• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.139.2-139.2
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• 2003

The in vitro effects of extracted fractions of C. militaris on the secretion of cytokines in murine macrophage cell line, RAW 264.7 were studied. F1 (crude cordycepin containing adenosine), F2 (ethanol precipitation), F3 (ethanol soluble supernatant) and F4 (fraction of through SK-1B) significantly stimulated the production of cytokine and nitric oxide (NO) on murine macrophage cell line RAW264.7. We examined how the ethanol extract of C. militaris regulates production of interleukine 1-beta(IL-1$\beta$), tumor necrosis factor-alpha (TNF-$\alpha$), and NO in vitro. (omitted)

• 한국자원식물학회지
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• 제23권5호
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• pp.399-405
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• 2010

현재 임상적으로 적용되고 있는 항암요법들은 많은 부작용을 가지고 있으며, 부작용에 의하여 또다른 질환을 야기하는 것이 항암치료에서 문제점으로 지적되고 있다. 따라서 부작용을 줄이고 항암요법을 유지시키는 방법으로, 인체 안전하다고 보고된 천연물을 이용하여 면역력을 증가시킴으로써 인체내 항암 효과를 나타내게 하는 BRM들을 개발하는 연구가 큰 의미를 지닌다. 이에 본 연구에서는 민간요법으로 이미 사용되고 있는 해당화의 macrophage의 활성화에 대한 BRM 효과를 확인하였으며, 특히 과육(RRF)과 종자(RRS)의 부위별 추출물로 그 효과를 비교 분석하였다. RRF는 암세포 자체에 대한 세포독성은 나타내지 않았으나, macrophage의 활성화에 의한 항암 효과를 나타내었다. 이러한 항암 효과는 macrophage의 활성화에 의한 증가되는 NO 및 TNF-$\alpha$와 같은 암세포 독성물질에 의한 효과를 기대할 수 있으나, RRF 처리에 의하여 활성화된 macrophage는 NO 분비에 효과를 나타내지 않았다. RRF의 처리는 TNF-$\alpha$ 분비를 증가시켰으나, macrophage의 활성화에 의해 암세포 독성을 나타내지 않았던 RRS에서도 TNF-$\alpha$ 분비가 증가한 것으로 보아 TNF-$\alpha$ 분비만으로는 macrophage의 항암효과에 직접적인 영향을 나타내지는 않는 것으로 보인다. 본 연구 결과들은 종합해 볼 때, RRF는 RRS와는 달리 macrophage를 활성화하여 항암효과를 나타내었으며, phagocytosis 능력도 증가시켰다. RRF는 TNF-$\alpha$ 등의 분비조절과 같이 RRS와는 다르게 macrophage를 활성화시키는 것으로 보이며, 임상적으로 적용시 RRF가 RRS보다 세포독성 측면에서도 안전하고, macrophage의 활성화 효과 측면에서도 유의성이 높을 것으로 평가 된다.

• Preventive Nutrition and Food Science
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• 제4권4호
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• pp.265-269
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• 1999

Taurine is a major $\beta$-amino acid in various tissues. Taurine transporter (TAUT) is responsible for the transportation of taurine in the cell. The transporter is affected by various stimuli to maintain its cell volume. Macrophage cell volume varies in its activated states. In our experiment, it was found that the murine macrophage cell line, RAW264.7, expressed TAUT protein in its membrane. Its transportation activities could be blocked by a $\beta$-amino acid such as $\beta$-alanine, but not by $\alpha$-amino acids in this cell line. When assessed in RAW264.7 under the influence of immunosuppressive reagents, the activity of the TAUT was decreased by the treatment of rapamycin (RM) or cyclosporin A (CsA). However when ionomycin (IM) was added to this system, TAUT activity was recovered only in CsA-treated cells in a concentration-dependent manner. In order to inhibit the voltage gated {TEX}$Ca^{+2}${/TEX} channel, calmidazolium was added to the RAW264.7 cell line. Treatment of the cell with calmidazolium completely blocked TAUT. Furthermore, addition of IM to this system recovered the activity of TAUT again. When we added phorbol myristate acetate (PMA) to the cell line, secretion of nitric oxide (NO) was increased 4-fold and the TAUT activity was decreased 5-fold. However, the addition of N-nitro L-arginine methyl ester (L-NAME), an inducible NO synthase (iNOS) inhibitor, to the PMA-treated cells, resulted in the recovery of TAUT activity. These results showed that the activity of TAUT was sensitive to the intracellular concentrations of both {TEX}$Ca^{+2}${/TEX} and NO.

• Journal of Periodontal and Implant Science
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• 제26권2호
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• pp.376-395
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• 1996

The neuropeptide substance P(SP) has been recognized to modulate immune systems, with close proximity between peptidergic sensory nerve endings and immune cells. These include the macrophage and neutrophil activation, IL-2 production in T cell, augmentation of Ig synthesis, mast cell degranulation, $PGE_2$ and collagenase secretion in synoviocytes. In this study I examined SP-induced various biological activities such as antimicrobial action, cytokine production, and mast cell degranulation in the presence or absence of other inflammatory cell activators. Antimicrobial studies showed that undifferentiated HL-60 cells were not affected by SP. However, SP significantly enhanced antimicrobial action of TPA-treated or dbcAMP-treated HL-60 cells which had been differentiated into PMN or macrophage/monocyte. I could not find synergistic relationship between SP and LPS in parallel experiments of the above. SP did not induce IL-l production from murine macrophage cell line RAW264.7 whether costimulated with LPS or not. Mast cell degranulation was occured only when stimulated with high dose ($10^{-5}M$) of SP and the degree of this activation was slightly reduced by simultaneous application of $MIP-1{\alpha}$. In addition, CGRP which is known to be a common coexisting neuropeptide with SP within specific fibers did not augment the function of SP on mast cell degranulation. These results suggest that immunoregulatory activities of SP could be mediated through direct upregulation of various functions of immune cells and also upregulation of responsiveness of immune cells to other immune activators.

• 대한한방부인과학회지
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• 제22권1호
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• pp.41-52
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• 2009

Purpose: This study was designed to investigate the anti-tumor metastasis by immunomodulating effects of extracts of Prunellae Spica. Methods: Antimetastatic experiment was conducted in vivo by using colon 26-M3.1 carcinoma. And we observed cytotoxicity of Prunellae Spica on colon 26-M3.1 carcinoma, L5178Y-R lymphoma cell, hela cell and macrophage. To observe the immnomodulating effects of Prunellae Spica, we estimated IL-6, IL-10, IL-12, TNF-${\alpha}$ from peritoneal macrophages. And we evaluated the activation of NK cell by using anti-asialo-GM1 serum. Results: We found that the administration of Prunellae Spica extracts significantly inhibited tumor metastasis in vivo. In an in vitro cytotoxicity analysis, cell growth are closer to 100% in case of colon 26-M3.1 carcinoma, L5178Y-R lymphoma cell, hela cell at low concentration. In case of macrophage, cell proliferation is closer to 100% less than $62.5{\mu}g/m{\ell}$ of Prunellae Spica extracts. The level of cytokine such as IL-6, IL-10, IL-12 which stimulates Prunellae Spica extracts was increased in dose-dependent manner compared to the control group. TNF-${\alpha}$ is hardly secreted less than $250{\mu}g/m{\ell}$ The depletion of NK cells by anti-asialo GM1 serum partly abolished the inhibitory effect of Prunellae Spica on tumor metastasis. Conclusion: Prunellae Spica appears to have considerable activity on the anti-metastasis by activation the immune system such as macrophage and NK cell.

• Journal of Acupuncture Research
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• 제27권4호
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• pp.223-231
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• 2010

Objectives : The objective of this study is to study the effects of Euonymi Lignum Suberatatum pharmacopuncture solution on NO and IL-6 production in macrophage. Methods : At first, the RAW 264.7 macrophage was subclutured. In order to evaluate cytotoxicity, MTT assay performed. Then, the cell was induced by LPS, INF-$\gamma$ and Experimental groups were divided into five(Normal, Control, Euonymi Lignum Suberatatum 100, 200, $300{\mu}g/m{\ell}$). Then Euonymi Lignum Suberatatum pharmacopuncture solution was put into cell. We measured IL-6, iNOS, NO. Results : The cytotoxic effect of Euonymi Lignum Suberatatum pharmacopuncture solution in RAW 264.7 macrophage was not appeared. $300{\mu}g/m\ell$ Euonymi Lignum Suberatatum pharmacopuncture solution inhibited IL-6 production in LPS, INF-$\gamma$-stimulated RAW 264.7 macrophages significantly. Euonymi Lignum Suberatatum pharmacopuncture solution inhibited iNOS revelation in LPS, INF-$\gamma$-stimulated RAW 264.7 macrophages. All group of Euonymi Lignum Suberatatum pharmacopuncture solution inhibited NO production in LPS, INF-$\gamma$-stimulated RAW 264.7 macrophages significantly. Conclusions : Our study demonstrated that Euonymi Lignum Suberatatum pharmacopuncture solution had an inhibition effect on NO production, iNOS revelation, IL-6 production. So Euonymi Lignum Suberatatum pharmaco puncture solution may have an Anti-inflammation effect.

• 한국가시화정보학회지
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• 제15권3호
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• pp.41-46
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• 2017

The apoptosis of macrophages occurs throughout all stages of atherosclerosis. It is known to constitute atheromatous plaque, increase plaque instability, and thus contribute to the development of atherosclerosis. However, there still remains much to be elucidated on how the apoptotic macrophages affect the endothelial cells and also how they contribute to the development of atherosclerosis. Here we present a microfluidic system, which enables co-culture of apoptotic macrophages and endothelial cells in fibrin gel that mimics in vivo extracellular matrix. With the system, we can investigate the effect of macrophage apoptosis on vascular endothelial cells by quantitatively analyzing the level of reactive oxygen species of HUVECs, integrity of VE-cadherin and cell proliferation. We expect that this system could be utilized further for understanding different mechanisms of apoptotic macrophage on the development of atherosclerosis.

• 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.

• Archives of Pharmacal Research
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• 제22권2호
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• pp.130-136
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• 1999

Mouse guanylate-binding protein 3 (mGBP3) is a 71-kDa GTPase which belongs to GTP-binding protein family. The present study showed that the expression of mGBP3 transcript was readily induced in a dose dependent fashion in the macrophage cell line RAW264.7 treated with either $interferon-{\gamma} (IFN-\gamma)$ or lipopolysaccaride (LPS). The expression of mGBP3 protein was also apparent by 4 and 6 h after the treatment of cells with IFN-\gamma (100 U/ml) or LPS ($1{\mu}g/ml$) , and remained at palteau for at least 24 h. Cycloheximide ($10{\mu}g/ml$) had no effect on the $IFN-\gamma-$ or LPS-induced mGBP3 expression, suggesting that the mGBP3 induction did not require further protein synthesis. Interestingly, a protein kinase C (PKC) inhibitor staurosporine (50 nM) abolished the induction of mGBP3 expression by LPS, but not by $IFN-{\gamma}$. These findings suggest that mGBP3 may be involved in the macrophage activation process and both IFN-\gamma and LS induce the mGBP3 expression through distinct signal transduction pathways.

• 대한본초학회지
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• 제29권1호
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• pp.1-6
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• 2014

Objectives : Many of currently used anti-cancer drugs were developed to target cell death mechanisms and had serious side effects by causing damage to normal cells. Hangambujeongsan or Kangai Fuzheng Powder was a mixture based on the traditional Chinese medicine. It had been used in the local Chinese hospitals to treat cancer patients for decades and had shown a certain level of beneficial effects without major toxic effects. But its mechanism of action had not been elucidated yet. Thus this study aimed to investigate the effects of Kangai Fuzheng Powder in an in vitro experiment. Methods : Cancer lines or RAW264.7 mouse macrophage cells were treated with Kangai Fuzheng Powder. Cell viability was measured by MTT assay, and morphological observation was also performed. Gene expression of cytokines in macrophages was determined by real-time polymerase chain reaction. Phagocytic function assay was also performed in macrophage cells. Results : Kangai Fuzheng Powder had no direct detrimental effect on cancer cells. When macrophages were co-cultured with cancer cells, Kangai Fuzheng Powder had toxic effect on cancer cells. After exposing macrophages to Kangai Fuzheng Powder, macrophages transformed into activated form and the mRNA level of tumor necrosis factor-alpha, interleukin-1beta, interleukin-6, interleukin-10 and monocyte chemotactic protein-1 was significantly enhanced. Phagocytic activity of macrophages was dramatically potentiated. Conclusions : We demonstrated that anti-cancer effect of Kangai Fuzheng Powder was related to activation of macrophages including enhanced cytokine production and phagocytic function.