• Preventive Nutrition and Food Science
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• 제11권4호
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• pp.273-277
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• 2006

Recently, Rhei Rhizoma extracts (RRE) have begun to receive more attention as potential biological response modifiers. In the present study, we studied the antiproliferative effect of RRE on tumor cells and the effect of RRE on macrophage function. A variety of tumor cells and macrophages were treated with RRE at various concentrations. The effect of RRE on cell proliferation was measured by MTT assay and the effect of RRE on the production of nitric oxide (NO) was determined in the macrophage-like cell lines Raw264.7, C6 and peritoneal macrophages (pMQ). RRE inhibited the growth of tumor cells (e.g., B16, HOS). However, the effects of RRE on the production of NO varied with macrophage types. RRE had no effect on C6 cell growth and slightly increased the growth of Raw264.7 cells. In addition, treatment of normal pMQ with RRE enhanced NO production in a concentration-dependent manner, whereas RRE suppressed NO production at $50\;{\mu}g/mL$ in both Raw264.7 and C6 cells. However, RRE suppressed NO production in LPS/IFN-$\gamma$-stimulated C6 cells. Overall, these results suggest that RRE elicits an antiproliferative property and differentially modulates NO production in various macrophages, and have a potential for therapeutic application.

• 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.

• 한국축산식품학회지
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• 제30권5호
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• pp.739-745
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• 2010

The objective of the current study was to compare the abilities of undigested and enzymatically digested bifidobacteria to induce nitric oxide and cytokine release in RAW 264.7 macrophage cells. Nine different Bifidobacterium strains derived from herbivorous animals were digested with pepsin and then pancreatin, and the precipitates and supernatants were acquired via centrifugation. The RAW 264.7 cells were incubated with whole cells, the precipitate, or the supernatant, and the macrophage culture supernatants were analyzed with respect to the induction of nitric oxide and cytokines. Pronounced increases in the production of nitric oxide, interleukin (IL)-$1{\beta}$, IL-6, IL-12, and tumor necrosis factor-$\alpha$ (TNF-$\alpha$) were observed when cultured with whole cells and the precipitates. It is noteworthy that the precipitates in most of the Bifidobacterium strains evidenced a trend toward superior IL-12 release compared with whole cells. The results showed that both whole cells and digested Bifidobacterium sp. are effective at stimulating RAW 264.7 cells to induce the production of nitric oxide and cytokines. The pepsin-pancreatin system used in the current study may be useful in unraveling the mechanism by which ingested lactic acid bacteria modulate the induction of macrophage mediators at the cellular level.

• 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.

• Journal of Microbiology and Biotechnology
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• 제15권4호
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• pp.740-744
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• 2005

Bifidobacterium has been previously shown to potentiate immune function, which was mediated through the stimulation of cytokine production by macrophage. This study was performed to further characterize the effective component of Bifidobacterium by measuring the level of interleukin (IL)-6 cytokine using the RAW 264.7 murine cell line as a macrophage model. RAW 264.7 cells were cultured for 24 h in the presence of whole cells (WCs), cell walls (CWs), and cell-free extracts (CFEs) from various strains of Bifidobacterium and other lactic acid bacteria at various concentrations. The most effective component was different depending on the strains and the concentrations used. When tested with each cell fraction from Bifidobacterium sp. BGN4, heat treatment of the cell fractions lowered the production of IL-6. Synergistic effect was obtained, especially when CWs and CFEs were combined. Sonicated WCs stimulated IL-6 production more than intact WCs. The in vitro approaches employed here should be useful in further characterization of the effects of Bifidobacterium on gastrointestinal and systemic immunity.

• 동의생리병리학회지
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• 제23권4호
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• pp.883-887
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• 2009

The effects of Sacchromyces cerevisiae-Fermented Artemisiae Argi Folium Water extract (AFS) on Nitric oxide production from mouse macrophage Raw 264.7 cells treated with EtOH, gallic acid, Nicotine, Acetaminophen, and Acetaldehyde were investigated through this study. AFS (0, 10, 50, 100, 200, 400 ug/mL) was simultaneously treated with EtOH (100 uM), gallic acid (100 uM), Nicotine (1 mM), Acetaminophen (2 mM), and Acetaldehyde (200 uM). And Nitric oxide production from Raw 264.7 cells was measured by Griess reagent method. AFS restorated the cellular production of Nitric oxide reduced by EtOH, gallic acid, Nicotine, and Acetaminophen in Raw 264.7 cells. AFS could be supposed to have the immuno-modulating activity concerned with macrophage's production of Nitric oxide.

• Biomolecules & Therapeutics
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• 제16권4호
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• pp.370-376
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• 2008

Ginsenoside Rp1 (G-Rp1) is a ginseng saponin derivative with chemopreventive and anti-cancer activities. In this study, we examined the regulatory activity of G-Rp1 on the functional activation of macrophages. G-Rp1 remarkably inhibited TNF-$\alpha$ production, LPS-induced cell cytotoxicity, NO production, ROS generation, and phagocytic uptake from lipopolysacchride (LPS)-activated RAW264.7 cells. According to structural feature study using several G-Rp1 analogs, two carbohydrates (glucose-glucose) at R1 position were observedto be highly effective, compared to other structural derivatives. Although the inhibitory activities of G-Rp1 on macrophage functions were not remarkable, several points that G-Rp1 was known to be safe, and that this compound was orally effective, suggest that G-Rp1 may be beneficial in treating macrophage-mediated immunological diseases.

• 한국식품영양과학회지
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• 제27권2호
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• pp.333-337
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• 1998

This study was carried out to get infomation on the nitric oxide production ability and its formation mechanisms of polysaccharides extracted from Ganoderma lucidum(PSG) by using murine macrophage cell line. The cultured mycelial cells of Ganoderma lucidum were extracted by alkali, and than neutralized by acid. The extract were passed through the column of DEAE cellulose for more purification. The neutral fraction was concentrated and precipitated with 95% ethanol. The precipitate was lyophilized and PSG was obtained. The immunomodulating effects of PSG on macrophage were performed by using murine macrophage cell line ATCC TIB 71 cells with PSG 0.5mg. PSG alone could not induce the production of nitrite, but it had a significant potential effect on nitrite secretion when the cells were primed and triggered with BCG and Interferon(IFN)-${\gamma}$. Also it was prominent by using calcium channel blocker(verapamil) and adenylate cyclase activator(forskolin).

• 동의생리병리학회지
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• 제24권4호
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• pp.586-591
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• 2010

Chemokine and Growth Factor are major mediumtors of immuno-inflammatory pathway. The purpose of this study is to investigate whether productions of Chemokine and Growth Factor in lipopolysaccharide (LPS)-induced mouse macrophage RAW 264.7 cells are modulated by Gallic acid (GA), which is easily founded in tannin-containing natural materials such as red wine, green tea, grape juice, and Corni Fructus. Productions of Chemokine and Growth Factor were analyzed by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$ (multi-analyte profiling beads) technology. At first, cell culture supernatant was obtained after treatment with LPS and GA for 24 hour. Then, the antibody-conjugated beads were added and incubated for 30 minutes. After incubation, 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. Based on fluorescence intensity, concentrations of Chemokine and Growth Factor were determined. The results of the experiment are as follows. GA significantly inhibited the production of interferon-inducible protein (IP)-10, keratinocyte-derived chemokine(KC), and vascular endothelial growth factor (VEGF) in LPS-induced RAW 264.7 cells at the concentration of 25, 50, 100, 200 uM (p<0.05). GA significantly inhibited the production of monocyte chemoattractant protein-1(MCP-1) and macrophage-colony stimulating factor(M-CSF) in LPS-induced RAW 264.7 cells at the concentration of 50, 100, 200 uM (p<0.05). GA diminished the production of granulocyte macrophage-colony stimulating factor (GM-CSF) in LPS-induced RAW 264.7 cells. But GA did not show the inhibitory effect on the production of leukemia inhibitory factor (LIP) and macrophage inflammatory protein (MIP)-2 in LPS-induced RAW 264.7 cells. These results suggest that GA has the immuno-modulating activity related with its inhibitory effects on the production of IP-10, KC, MCP-1, VEGF, and M-CSF in LPS-induced macrophages.

• 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.