• Title/Summary/Keyword: RAW 264.7 Cell Line

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Antiinflammatory and antioxidative effects of Agrimonia pilosa Ledeb

  • Sim, SY;Kim, GJ;Ko, SG
    • Advances in Traditional Medicine
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    • v.7 no.3
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    • pp.217-228
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    • 2007
  • Agrimonia pilosa Ledeb. has long been used for a useful natural agent ameliorating inflammation related symptoms in the folk medicine recipe. This study was performed to investigate effects of Agrimonia pilosa Ledeb.(AP) on the expression of inflammation related genes such as the inducible nitric oxide synthase (iNOS) in macrophage cell line, RAW 264.7 cells. The AP (whole plants) was extracted with 80% ethanol and sequentially partitioned with solvents in order to increase polarity. Among the various solvent extracts of AP, the n-butanol (BuOH) fraction showed the most powerful inhibitory ability against nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW 264.7 cells without affecting cell viability. Reverse transcriptase-polymerase chain reaction and Western blot analysis revealed that the BuOH fraction provided a primary inhibitor of the iNOS protein and mRNA expression in LPS-induced RAW 264.7 cells. The DPPH and OH radical scavenging activities of the several fractions of 80% ethanol extracts of AP significantly increased by EtOAC and BuOH fractions. Thus, the present study suggests that the response of a component of the BuOH fraction to NO generation via iNOS expression provide an important clue to elucidate anti-inflammatory mechanism of AP.

STUDIES ON THE MACROPHAGE INFLAMMATORY $PROTEIN-1{\alpha}$ IN BONE MARROW, SPLEEN, AND MACROPHAGE (비장, 골수세포 및 대식세포에서의 Macrophage Inflammatory $Protein-1{\alpha}(MIP-1{\alpha})$ 에 관한 연구)

  • Song, In-Taeck;Oh, Kwi-Ok;Kim, Hyung-Sup
    • Journal of Periodontal and Implant Science
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    • v.23 no.1
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    • pp.48-55
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    • 1993
  • Macrophage inflammatory $protein-1{\alpha}(MIP-1{\alpha})$ from activated T cell or macrophage, which is small inducible cytokine of unkown biological function, has been shown to display inflammation chemokinetic activities, as well as myelosuppressive effect on more immature progenitor cells. In this paper we show the $MIP-1{\alpha}$ mRNA expression and the presence of $MIP-1{\alpha}$ binding sites from murine macrophage cell line RAW 264.7, and primary cells of mouse bone marrow and spleen. $MIP-1{\alpha}$ mRNA was induced from LPS-stimulated RAW 264.7, but not inhibited by cyclosporin A treatment, and also was expressed from mouse splenocyted and bone marrow cell which were not increased by ferritin or lactoferrin treatment. The results of receptor binding assay showed that radiolabeled RAW 264.7 cell with kd value of 0.91 nM, and binding sites per cell of 378. bone marrow cell and splenocyte also appeared to have $MIP-1{\alpha}$ binding sites 33 and 11 per cell, respectiviely.

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Paraquat Induces Apoptosis through a Mitochondria-Dependent Pathway in RAW264.7 Cells

  • Jang, Yeo Jin;Won, Jong Hoon;Back, Moon Jung;Fu, Zhicheng;Jang, Ji Min;Ha, Hae Chan;Hong, SeungBeom;Chang, Minsun;Kim, Dae Kyong
    • Biomolecules & Therapeutics
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    • v.23 no.5
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    • pp.407-413
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    • 2015
  • Paraquat dichloride (N,N-dimethyl-4-4'-bipiridinium, PQ) is an extremely toxic chemical that is widely used in herbicides. PQ generates reactive oxygen species (ROS) and causes multiple organ failure. In particular, PQ has been reported to be an immunotoxic agrochemical compound. PQ was shown to decrease the number of macrophages in rats and suppress monocyte phagocytic activity in mice. However, the effect of PQ on macrophage cell viability remains unclear. In this study, we evaluated the cytotoxic effect of PQ on the mouse macrophage cell line, RAW264.7 and its possible mechanism of action. RAW264.7 cells were treated with PQ (0, 75, and $150{\mu}M$), and cellular apoptosis, mitochondrial membrane potential (MMP), and intracellular ROS levels were determined. Morphological changes to the cell nucleus and cellular apoptosis were also evaluated by DAPI and Annexin V staining, respectively. In this study, PQ induced apoptotic cell death by dose-dependently decreasing MMP. Additionally, PQ increased the cleaved form of caspase-3, an apoptotic marker. In conclusion, PQ induces apoptosis in RAW264.7 cells through a ROS-mediated mitochondrial pathway. Thus, our study improves our knowledge of PQ-induced toxicity, and may give us a greater understanding of how PQ affects the immune system.

Induction of Nitric Oxide Production by Bafilomycin A1 in Mouse Leukemic Monocyte Cell Line

  • Hong, Jang-Ja;Nakano, Yasuhiro;Ohuchi, Kazuo;Kang, Young-Sook
    • Biomolecules & Therapeutics
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    • v.14 no.3
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    • pp.143-147
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    • 2006
  • In the mouse leukemic monocyte cell line RAW 264.7, the vacuolar-type $(H^+)$-ATPase (V-ATPase) inhibitor bafilomycin $A_1$ at 10 and 100 nM decreased cell growth and survival as determined by 3-(4,5-dimethyl(thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in a concentration-dependent manner. At such concentrations, bafilomycin $A_1$ induced nitric oxide (NO) production through the expression of inducible nitric oxide synthase (iNOS). The bafilomycin $A_1$-induced NO production was inhibited by the NOS inhibitor $N^G$-monomethyl-L-arginine acetate (L-NMMA). Our findings suggest that the V-ATPase inhibitor bafilomycin $A_1$ induces NO production through the expression of iNOS protein.

Anti-oxidant and Anti-inflammatory activities of Zanthoxylum schinifolium Essential Oil (산초 종자 정유의 항산화 및 항염 효능)

  • Kim, Bo-Ae
    • Journal of the Korean Applied Science and Technology
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    • v.31 no.3
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    • pp.440-445
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    • 2014
  • The purpose of this study was to investigate anti-inflammatory and antioxidant effects of essential oil from seed of Zanthoxylum schinifolium on cultured RAW 264.7 cell line. Antioxidant activity of essential oil was evaluated by two different assays as 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and super oxide dismutase (SOD) like activities. This essential oil was tested for cell viability on RAW 264.7 cell line by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. The effects of anti-inflammatory on LPS-induced RAW 264.7 cell line was studied by the content of nitric oxide (NO) and prostagladin $E_2$ ($PGE_2$) in cells. The essential oil of seed from Zanthoxylum schinifolium obtained dose-dependently increased the scavenging activity on DPPH radical scavenging activity and SOD like activity. The essential oil showed low cytotoxicity as more than 98% cell viability in $40{\mu}g/mL^{-1}$ concentration. The essential oil of seed extracted from Zanthoxylum schinifolium presented obvious effect on inflammation. These results suggest that essential oil of seed from Zanthoxylum schinifolium may have value as the potential anti-inflammatory effects by decreasing the action of NO and $PGE_2$ and preventing the activation of oxidative.

Solanum nigrum L. Extract Inhibits Inflammation in Lipopolysaccharide-stimulated Raw 264.7 and BV2 Cells

  • Lee, Jin Wook;Jung, Hyuk-Sang;Sohn, Youngjoo;Kang, Yoon Joong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.92-92
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    • 2018
  • Solanum nigrum L. (SNL), generally known as black nightshade, is traditionally used as medicine to reduce inflammation caused by several diseases like asthma, chronic bronchitis and liver cirrhosis. In this study, anti-inflammatory effects of SNL extract were examined and possible molecular mechanisms of the anti-inflammatory effects were investigated. The inhibitory effects of SNL extract on nitric oxide (NO), pro-inflammatory cytokines ($TNF-{\alpha}$, IL-6) and Matrix metallopeptidase 9 (MMP-9) productions were dissected using lipopolysaccharide (LPS) stimulated murine macrophage-like cell line Raw264.7 cells and human microglial cell line BV2 cells. We further investigated whether SNL extract could suppress the phosphorylation of ERK1/2, JNK, and p38 and the nuclear expression of nuclear factor $NF-{\kappa}B$ p65 in LPS-stimulated Raw264.7 cells and BV2 cells. As a result, we showed that the SNL extract significantly decreased the production of pro-inflammatory cytokines, NO, and MMP-9. In addition, the SNL strongly inhibited the phosphorylation of ERK1/2, JNK, p38 and nuclear translocation of $NF-{\kappa}B$ p65 in activated cells. We confirmed that the extracts of SNL effectively inhibits the anti-inflammatory and may be used as a therapeutic to various inflammatory diseases.

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Effects of Substance P on the Release of Cytokines from Immune Cell Lines (면역세포의 cytokine 유리에 미치는 substance P의 영향)

  • Lee, Jin-Yong;Kim, Soo-Ah;Seo, Seok-Ran;Kim, Hyong-Seop
    • Journal of Periodontal and Implant Science
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    • v.27 no.2
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    • pp.425-441
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    • 1997
  • The neuropeptide substance P(SP) has been implicated in the mediation of inflammation and immune-mediated disease such as arthritis. Recently, it was reported that SP was markedly increased around the blood vessels in inflamed gingiva as well as in close association with the inflammatory cell infiltrate. These results support that SP may contribute to the pathophysiology of neuronal inflammation in human periodontal tissues. SP may regulate inflammatory/immune responses by stimulating the proliferation of human T cells, differentiation and antibody-secreting potential of B cells, macrophage respiratory burst, connective tissue proliferation, and the secretion of cytokines from monocytes and T cells. Here, I studied potential role of SP as a costimulatory chemical signal in inflammatory/immune responses, by determining the released proinflammatory cytokines such as $MIP-1{\alpha}$, $IL-1{\beta}$, and IL-6 from culture supernatants of homogeneous immune cell lines. Serum free cell supernatants were concentrated with TCA precipitation, fractionated with SDS-PAGE, and subjected into western blot analysis. Among 15 cell lines tested, macrophage/monocyte cell line RAW264.7 and WRl9m.1 showed the highest level of induction of $MIP-1{\alpha}$ when stimulated with LPS. Discrete IL-6 bands with multiple forms of molecular mass were detected from supernatants of B cell lines A20(32kDa), Daudi(32, 35kDa), and SKW6.4(29kDa), which were expressed constitutively. $IL-1{\beta}$ could not be detected by the method of western blot analysis from supernatants of all cell lines tested except RAW264.7, WRl9m.1, and erythroid cell line K562 which showed the least amount of $IL-{\beta}$ secretion. SP $10^{-9}M$ with suboptimal dose of LPS treatment showed synergistic induction of $MIP-1{\alpha}$ release from RAW264.7 or WR19m.1, and also IL-6 release from A20, but this synergism is not the case in costimulation of RAW264.7 or WRl9m.1 with SP $10^{-9}M$ and TPA. Although treatment of T cell line CTLL-R8 with SP $10^{-7}M$ or PHA+TPA induced modest level of $MIP-1{\alpha}$ secretion, synergism was not observed when they are applied together. These findings all together suggest the possibility of a regulatory role of SP in inflammatory/immune reaction through differential modulation of bioactivities of other chemical cosignals.

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Anthocyanin Analysis of Pressure-extracted Korean Blueberry Juice and in vitro Anti-inflammatory in RAW267.4 Cell line (국산 블루베리 착즙액의 안토시아닌 분석 및 RAW267.4 세포주에서의 항염효과)

  • Choi, Moon-Hee;Jeon, Young-Jin;Shin, Hyun-Jae
    • KSBB Journal
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    • v.30 no.4
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    • pp.191-196
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    • 2015
  • Blueberry juice possesses rich-procyanidins and - anthocyanidin, comprised a group of with numerous health benefits such as protection against coronary heart disease, detoxification, and obesity. Blueberry (Vaccinium virgatum) juice extracts were analyzed and separated by an HPLC method for the purpose of the separation and quantification in polyphenolic groups. In specific HPLC conditions, a binary mobile phase consisting of formic acid: water (10:90, v/v, solvent A) and formic acid: water: acetonitrile (10:60:30, v/v/v, solvent B) was utilized and it is detected at 546 nm wavelength. The phenolic contents of the extracts are determined using Folin-Ciocalteu phenol reagent. In order to test anti-inflammation activity assay, after producing nitric oxide (NO) in lipopolysaccharide activated RAW 264.7 cells, at concentration of $20-500{\mu}g/mL$ it reduced to NO production at a dose-dependent manner. Importantly, cytotoxicity assay with up to $500{\mu}g/mL$ of the extract from blueberry juice showed ~100% cell viability for RAW264.7 cell line. Therefore, Korean blueberry juice might have potential as anti-oxidant and antiinflammation agents.

Anti-inflammatory Activities of Diarylheptanoid from the Bark of Alnus japonica Steudel

  • Yeom, Seung-Hwan;Kim, Min-Kee;Kim, Hyun-Jung;Shim, Jae-Geul;Lee, Jae-Hee;Lee, Min-Won
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.264.2-264.2
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    • 2003
  • The bark of Alnus japonica has been used of fever, hemorrhage and diarrehea in oriental traditional medicine. This research was focused on the anti-inflammatory activities of diarylheptanoid from the bark of A. japonica on RAW 264.7 cell line. Phytochemical examination of the bark of Alnus japonica Steudel had led to the isolation of ten diarylheptanoids. To investigate the anti-inflammatory activities of these compounds, nitric oxide and PGE2 production inhibitory in IFN-${\gamma}$, LPS stimulated RAW 264.7 cell were examined. (omitted)

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