular and humoral immune response using murine macrophage cell line (RAW 264.7) and OVA-immunized mice. Methods : This study was designed to investigate the immuno-potentiative effects of AR, CC, and AR with CC on nitric oxide synthesis in RAW 264.7 cells and proliferation and production levels of Intereukin-2 (IL-2) in mouse splenocytes. In addition, we evaluated the plasma-specific antibody responses and splenocyte proliferation on ovalbumin (OVA)-immunized mice treated with herbal extracts. Results : Combination treatment with AR and CC increased nitric oxide synthesis in RAW 264.7 cells and IL-2 level in splenocytes (p<0.001). Combination of AR and CC significantly enhanced the Concanavalin A- (Con A ; T cell mitogen) and lipopolysaccharide-(LPS ; B cell mitogen) induced splenocyte proliferation on the OVA-immunized mice. Combination of AR and CC also significantly enhanced plasma levels of OVA-specific IgG (p<0.01), IgG1 (p<0.05) and total IgM (p<0.01) compared with the OVA-immunized control group. Conclusion : These results suggest that combination of AR and CC could be used as therapeutic profile on activation of immune response.