• 대한본초학회지
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• 제32권3호
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• pp.1-7
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• 2017

Objective : The purpose of this study was to investigate the effects of Scrophulariae Radix Water Extract (SR) on the production of inflammatory mediators in RAW 264.7 mouse macrophages cells induced by lipopolysaccharide (LPS). Method : We examined effect of Scrophulariae Radix Extract on the cell viability of mouse macrophages cells. Futhermore, After 24 hours treatment we investigated anti-inflammatory effect of Scrophulariae Radix Extract by the production of Bio-Plex cytokine assay, concentrations of various cytokines such NO, $interleukin(IL)-1{\alpha}$, IL-3 and interferon inducible protein-10(IP-10). Result : No significant changes have been found in the mouse macrophge cell viability by the Scrophulariae Radix Extract at the concentration of 25, 50, 100 and $200{\mu}g/m{\ell}$. The water extract of Scrophulariae Radix significantly inhibited the production of NO in the LPS-induced macrophage at the concentration of 25, 50, 100 and $200{\mu}g/m{\ell}$. The water extract of Scrophulariae Radix significantly inhibited the production of $IL-1{\alpha}$, IL-3 and IP-10 in the LPS-induced macrophage at the concentration of 50, 100 and $200{\mu}g/m{\ell}$. Conclusion : The water extract of Scrophulariae Radix significantly inhibited the production of NO, $IL-1{\alpha}$, IL-3 and IP-10 at the concentration of $50{\mu}g/m{\ell}$ or higher in the LPS-induced macrophages with no changes in the cell viability of them. These results suggest that water extract of Scrophulariae Radix has anti-inflammatory effect related with its inhibition of proinflammatory cytokines such as $IL-1{\alpha}$, IL-3 and IP-10 in the LPS-induced macrophages.

• 대한한방부인과학회지
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• 제27권4호
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• pp.1-14
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• 2014

Objectives: This study was designed to investigate the anti-tumor metastasis by anti-inflammatory and innate immunomodulating effects of extracts of Jipae-san on cancer cells. Methods: Antimetastatic experiments were conducted in vivo mouse model by using 4T1 mouse mammary carcinoma cells. Cell viability of Jipae-san was tested with 4T1 mouse mammary carcinoma cells, colon 26-M3.1 carcinoma cells and macrophage. In addition expression of $TNF-{\alpha}$ and NO induced by LPS was measured after treating with Jipae-san. To observe innate immunomodulating effects of Jipae-san on macrophage, we measured $TNF-{\alpha}$, IL-12, IL-6 and MCP-1, respectively. Cell cytotoxicity was tested with the macrophage stimulated with Jipae-san and we evaluated the activation of $TNF-{\alpha}$ and NO. And the effect of Jipae-san on metastasis was measured without NK-cell using GM1 serum. Results: Intravenous inoculation of Jipae-san significantly inhibited metastasis of 4T1 mouse mammary carcinoma cells. In an in vitro cytotoxicity analysis, cell growth are closer to 100% less than $1,000{\mu}g/ml$ concentration. The expression of $TNF-{\alpha}$ and NO induced by LPS after treating Jipae-san was down regulated in dose-dependent manner. Level of cytokines such as $TNF-{\alpha}$, IL-12, IL-6 and MCP-1 of Jipae-san group were up regulated in compared to the control group. The macrophage stimulated with Jipae-san significantly inhibits the cancer cell at ratio of 10:1, 20:1. The activation of NO was significantly up regualted in a group of 5:1, 10:1, 20:1. The depletion of NK-cells by anti-asialo GM1 serum partly abolished the inhibitory effect of Jipae-san on tumor metastasis. Conclusions: Jipae-san appears to have considerable activity on the anti-metastasis by inflammation control and activation of innate immune system.

• 대한한의학회지
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• 제34권1호
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• pp.1-14
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• 2013

Objectives: This study was designed to investigate the anti-obesity, anti-diabetic and anti-inflammatory effects of Platycodi radix on obese type 2 diabetes mouse model. Methods: Obese type 2 diabetes mouse model was induced by Surwit's high fat, high sucrose diet for 8 weeks. Models were divided into 4 groups of normal diet (ND, n=10), high fat and high sucrose diet (HFD, n=10), high fat and high sucrose diet with Platycodi radix (PR, n=10), and high fat and high sucrose diet with Metformin (Met, n=10). Body weights were measured every week. After 7 weeks fasting, blood sugar and oral glucose tolerance tests were conducted. After 8 weeks blood samples were taken from mouse hearts and analyzed biochemically. Lipid profile, fructosamine, leptin and weight of epididymal fat pad and liver were measured. Adipose tissue macrophage percentage was analyzed by fluorescence-activated cell sorting (FACS). Results: Compared with the HFD group, body weight, glucose level, fructosamine, weight of epididymal fat pad and adipose tissue macrophage percentage decreased in the PR group. Conclusions: These results suggest that Platycodi Radix has anti-obesity, anti-diabetic, and anti-inflammatory effects on obese type 2 diabetes mouse model.

• 한국식품영양학회지
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• 제25권4호
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• pp.755-761
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• 2012

최근 산삼근의 배양이 실험실에서 가능해짐에 따라 산삼에 대한 많은 연구가 이루어지고 있다. 본 연구에서는 산삼 배양근으로부터 다당을 분리하여 보체계 및 macrophage에 대한 면역자극활성을 측정하였다. 산삼 배양근을 열수 추출한 뒤, 80% ethanol 침전을 통하여 조다당을 추출하여 특성을 검토한 결과, 산삼 배양근 조다당체(WGAR-CP)의 neutral sugar 함량은 64.77%였고, uronic acid 함량은 34.32%이었다. WGAR-CP의 구성당 조성을 확인한 결과, glucose를 높은 비율로 함유하고 있었으며, rhamnose, arabinose, galactose, glucose를 미량 함유하고 있었다. Uronic acid에서는 galacturonic acid를 높은 비율로 함유하고 있었다. 한편, WGAR-CP는 인체 초기 면역반응에 있어 중요한 역할을 담당하는 보체계에 대하여 농도 의존적인 활성을 나타냈다. WGAR-CP를 0.1, 1, 10, 50, $100{\mu}g/m{\ell}$ 농도로 mouse peritoneal macrophage에 처리하여 면역자극을 유도한 후, NO 생성 및 cytokine 분비활성을 평가하였다. 그 결과 WGAR-CP의 NO생성은 저농도에서는 유의적인 활성을 보이지 않았으나, 50, $100{\mu}g/m{\ell}$에서 농도별로 유의적인 차이를 보였다. 또한 50, $100{\mu}g/m{\ell}$ 용량에서 IL-6 생성을 농도 유의적으로 증가시켰고, 10, $50{\mu}g/m{\ell}$에서 농도 유의적으로 IL-12의 생성을 증가시켰다. 이상의 결과를 종합해 볼 때, WGAR-CP는 자연면역계를 구성하고 있는 보체계와 macrophage의 활성인자로 작용할 수 있음이 확인되었고, WGAR-CP에 의해 활성화된 macrophage는 NO 및 IL-6, IL-12와 같은 염증성 cytokine의 분비를 유도함으로써 체내 면역기능을 증강시킬 수 있는 가능성이 있을 것으로 사료되었다.

• IMMUNE NETWORK
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• 제24권2호
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• pp.3.1-3.23
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• 2024

Cigarette smoke extract (CSE)-treated mouse airway epithelial cells (MAECs)-derived exosomes accelerate the progression of chronic obstructive pulmonary disease (COPD) by upregulating triggering receptor expressed on myeloid cells 1 (TREM-1); however, the specific mechanism remains unclear. We aimed to explore the potential mechanisms of CSE-treated MAECs-derived exosomes on M1 macrophage polarization and pyroptosis in COPD. In vitro, exosomes were extracted from CSE-treated MAECs, followed by co-culture with macrophages. In vivo, mice exposed to cigarette smoke (CS) to induce COPD, followed by injection or/and intranasal instillation with oe-TREM-1 lentivirus. Lung function and pathological changes were evaluated. CD68⁺ cell number and the levels of iNOS, TNF-α, IL-1β (M1 macrophage marker), and pyroptosis-related proteins (NOD-like receptor family pyrin domain containing 3, apoptosis-associated speck-like protein containing a caspase-1 recruitment domain, caspase-1, cleaved-caspase-1, gasdermin D [GSDMD], and GSDMD-N) were examined. The expression of maternally expressed gene 3 (MEG3), spleen focus forming virus proviral integration oncogene (SPI1), methyltransferase 3 (METTL3), and TREM-1 was detected and the binding relationships among them were verified. MEG3 increased N6-methyladenosine methylation of TREM-1 by recruiting SPI1 to activate METTL3. Overexpression of TREM-1 or METTL3 negated the alleviative effects of MEG3 inhibition on M1 polarization and pyroptosis. In mice exposed to CS, EXO^-CSE further aggravated lung injury, M1 polarization, and pyroptosis, which were reversed by MEG3 inhibition. TREM-1 overexpression negated the palliative effects of MEG3 inhibition on COPD mouse lung injury. Collectively, CSE-treated MAECs-derived exosomal long non-coding RNA MEG3 may expedite M1 macrophage polarization and pyroptosis in COPD via the SPI1/METTL3/TREM-1 axis.

• Journal of Ginseng Research
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• 제42권1호
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• pp.68-74
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• 2018

Background: Ginsenosides have been reported to have many health benefits, including anti-inflammatory effects, and the resolution of inflammation is now considered to be an active process driven by M2-type macrophages. In order to determine whether ginsenosides modulate macrophage phenotypes to reduce inflammation, 11 ginsenosides were studied with respect to macrophage polarization and the resolution of inflammation. Methods: Mouse peritoneal macrophages were polarized into M1 or M2 phenotypes. Reverse transcription-polymerase chain reaction, Western blotting, and measurement of nitric oxide (NO) and prostaglandin $E_2$ levels were performed in vitro and in a zymosan-induced peritonitis C57BL/6 mouse model. Results: Ginsenoside $Rg_3$ was identified as a proresolving ginseng compound based on the induction of M2 macrophage polarization. Ginsenoside $Rg_3$ not only induced the expression of arginase-1 (a representative M2 marker gene), but also suppressed M1 marker genes, such as inducible NO synthase, and NO levels. The proresolving activity of ginsenoside $Rg_3$ was also observed in vivo in a zymosan-induced peritonitis model. Ginsenoside $Rg_3$ accelerated the resolution process when administered at peak inflammatory response into the peritoneal cavity. Conclusion: These results suggest that ginsenoside $Rg_3$ induces the M2 polarization of macrophages and accelerates the resolution of inflammation. This finding opens a new avenue in ginseng pharmacology.

• 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.

• Journal of Microbiology and Biotechnology
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• 제32권11호
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• pp.1406-1415
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• 2022

The formation of macrophage foam cells stimulated by oxidized low-density lipoprotein (ox-LDL) is deemed an important cause of atherosclerosis. Transcription factor Yin Yang 1 (YY1), which is a universally expressed multifunctional protein, is closely related to cell metabolism disorders such as lipid metabolism, sugar metabolism, and bile acid metabolism. However, whether YY1 is involved in macrophage inflammation and lipid accumulation still remains unknown. After mouse macrophage cell line RAW264.7 cells were induced by ox-LDL, YY1 and proprotein convertase subtilisin/kexin type 9 (PCSK9) expressions were found to be increased while low-density lipoprotein receptor (LDLR) expression was lowly expressed. Subsequently, through reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Western blot analysis, Oil Red O staining and cholesterol quantification, it turned out that silencing of YY1 attenuated the inflammatory response and lipid accumulation in RAW264.7 cells caused by ox-LDL. Moreover, results from the JASPAR database, chromatin immunoprecipitation (ChIP) assay, luciferase reporter assay and Western blot analysis suggested that YY1 activated PCSK9 by binding to PCSK9 promoter and modulated the expression of LDLR in the downstream of PCSK9. In addition, the results of functional experiments demonstrated that the inhibitory effects of YY1 interference on ox-LDL-mediated macrophage inflammation and lipid accumulation were reversed by PCSK9 overexpression. To sum up, YY1 depletion inhibited its activation of PCSK9, thereby reducing cellular inflammatory response, cholesterol homeostasis imbalance, and lipid accumulation caused by ox-LDL.

• KSBB Journal
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• 제15권2호
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• pp.156-161
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• 2000

해양에서 분리한 Streptomyces sp. BH-405의 배양액으로부터 정제하여 얻은 항산화 활성이 우수한 획분 band 2 에 대하여 사람 Low Density Lipoprotein(LDL)의 산화 억제 효과에 대하여 실험하였다. Streptomyces sp. BH-405의 배양액으로부터 분리 정제한 획분 band 2는 LDL에 대한 $5\mu\textrm{m}$ $CuSO_4$ 의 유도 산화를 측정한 결과 100 및 200 $\mu\textrm{g}$/mL에서 LDL의 산화억제 효과가 높았다. 그리고, band 2를 이용한 macrophage 및 J774 유도 LDL의 수식에 대한 항상화 효과도 native LDL에 비하여 높았다. 이때 같은 농도의 band 2를 첨가하여 산화 LDL의 전기영동의 이동거리를 측정한 결과 native LDL보다는 약간 높았으나 Oxid LDL의 대조군보다는 이동거리가 낮으며 공액2중결합의 생성억제 효과도 있었다. 사람 LDL의 산화에 대하여 macrophage 및 내피세포를 이용하여 125I-LDL 산화에 대하여 band 2를 각각 100 및 200 $\mu\textrm{g}$/mL씩 첨가하여 실험한 결과 사람 LDL의 분해는 대조구보다 낮았으며 용량 의존형의 결과를 나타내었다.