• 생명과학회지
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• 제18권6호
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• pp.814-825
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• 2008

아스코크로린(Ascochlorin, ASC)은 Ascochyta viciae로부터 추출된 프레닐페놀 물질로, 혈청 콜레스테롤과 트리글리세라이드 수치를 감소시키고 종양 성장을 억제한다는 연구 결과가 보고되어 있다. 본 논문에서는 아스코크로린이 생리학적 혹은 병리학적인 작용과 염증반응에서 약리학적으로 유도되는 반응을 어떻게 조절하며, 이러한 메커니즘에 대해 이해하기 위해 mouse macrophage Raw264.7 세포에 아스코크로린을 처리하여 이에 대한 프로테옴의 특이적인 발현에 대해 분석하였다. 따라서 본 연구는 LPS를 처리한 mouse macrophage Raw264.7 세포에 아스코크로린을 처리하여 염증과정에 관련된 단백질의 발현 양상을 확인하기 위해 프로테오믹스를 시행하였다. Mouse macrophage RAW264.7 세포에 아스코크로린을 처리한 조건과 무처리한 조건으로 나누어 two-dimensional electrophoresis (2-D SDS-PAGE), matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF-MS)와 bioinformatics 방법으로 아스코크로린을 처리한 mouse macrophage Raw264.6 세포의 프로테옴을 분석하였다. 그 결과 mouse macrophage Raw264.7 세포에 아스코크로린 처리 시 Calreticulin이 4배 감소, ${\beta}-actin$도 4배 감소 그리고 vimentin이 1.5배 감소함을 확인 할 수 있었다. 그러나 rabaptin 아스코크로린 처리에 의해 3배 증가함을 확인 할 수 있었다. 이러한 단백질 발현은 RT-PCR을 수행하여 결과에 대해 재확인 하였으며, 프로테오믹스와 동일한 결과를 얻을 수 있었다. 따라서 본 연구를 통해 LPS 처리에 의해 활성화된 mouse macrophage RAW264.7 세포에 ASC를 처리한 후 이차원 전기영동법을 이용하여, 단백질의 발현 변화 및 양상을 규명하고 단백질 지도를 확립 하였으며, RAW264.7 세포를 이용한 면역세포 모델에서 ASC의 항염증 작용을 중심으로 생리활성 조절기능을 확인 할 수 있었다. 향후 분자 기능 조절 연구와 전 임상 연구를 통해 ASC의 생리활성 조절 기능을 규명한다면 ASC는 항염증 및 항암활성을 갖는 약물로 개발될 것으로 기대된다.

• 대한본초학회지
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• 제23권2호
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• pp.151-157
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• 2008

Objectives : The purpose of this study is to investigate the biological Effect of multi-herbal drug 'KOCO-P1' on mouse macrophage Raw 264.7 cells. Methods : Multi-herbal drug 'KOCO-P1' was composed of Ginseng Radix, Astragali Radix, Polygonati Rhizoma, Liriopis Tuber, and Scrophulariae Radix. Cytotoxicity and cytoprotective activity of K0C0-P1 was verificated by MTT assay. And antioxidative effect of K0C0-P1 against EtOH, Nicotine was inspected by Hydroperoxide assay. Results : K0C0-P1 showed no cytotoxicity on RAW 264.7 cells for 24, 48, 72 hours. KOCO-P1 at 200, 100, and 50 ug/mL reduced the production of H202 in Raw 264.7 cells by EtOH. KOCO-P1 at 50 ug/mL reduced the production of H202 in Raw 264.7 cells by Nicotine. Conclusions : KOCO-P1 could be supposed to have antioxidative effect on macrophage with no cytotoxicity.

• 대한약침학회지
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• 제13권3호
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• pp.63-71
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• 2010

Objectives: Gallic acid (GA) is the major component of tannin which could be easily founded in various natural materials such as green tea, red tea, grape juice, and Corni Fructus. The purpose of this study is to investigate the effect of Gallic acid (GA) on production of interleukin (IL) in mouse macrophage Raw 264.7 cells stimulated by lipopolysaccharide (LPS). Methods: Productions of interleukins were measured by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$ (multi-analyte profiling beads) technology. Firstly, cell culture supernatant was obtained after treatment with LPS and GA for 24 hour. Then, it was incubated with the antibody-conjugated beads for 30 minutes. And detection antibody was added and incubated for 30 minutes. And Strepavidin-conjugated Phycoerythrin (SAPE) was added. After incubation for 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System and concentration of interleukin was determined. Results: The results of the experiment are as follows. 1. GA significantly inhibited the production of IL-3, IL-10, IL-12p40, and IL-17 in LPS-induced mouse macrophage RAW 264.7 cells at the concentration of 25, 50, 100, 200 uM (p<0.05). 2. GA significantly inhibited the production of IL-6 in LPS-induced mouse macrophage RAW 264.7 cells at the concentration of 50, 100, 200 uM (p<0.05). 3. GA diminished the production of some cytokine such as IL-4, IL-5, and IL-13 in LPS-induced mouse macrophage RAW 264.7 cells. 4. GA did not show the inhibitory effect on the production of IL-$1{\alpha}$ and IL-9 in LPS-induced mouse macrophage RAW 264.7 cells. Conclusions: These results suggest that GA has anti-inflammatory activity related with its inhibitory effects on the production of interleukins such as IL-3, IL-10, IL-12p40, IL-17, and IL-6 in LPS-induced macrophages.

• 동의생리병리학회지
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• 제22권4호
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• pp.815-820
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• 2008

Macrophage is the important cell for the immune system. Many of herbal drugs were searched about their immune-modulating activity. The purpose of this study is to investigate the biological effect of water extract from ARTEMISIAE ARGI FOLIUM (WAAF) on mouse macrophage Raw 264.7 cells. ARTEMISIAE ARGI FOLIUM was known to have the antibacterial, immune-enhancing, and anticoagulative properties. Cytotoxicity of WAAF was verified by MTT assay. The intracellular production of hydro peroxide ($H_2O_2$) by WAAF was examined. The productions of nitric oxide (NO) and $TNF-{\alpha}$ from Raw 264.7 cell by WAAF were also examined. WAAF showed no cytotoxicity on RAW 264.7 cells for 3 hours. WAAF increased the production of $H_2O_2$ in Raw 264.7 cells. WAAF decrease the production of NO from the cells at low concentrations but increased at high concentrations. WAAF increased the production of $TNF-{\alpha}$ from the cells. Therefore, It could be suggested that WAAF has the immune-modulating effect.

• 동의생리병리학회지
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• 제36권2호
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• pp.66-72
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• 2022

Flos Lonicerae Japonicae (the flower buds of Lonicera japonica Thunberg) has been used as an antibacterial and antiviral drug in Korean Medicine. The aim of this study is to evaluate the effect of Flos Lonicerae Japonicae water extract (FL) on the production of cytokines in RAW 264.7 mouse macrophages stimulated by lipopolysaccharide (LPS). After 24 h treatment, the production of various cytokines from RAW 264.7 was measured with multiplex cytokine assay using Bio-Plex 200 suspension array system. FL at concentrations of 50, 100, and 200 ㎍/mL significantly inhibited productions of tumor necrosis factor-α, macrophage inflammatory protein (MIP)-1β, and MIP-2 in LPS-stimulated RAW 264.7 cells; FL at concentrations of 100 and 200 ㎍/mL significantly inhibited productions of leukemia inhibitory factor, LIX (CXCL5), and RANTES in LPS-stimulated RAW 264.7 cells; FL at concentrations of 200 ㎍/mL significantly inhibited productions of granulocyte-macrophage colony-stimulating factor and macrophage colony-stimulating factor in LPS-stimulated RAW 264.7 cells; FL at concentrations of 50 and 100 ㎍/mL significantly increased productions of interleukin (IL)-10 in LPS-stimulated RAW 264.7 cells; FL at concentrations of 50, 100, and 200 ㎍/mL significantly increased productions of IL-6 and interferon gamma-induced protein-10 in LPS-stimulated RAW 264.7 cells; FL at concentrations of 100 and 200 ㎍/mL significantly increased productions of monocyte chemoattractant protein-1 in LPS-stimulated RAW 264.7 cells. Taken together, these data mean that FL might modulate productions of cytokines, chemokines, and growth factor in LPS-stimulated macrophages. Further study needs to verify the exact mechanism for modulatory activities of FL with macrophages.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2020년도 춘계학술대회
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• pp.86-86
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• 2020

The leaves of Staphylea bumalda (S. bumalda) as a deciduous tree distributed in Korea, China and Japan are used to treat respiratory diseases or inflammation. However, there is no scientific research on the immune-enhancing activity of S. bumalda leaves. Thus, in this study, we investigated the effect of water extracts from S. bumalda leaves (SBL) on the macrophage activity using mouse macrophage cells, RAW264.7. SBL increased production of immunomodulators such as NO, iNOS, IL-1β, IL-6, TNF-α and MCP-1 in RAW264.7 cells and activated phagocytic activity of RAW264.7 cells. Inhibition of TLR2 and TLR4 blocked SBL-mediated production of immunomodulators in RAW264.7 cells. In addition, SBL-mediated production of immunomodulators was attenuated by JNK inhibition in RAW264.7 cells. SBL increased JNK phosphorylation, while Inhibition of TLR2 and TLR4 blocked SBL-mediated JNK phosphorylation in RAW264.7 cells. These results are thought to be evidence that SBL activates JNK through stimulation of TLR2 and TLR4 in macrophage to induce the production of immunomodulators. In LPS-stimulated RAW264.7 cells, SBL inhibited over-production of immunomodulators. Summarizing the results, SBL showed immunostimulatory activity under normal conditions and immunosuppressive activity under LPS-induced excessive immune response conditions.

• 대한본초학회지
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• 제37권1호
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• pp.41-50
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• 2022

Objectives : The aim of this study was to investigate the effect of water extract of Agastache rugosa (AR) on productions of inflammatory mediators in lipopolysaccharide (LPS)-stimulated RAW 264.7 mouse macrophages. Methods : Cell viabilities were measured with MTT assay. The production of nitric oxide (NO) from RAW 264.7 cells was measured with Griess reagent assay. The production of cytokines in RAW 264.7 cells was measured with multiplex cytokine assay. Results : AR showed no cytotoxicity on RAW 264.7 cells. AR at concentrations of 25, 50, 100, and 200 ㎍/mL significantly inhibited NO production in LPS-stimulated RAW 264.7 cells. AR at concentrations of 50, 100, and 200 ㎍/mL significantly inhibited productions of TNF-α and IL-1β in LPS-stimulated RAW 264.7 cells; AR at concentrations of 50 and 200 ㎍/mL significantly inhibited productions of RANTES (CCL5) in LPS-stimulated RAW 264.7 cells; AR at concentrations of 100 ㎍/mL significantly inhibited productions of macrophage inflammatory protein-1β in LPS-stimulated RAW 264.7 cells; AR at concentrations of 50, 100, and 200 ㎍/mL significantly increased productions of IP-10 (CXCL10) in LPS-stimulated RAW 264.7 cells; AR at concentrations of 100 and 200 ㎍/mL significantly increased MCP-1 (CCL-2) in LPS-stimulated RAW 264.7 cells; AR at concentrations of 50 and 100 ㎍/mL significantly increased productions of IL-10 in LPS-stimulated RAW 264.7 cells. Conclusions : AR might have immunomodulatory effects on productions of NO, cytokines, and chemokines in LPS-stimulated RAW 264.7 mouse macrophages.

• 대한예방한의학회지
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• 제5권1호
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• pp.134-143
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• 2001

The oxidative modification of low density lipoprotein(LDL) has been implicated in the development of atherosclerosis. Oxidized LDL are found in macrophage foam cell, and it can induce an macrophage proliferation in atherosclerotic plaque. In this study, we investigated the hypothesis that gamisoyosan(GS) may reduce atherosclerosis by lowering the oxidiazability of LDL, To achive this goal, we examined the effect of GS on LDL oxidation, nitric oxide production in mouse macrophage cell line, RAW264.7, and the effect of GS on cupuric sulfate-induced cytotoxicity, LDH release, and macrophage activity. GS inhibited the generation of oxidized LDL from native LDL in RAW264.7 cell culture, and decreased the release of LDH from cupric sulfate-stimulated RAW264.7 cell. In other experiments, GS activated RAW264.7 cell, and prolonged the survival time, and increased nitric oxide production in Raw 264.7 cells.

• 대한본초학회지
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• 제25권3호
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• pp.27-34
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• 2010

Objectives : The purpose of this study is to investigate the effect of Fermented Houttuyniae Herba Water Extract (HL) on production of proinflammatory mediators in mouse macrophage RAW 264.7 cells. Methods : Cell viabilities were measured by MTT assay. Effect of HL on nitric oxide (NO) production from RAW 264.7 cells was accessed by Griess reagent assay. Effect of HL on productions of inflammatory cytokines such as interleukine (IL)-17, Interferon $\gamma$-inducible protein (IP)-10, Eotaxin, IL-5, Monocyte Chemotactic Protein-3 (MCP-3), and IL-13 in LPS-induced RAW 264.7 cells was accessed by a multiplex bead array assay based on xMAP technology. Results : The results of the experiment are as follows. 1. Incubation with HL for 24 hours showed significant increase in cell viability of RAW 264.7 mouse macrophages (P < 0.05). 2. HL showed to inhibit NO production from RAW 264.7 cells at the concentrations of 25 and 50 ug/mL significantly (P < 0.05). 3. HL inhibited significantly NO production in LPS-induced RAW 264.7 cells at the concentrations of 25, 50, 100 and 200 ug/mL (P < 0.05). 4. HL inhibited significantly IL-17, IP-10 and Eotaxin in LPS-induced RAW 264.7 cells at the concentrations of 25, 50, 100 and 200 ug/mL (P < 0.05). Conclusions : These results suggest that HL has anti-inflammatory moiety related with its inhibition of NO, IL-17, IP-10, and Eotaxin in macrophages.

• 동의생리병리학회지
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• 제22권6호
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• pp.1495-1499
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• 2008

The purpose of this study is to investigate the effect of water extract from Artemisiae Argi Folium (WAAF) on hydrogen peroxide production within mouse macrophage Raw 264.7 Cells treated with gallic acid, EtOH, LPS, and acetaminophen. WAAF (0${\sim}$400 ug/mL) was treated with gallic acid, EtOH, LPS, acetaminophen. And the intracellular productions of hydrogen peroxide were measured by dihydrorhodamine 123 (DHR) assay. WAAF showed the restoration of the intracellular productions of hydrogen peroxide which were reduced by gallic acid, EtOH, LPS, and acetaminophen in Raw 264.7 Cells. WAAF could be supposed to have the immunological activity related with macrophage's oxidative burst.