• Title/Summary/Keyword: M1/M2 macrophage

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An Atopic Preventive Drink (APD) reduces Th2 cytokines in LPS-treated RAW 264.7 cells

  • Song, Gyl-Hoon;Park, Eui-Seong;Lee, Seung-Min;Kim, Tae-Young;Park, Kun-Young
    • CELLMED
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    • v.7 no.3
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    • pp.15.1-15.6
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    • 2017
  • We analyzed the effects of an Atopic Preventive Drink (APD) on the regulation of Th2 cytokines using RAW 264.7 macrophage cells. In the evaluation of nitric oxide (NO) production in cells, NO production levels were shown to be elevated only in the APD-treated group in a dose-dependent manner. In the lipopolysaccharide (LPS) with APD-treated group, NO production significantly decreased as APD concentration increased. Further, mRNA expression levels and protein concentrations of pro-inflammatory cytokines in cells were determined. Th2 stimulatory cytokine ($IL-1{\beta}$) and Th2 cytokine (IL-6 and IL-10) levels were significantly reduced in the LPS with APD-treated group compared to the only LPS-treated group. mRNA expression levels of inflammatory-related genes (COX-2 and iNOS) were significantly reduced in the LPS with APD-treated group compared to the only LPS-treated group. These results suggest that APD has an anti-atopic effect by reducing mRNA and proteins expressions of Th2 cytokines and inflammatory-related genes.

Studies on Cosmeceutical Activity of Extracts of Moringa oleifera Extract (모링가 추출물에 대한 화장품약리활성 검증)

  • Kim, So Ra;Yoo, Dan Hee;Yeom, Hyeon Ji;Oh, Min Jeong;Lee, Jin Young
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.3
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    • pp.219-229
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    • 2018
  • The purpose of this study was to investigate the role of the Moringa oleifera (M. oleifera) extract as a cosmetic additive. The tyrosinase and elastase inhibitory effects showed 47% and 39% at $1,000{\mu}g/mL$ concentration, respectively. Also, the collagenase inhibition effect was 31% at $500{\mu}g/mL$ concentration. A cell viability test, measured on macrophage cell (RAW 264.7) and melanoma cell (B16F10) by ethanol extract of M. oleifera, showed 94.2% and 94.8% at $100{\mu}g/mL$ concentration, respectively. In order to confirm anti-inflammatory activity, we examined the inhibitory effects on the production of lipopolysaccharides (LPS)-induced NO in RAW 264.7 cells by Griess assay. As a result, the M. oleifera extract showed a concentration-dependent inhibition of NO production. The protein expression inhibitory effects of M. oleifera extract were measured by western blot at 25, 50, $100{\mu}g/mL$ concentration and the ${\beta}-actin$. Results showed that the expression inhibition rates of the iNOS, COX-2, MITF, TRP-1, TRP-2, tyrosinase protein were decreased by 85.8%, 57.5%, 80.7%, 30%, 29.9%, 23.6% at $100{\mu}g/mL$ concentration, respectively. It was concluded that M. oleifera extracts had the anti-inflammatory and whitening effects and thus could be applied for cosmetics as a natural ingredient.

Lonicerae Flos Inhibited COX-2 and MMP-9 in LPS Induced Arthritis of Mouse through Regulation of MIF (LPS 유도 생쥐 관절염에서 금은화의 MIF 활성 조절이 COX-2와 MMP-9 생성 억제에 미치는 영향)

  • Ahn, Sang-Hyun;Kim, Ho-Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.24 no.2
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    • pp.242-248
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    • 2010
  • The present study was designed in order to determine whether Lonicerae flos (LF) could mitigate rheumatoid arthritis through inhibition of cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 by regulation of macrophage migration inhibitory factor (MIF). We found that MIF mRNA expression in synoviocytes stimulated with phorbol-12-myristate-13-acetate dose-dependantly decreased by LF extract treatment (0.4 - 1.0 mg/$m{\ell}$). The distribution of MIF, COX-2 and MMP-9 positive reacted cells in LPS induced arthritis of mice were decreased by LF (45 mg/kg/day) treatment for 28 days. These data likely indicate that LF may act as MIF inhibitor and may be possible to develop useful agent for rheumatoid arthritis.

Taxifolin Inhibited the Nitric Oxide Production and Expression of Pro-inflammatory Cytokine mRNA in Lipopolysaccharide-stimulated RAW264.7 Cells

  • Rhee, Man-Hee;Endale, Mehari;Kamruzzaman, SM;Lee, Whi-Min;Park, Hwa-Jin;Yoo, Myung-Jo;Cho, Jae-Youl
    • Biomedical Science Letters
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    • v.14 no.3
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    • pp.147-155
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    • 2008
  • In previous works, we found that solvent extract of Opuntia humifusa Raf., a member of the lactaceae family, displayed potent anti-oxidative and anti-inflammatory activities. Thus, all solvent fractions, except for the water layer, showed potent scavenging effects. According to activity-guided fractionation, one of active radical scavenging principles in the ethyl acetate fraction was found to be taxifolin. In this study, we investigated whether taxifolin showed anti-oxidative activity. In addition, taxifolin modulated nitric oxide (NO) release and the expression of pro-inflammatory cytokine mRNA such as interleukin-$1{\beta}$ (IL-$1{\beta}$), IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), and TNF-${\alpha}$. Taxifolin showed potent anti-oxidant activity with the $IC_{50}\;of\;8.5{\pm}1.4\;and\;9.3{\pm}1.0{\mu}M$ using xanthine/xanthine oxidase (XO) assay and 2,2-Diphenyl-lpicrylhydrazyl radical (DPPH) assay, respectively. We next determined the role of taxifolin on the immunomodulating activity using murine macrophage cell line RAW264.7 cells. Taxifolin dose-dependently inhibited NO production in lipopolysaccharide (LPS)-activated RAW264.7. It also significantly blocked the expression of inducible NO synthase (iNOS) mRNA in the LPS-stimulated RAW264.7 cells. In addition, taxifolin potently suppressed the expression of IL-$1{\beta}$, IL-6 and GM-CSF mRNA in LPS-activated RAW264.7 cells, but not that of TNF-${\alpha}$ Moreover, taxifolin significantly inhibited the transcriptional activity of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) and activator protein -1 (AP-1). These results suggest that taxifolin may downregulate inflammatory iNOS, IL-$1{\beta}$, IL-6 and GM-CSF gene expressions through inhibition of NF-K and AP-1 activation in LPS-stimulated RAW264.7 cells.

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The Differential Immunomodulating Effects of Levan and DFA-IV on Macrophage Function

  • Park, Sul-Kyoung;Jang, Ki-Hyo;Kim, Mi-Hyun;Lim, Jung-Dae;Han, Eun-Tek;Jang, Seon-A;Kim, Kyung-Ho;Pyo, Suhk-Neung;Sohn, Eun-Hwa
    • Preventive Nutrition and Food Science
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    • v.13 no.1
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    • pp.1-6
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    • 2008
  • Di-D-fructose-2,6':6,2'-dianhydride (DFA-IV) is a disaccharide consisting of two fructose residues that are prepared from levan by levan fructotransferase. Levan is a homopolysaccharide composed of D-fructofuranosyl residues joined by $\beta$-(2,6) and $\beta$-(2,1) linkages. We compared the immunomodulatory effects of levan with DFA-IV. Tumoricidal activity, phagocytosis and nitric oxide (NO) production were examined in levan- and DFA-IV-treated RAW264.7 cells. The NO production, tumoricidal and phagocytic activities were significantly increased in both treated cells. The results indicate that levan has significantly greater effects on tumoricidal activity than DFA-IV at low concentrations (1 ${\mu}g/mL$) and its effect on NO production shows a similar pattern. These results suggest that tumoricidal activity induced by both samples is mediated by NO production.

Production of TNF-${\alpha}$ and IL-6 in Macrophages by Mycobacterial Protein Antigens (결핵균 단백항원 자극에 의한 대식세포의 TNF-${\alpha}$ 및 IL-6 생성과 ERK 활성화)

  • Ahn, Hae-Jeong;Cho, Sang-Nae;Paik, Tae-Hyun;Lee, Jung-Lim;Choi, In-Hong
    • IMMUNE NETWORK
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    • v.7 no.1
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    • pp.26-30
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    • 2007
  • Background: Mycobacterial antigens released as PIM, LM, LAM, lipoproteins and other cellular factors may contribute to macrophage and dendritic cell activation through pattern recognition receptors such as TLRs. In this study, we assessed cytokine production and ERK activation with stimulation of several major mycobacterial antigens. Methods: Purified mycobacterial antigens (10, 22, 30, 38kDa) and recombinant antigens (6, 16, 19, 38kDa, Ag85A antigen) were studied. The production of cytokines (TNF-${\alpha}$, IL-12, IL-6) was measured by ELISA. The ERK activation was detected by western blotting. The expression of TLR2 or TLR4 was measured by flow cytometry. Results: Among purified antigens only 30kDa antigen induced production of IL-6 or TNF-${\alpha}$ in THP-1 macrophage cells. When THP-1 macrophage cells were treated with 30kDa antigen, phosphorylation of ERK was detected. ERK activation also occurred in TLR2 transfectant HEK293 cells with 30kDa antigen stimulation. Conclusion: 30kDa antigen is one of the major mycobacterial antigens inducing cytokine production and MAP kinases phosphorylation in macrophages.

Antioxidant and anti-inflammatory activities from fruiting body extracts of Lyophyllum decastes

  • Ki Nam Yoon;Tae Soo Lee
    • Journal of Mushroom
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    • v.21 no.3
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    • pp.101-109
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    • 2023
  • Lyophyllum decastes has been used for culinary purpose. The present study was conducted to evaluate the antioxidant and anti-inflammatory effects from methanol, acetone, and hot water extracts of L. decastes fruiting bodies. The acetone and methanol extracts showed the higher 1,1-diphenyl-2-picryl-hydrazy radical scavenging activities than that of the hot water extract at 0.5-2.0 mg/mL and was comparable to the BHT, the positive control. The ferrous ion chelating effects of the mushroom extracts at 0.5-2.0 mg/mL were significantly higher than that of BHT. The reducing power of acetone extract (2.12) was significantly lower than that of BHT (2.73) at 2.0 mg/ mL. The mushroom extracts also showed inhibitory effects on production of nitric oxide (NO), and expression of inducible nitric oxide synthase (iNOS) in lipopolysaccharide-induced murine macrophage cells in a concentration dependent manner. In vivo anti-inflammatory experiment on carrageenan-induced hind-paw edema of rat model, the acetone extract of the mushroom significantly suppressed the carrageenan-induced rat hind paw edema of rats in a dose dependently. The results suggest that the fruiting bodies of Lyophyllum decastes are a good natural resource of antioxidant and anti-inflammation.

Purification and Identification of Cytotoxic Compounds from the Root of Rumex crispus L. (소리쟁이 뿌리로부터 세포독성물질 분리 및 정제)

  • Cho, Yong Beom;Kim, Jae Yeon;Kwon, Nam Woo;Hwang, Bang Yeon;Kim, Jun Gu;Woo, Sun Hee;Lee, Moon Soon
    • Korean Journal of Medicinal Crop Science
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    • v.27 no.3
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    • pp.208-217
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    • 2019
  • Background: In the present study, we identified two cytotoxic compounds from the root of Rumex crispus L. using a bioassay-based method. Methods and Results: Compared with the other fractions, the diethyl ether ($Et_2O$) fraction of R. crispus root extract exhibited the strongest of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging effect [scavenging concentration 50% $(SC_{50})=63.8{\pm}1.47{\mu}g/m{\ell}$], nitric oxide (NO) production inhibitory effect on the mouse macrophage cell line RAW264.7 [inhibitory concentration 50% $(IC_{50})=60.9{\pm}7.52{\mu}g/m{\ell}$] and cytotoxicity effect on the human hepatoma cell line, HepG2 [lethal concentration 50% $(LC_{50})=115.4{\pm}1.86{\mu}g/m{\ell}$]. According to the bioassay-based method, two cytotoxic compounds were purified from the $Et_2O$ fraction by using column chromatography and preparative high performance liquid chromatography (prep-HPLC). These two compounds were identified as parietin and chrysophanol by using nuclear magnetic resonance (NMR) and liquid chromatography quadruple time of flight mass spectrometry (LC-QTOF-MS). In addition, both parietin and chrysophanol exhibited a cytotoxicity effect on HepG2 cells, their $LC_{50}$ values were $169.1{\pm}17.67{\mu}M$ and $111.5{\pm}6.62{\mu}M$, respectively. Conclusions: Parietin and chrysophanol isolated from the $Et_2O$ fraction of the R. crispus root extract showed cytotoxicity in HepG2 cell.

Synergistic Anti-inflammatory Effect of Rosmarinic Acid and Luteolin in Lipopolysaccharide-Stimulated RAW264.7 Macrophage Cells (Rosmarinic acid와 luteolin의 항염증에 대한 상승효과)

  • Cho, Byoung Ok;Yin, Hong Hua;Fang, Chong Zhou;Ha, Hye Ok;Kim, Sang Jun;Jeong, Seung Il;Jang, Seon Il
    • Korean Journal of Food Science and Technology
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    • v.47 no.1
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    • pp.119-125
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    • 2015
  • The aim of this study was to investigate the synergistic anti-inflammatory effect of rosmarinic acid (RA) and luteolin from perilla (Perilla frutescens L.) leaves in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. A combination of RA and luteolin more strongly inhibited the production of nitric oxide (NO), inducible NOS (iNOS), prostaglandin $E_2$ ($PGE_2$), and COX-2 than higher concentrations of RA or luteolin alone in LPS-stimulated RAW264.7 macrophages. The combined RA and luteolin synergistically inhibited the production of pro-inflammatory cytokines, such as tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-6 (IL-6), and interleukin-$1{\beta}$ (IL-$1{\beta}$), in LPS-stimulated RAW264.7 macrophages. Furthermore, combined RA and luteolin more strongly suppressed NF-${\kappa}B$ activation than RA or luteolin alone, by inhibiting the degradation of inhibitor of NF-${\kappa}B(I{\kappa}B)$-${\alpha}$ and nuclear translocation of the p65 subunit of NF-${\kappa}B$ in LPS-stimulated RAW264.7 macrophages. Collectively, these results suggest that RA and luteolin in combination exhibit synergistic effects in suppression of LPS-induced inflammation in RAW264.7 macrophages.

Antioxidant activity and suppression of pro-inflammatory mediator of Corni fructus extracts in activated RAW 264.7 macrophage (산수유(Corni fructus) 분획 추출물의 항산화 활성 및 RAW 264.7 대식세포에서 염증매개물질 억제 효과)

  • Kim, Ye Jin;Son, Dae-Yeul
    • Food Science and Preservation
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    • v.23 no.6
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    • pp.876-882
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    • 2016
  • The antioxidant and anti-inflammatory effects of Corni fructus extracts (CEF, EtOAc extraction; CBF, buthanol extraction; CWF, water extraction) were investigated. The total phenolics of CEF (173.3 mg TAE/g) were significantly higher than those of CWF (26.7 mg TAE/g) and CBF (94.8 mg TAE/g). DPPH and ABTS free radical scavenging activity of CEF (DPPH: $RH_{50}$; $25.1{\mu}g/mL$, ABTS: $RC_{50}$; $36.1{\mu}g/mL$) showed even higher than that of BHA and ${\alpha}-tocopherol$ used as positive control. All three Corni fructus extracts in the concentration of $1{\sim}100{\mu}g/mL$ were effective inhibitors of NO and prostaglandin $E_2$ ($PGE_2$). NO production was inhibited 71.3~92.2% by CEF, 76.8~85.5% by CBF and 74.4~96.9% by CWF, respectively. CEF, CBF and CWF ($1{\sim}100{\mu}g/mL$) inhibited also pro-inflammatory cytokines like $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 very effectively. $TNF-{\alpha}$ was inhibited up to 51.2% by CWF and $IL-1{\beta}$ was inhibited up to 67.1% by CEF. IL-6 was best inhibited by CEF up to 58.9%. This study suggested the potential of Corni fructus for use as an excellent antioxidant substance and inflammatory inhibiting mediators. Therefore CEF, CBF and CWF Corni fructus extracts may be used for therapeutic approach to various inflammatory diseases.