• Title/Summary/Keyword: RBL-2H3 mast cell

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A comparative study on the inhibitory effects of mast cell-mediated allergic reactions by artificially cultured and wild Acanthopanax senticosus

  • Yi, Jin-Mu;Jeong, Hyun-Ja;Shim, Kyung-Shik;Lee, Kang-Yong;Kim, Jeong-Sook;Zheng, Cui;Tomoko, Jippo;Lee, Young-Mi
    • Advances in Traditional Medicine
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    • v.1 no.2
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    • pp.21-28
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    • 2000
  • We compared the effect between CAS and WAS(root, stem) on mast cell-mediated allergic reaction. CAS, WAS-root and WAS-stem, significantly inhibited compound 48/80-induced systemic allergic reaction(1g/kg) and histamine release from RPMC(1mg/ml). CAS, WAS-root and WAS-stem also inhibited passive cutaneous anaphylactic reaction. In addition, IgE-induced $TNF-{\alpha}$ secretion from RBL-2H3 was inhibited by pretreatment of CAS, WAS-root or WAS-stem$(0.01{\mu}g/ml)$. Taken together, inhibitory effect on mast cell-mediated allergic reactions of WAS-root is greater than those of WAS-stem but less than those of CAS.

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Effects of Water Extract of Glycyrrhiza uralensis on $\beta$-Hexosaminidase Release and Expression of the Cytokines of RBL-2H3 Mast Cells (감초 추출물이 RBL-2H3 비만세포에서 $\beta$-hexosaminidase 분비 및 Th2 cytokine mRNA 발현에 미치는 효과)

  • Kim, Jeong-Mi;Kim, Dae-Jung;Kim, Tae-Hyeuk;Baek, Jong-Mi;Kim, Hyun-Sook;Choe, Myeon
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.4
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    • pp.231-237
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    • 2010
  • The present study was conducted to investigate the anti-allergic reaction with Glycyrrhiza uralensis. We examined cell viability, $\beta$-hexosaminidase release, IL-4 and IL-13 mRNA expression from RBL-2H3 cell after pre-treatment with 0, 100, 250, 500, 1000${\mu}g/m{\ell}$ of Glycyrrhiza uralensis water extracts. Effects of Glycyrrhiza uralensis on the degranulation and pro-inflammatory cytokines (IL-4 and IL-13) expression were evaluated with $\beta$-hexosaminidase assay, and RT-PCR analysis. We observed that Glycyrrhiza uralensis concentrations from 100${\mu}g/m{\ell}$ to 1000${\mu}g/m{\ell}$ had no effect on cell survival. The release of $\beta$-hexosaminidase decreased significantly with all concentrations of Glycyrrhiza uralensis extracts. The expression of the IL-4 and IL-13 mRNA were decreased by Glycyrrhiza uralensis in dose-dependent manner. These results that Glycyrrhiza uralensis has an anti-histamin effects and controls IL-4, IL-13 secretion on allergic reaction.

Roles of Src-family kinase isoforms, Lyn, Fyn, Fgr, and c-Src on degranulation in RBL-2H3 mast cells (비만세포의 탈과립에 대한 다양한 Src-family kinase의 역할)

  • Lee, Jun-Ho;Mun, Se-Hwan;Ko, Na-Young;Kim, Jie-Wan;Kim, Do-Kyun;Kim, Joo-Dong;Her, Erk;Choi, Wahn-Soo
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.350-355
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    • 2007
  • The rat RBL-2H3 mast cells contain various Src-family kinases. Previous reports with this cell line indicated that Lyn activation is an important initial signaling for the activation of the cells. However, the role and location of other Src-family kinase isoforms which are expressed in the cells are not clear. In this study, we now show that isoforms of Src-family kinases, Lyn, fyn, Fgr, c-Src, and Yes are differentially expressed and located differently in the cells as indicated by RT-PCR, immunoblotting analysis, and confocal microscopy. Lyn and Fgr were located on plasma membrane but on the other hand c-Src and Yes were located on intracellular organelle. All of Src-family kinases were cloned and overexpressed for investigating the roles of the isoforms. Overexpression of Fyn and Fgr, not Lyn and c-Src, stimulated Ag-induced degranulation in the cells. Our findings strongly suggest for the first time that each of Src-family kinase isoform can regulate differentially $Fc{\varepsilon}RI$-mediated signaling in RBL-2H3 mast cells.

Effects of Dopaminergic Drugs on the Mast Cell Degranulation and Nitric Oxide Generation in RAW 264.7 Cells

  • Seol, Il-Woong;Kuo, Na-Youn;Kim, Kyeong-Man
    • Archives of Pharmacal Research
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    • v.27 no.1
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    • pp.94-98
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    • 2004
  • Effects of dopaminergic drugs on the degranulation of mast cells (RBL-2H3 cells) and the nitric oxide production from macrophage cells (RAW 264.7) were studied. Among the dopaminergic agonists and antagonists tested, bromocriptine, 7-OH-DPAT, haloperidol, and clozapine showed potent inhibitions of mast cell degranualtion ($IC_{50} value, 5 \mu$ M). However, these dopaminergic agents did not affect the tyrosine phosphorylations of the signaling components of the high affinity IgE receptor ($Fc\varepsilonRI$), such as Syk, $PLC\gamma1$, and $PLC\gamma2$.; This suggested that these signaling components were not involved in the inhibition of the mast cell degranulation by these compounds. On the other hand, dopamine, bromocriptine, 7-OH-DAPT, and haloperidol markedly inhibited the nitric oxide production from RAW 264.7 cells ($IC_{50}$ values, 10-20$\mu$M). Bromocriptine, a dopamine agonist that is routinely used for the treatment of Parkinsons disease, inhibited the expression of the inducible nitric oxide synthase at an early stage of the LPS-induced protein expression in a dose-dependent manner. The results suggested that these dopaminergic agents, when used for the treatment of dopamine receptors-related diseases, such as Schizophrenia or Parkinsons disease, might have additional beneficial effects.

Cudrania tricuspidata Suppresses Mast Cell-Mediated Allergic Response In Vitro and In Vivo (꾸지뽕나무 추출물의 비만세포 억제에 의한 항알레르기 효과 및 기전)

  • Kim, Young-Mi
    • YAKHAK HOEJI
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    • v.56 no.1
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    • pp.26-34
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    • 2012
  • Mast cells play an important role in early and late phase allergic reactions through allergen and IgE-dependent release of histamine, proteases, prostaglandins, and several multifunctional cytokines. In this study, we investigated whether Cudrania tricuspidata extract (CTE) suppresses IgE-mediated allergic responses in mast cells, an allergic animal model, and its mechanism of action in mast cells. We found that CTE inhibited IgE-mediated degranulation and cytokine production in rat basophilic leukemia (RBL)-2H3 mast cells and bone marrow-derived mast cells (BMMC), as well as passive cutaneous anaphylaxis (PCA) in mice. With regard to its mechanism of action, CTE suppressed the activating phosphorylation of spleen tyrosine kinase (Syk), a key enzyme in mast cell signaling processes and that of LAT, a downstream adaptor molecule of Syk in $Fc{\varepsilon}RI$-mediated signal pathways. CTE also suppressed the activating phosphorylation of mitogen-activated protein (MAP) kinases and Akt. The present results strongly suggest that the anti-allergic activity of CTE is mediated through inhibiting degranulation and allergic cytokine secretion by inhibition of Syk kinase in mast cells. Therefore, CTE may be useful for the treatment of allergic diseases.

Effects of Panax ginseng on Type I Hypersensitivity (제1형 과민 반응에 미치는 고려인삼의 영향)

  • Kim, Young-Ran;Lee, Eun;Lee, Shee-Yong;Kim, Kyeong-Man
    • Journal of Ginseng Research
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    • v.20 no.1
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    • pp.1-6
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    • 1996
  • Effects of Panax ginseng on allergic reactions were studied uslng various in vivo and in vitro experimental models such as 48-hr passive cutaneous anaphylaxis, mediators-induced skin reactions, histamine release from rat peritoneal mast cells, hexosaminidase release from RBL-2H3 cells, and lipoxygenase assay . In all of anti-allergic experiments we conducted, ginseng components (50% ethanol extract or ginseng total saponin or ginsenosides) extracted from Korean red ginseng, did not show significant anti-allergic actions. In 48-hr passive cutaneous anaphylaxis and mediators-induced skin reactions, 50% ethanol extract did not suppress hypersensitivity reactions. Total saponin, 50% ethanol extract, and 8 major ginsenosides did not show inhibitory effects on lipoxygeanse activity. Ginseng total saponin did not inhibit histamine release from rat peritoneal mast cells. All of the ginseng components mentioned above were also tested on RBL-2H3 cells, but none of them inhibited hexosaminidase release from this cell line. These results suggest that Panax ginseng does not have effects on allergic reactions at the level of 50% ethanol extract or total saponin used. All of 8 major saponin components tested ($Rb_1$, $Rb_2$, Rc, Rd, Re, Rf, $Rg_1$, $Rg_2$), did not inhibit lipoxygenase activity and degranulation events.

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The Effects of Prunus Armeniaca Linne Var Fractions on Th2 Cytokine Expression and Atopic Dermatitis of NC/Nga Mouse (행인(杏仁) 분획물이 Th2 cytokine 발현과 NC/Nga mouse의 아토피 피부염에 미치는 영향)

  • Kang, Ki Yeon;Han, Jae Kyung;Kim, Yun Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.30 no.4
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    • pp.29-59
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    • 2016
  • Objectives PRAL (Prunus armeniaca Linne Var) has been known to suppress allergic reaction. However, the cellular target and its mechanism of action were unclear. This study was designed to investigate the effect of PRAL on RBL-2H3 mast cell, which is PMA-Ionomycin-induced activated in vitro and the effect of PRAL on the MNC/Nga mice that are DNCB-induced activated in vivo. Methods In this study, IL-4, IL-13 production were examined by ELISA analysis; IL-4, IL-13, IL-31, IL-31Ra, $TNF-{\alpha}$ and GM-CSF mRNA expression were examined by Real-time PCR; manifestations of AP-1 and MAPKs transcription factors were examined by western blotting in vitro. Then skin rashes have been evaluated and verified the distribution of mast cells by H&E and toluidine blue. Also, WBC, eosinophil and neutrophil, IgE level in serum, $IFN-{\gamma}$, IL-4, IL-5 in the splenocyte culture supernatant, the absolute cell numbers of $CD4^+$, $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$, $CD3^+CD69^+$ in the Axillary Lymph Node (ALN), PBMCs and dorsal skin and IL-5, IL-13, IL-31, IL-31Ra in the dorsal skin by Real-time PCR were all evaluated from the NC/BNga mice. Results As a result of this study, the mRNA expression of IL-4, IL-13, IL-31, IL-31Ra and $TNF-{\alpha}$ and IL-4, IL-13 production, shown in ELISA analysis, were suppressed by PRAL. Results from the western blot analysis showed decrease on the expression of mast-cell-specific transcription factors, including AP-1 and p-JNK, p-ERK. Histological examination showed that infiltration levels of immune cells in the skin of the AD-induced NC/Nga mice were improved by PRAL orally adminstration. Orally- administered PRAL group also showred decreased level of IgE in the serum. This group has shown decreased the level of IL-4, IL-5, but shown elevated $IFN-{\gamma}$ level in the splenocyte culture supernatant. The same group also has shown decreased cell numbers of $CD4^+$, $CD8^+$, $CD3^+CD69^+$ in the ALN, and $CD4^+$, $Gr-1^+CD11b^+$ in the dorsal skin. PRAL oral adminstration increased cell numbers of $CD4^+$, but decreased cell numbers of $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$ in the PBMCs. Conclusions Obtained results suggest that PRAL can regulate molecular mediators and immune cells that are functionally associated with atopic dermatitis (AD) induced in the NC/Nga mice. This may play an important role in recovering AD symptoms and suppressing pruritus.

Inhibition Activity of Plants on IgE-mediated Degranulation of RBL-2H3 Cells

  • Lee, Seung-Eun;Jeong, Hye-Gwang;Lee, Dae-Young;Lee, Jeong-Hoon;Choi, Jehun;Kim, Geum-Soog;Noh, Hyung-Jun;Lee, Jae-Won;Kim, Seung-Yu;Ahn, Young-Sup
    • Korean Journal of Plant Resources
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    • v.28 no.6
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    • pp.718-726
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    • 2015
  • In this study, the inhibitory activities of fifty plant extracts on IgE-mediated degranulation in the rat basophilic leukemia cell line (RBL-2H3 cells) were measured; the release of interleukin (IL)-4 and β-hexosaminidase from IgE-sensitized cells treated with the plant extracts was measured; and the effects of the plant extracts on cell viability were tested. The results of the analysis of plant extracts at 20 μg/ml, including the aerial part of Magnolia sieboldii K. Koch, exhibited suppressive activities upon the release of IL-4. Furthermore, several plant extracts including methanol extracted from Lindera erythrocarpa Makino (aerial part) at the same concentration significantly inhibited the release of β-hexosaminidase. Twenty-six of the plant extracts, including methanol extract of Weigela subsessilis (Nakai) L. H. Bailey (branch), showed a cell proliferation effect of over 80% at 100 μg/ml. In conclusion, the results suggest that the leaf/stem of Geum japonicum Thunb. and the stamen/ovary of Nelumbo nucifera Gaertn., which exhibited effective inhibition on β-hexosaminidase release and IL-4 release from mast cells and showed high cell viability, could be useful candidates as anti-allergy materials.

Carex pumila Extract Supresses Mast Cell Activation and IgE-Mediated Allergic Response in Mice (좀보리사초의 IgE 매개성 알레르기 반응 억제 효과 및 기전)

  • Lim, Hannah;Kim, Young Mi
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.356-362
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    • 2014
  • Allergic diseases have increased rapidly over the past decades, affecting an estimated 20~30% of the population in developed countries. In this study, we investigated whether or not a typical costal sand dune plant Carex pumila (CPE) suppresses the activation of mast cells and IgE-mediated allergic response in vitro and in vivo. As the results, the extract of Carex pumila inhibited antigen-stimulated degranulation in RBL-2H3 cells and Bone marrow-derived mast cells (BMMCs), and IgE-mediated passive cutaneous anaphylaxis (PCA) in mice. CPE also suppressed the production of pro-inflammatory cytokines, TNF-${\alpha}$ and IL-4, in antigen-stimulated mast cells. As its mechanism of action, CPE inhibited the activation of Syk in $Fc{\varepsilon}RI$-mediated signalling pathway, and that of LAT, a downstream adaptor molecule of Syk, in a dose-dependent manner. CPE also suppressed the activation of mitogen-activated protein (MAP) kinases, p38, ERK1/2, JNK, and Akt. Altogether, CPE inhibited mast cell activation and IgE-mediated allergic response by antigen through suppressing the activation of Syk. These results suggest that CPE may be useful for the treatment of allergic diseases.

Anti-Allergic Effect of Hyeongbangjiwang-Tang (형방지황양(荊防地黃揚)의 항Allergy 및 항염증 효과)

  • Nam, Hyun-Wook;Park, Jang-Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.3
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    • pp.567-573
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    • 2009
  • The present study was conducted to investigate the anti-allergic activity of HBT. We investigated the anti-allergic effects of HBT in RBL-2H3 basophilic leukemia cells by compound 48/80, a mast cell degranulator. HBT significantly inhibited ${\beta}$-hexosaminidase and histamine release from compound 48/80 stimulated RBL-2H3 cells. The in vitro anti-inflammatory activities of HBT in LPS-stimulated RAW 264.7 cells were investigated. HBT inhibited NO production in LPS-stimulated RAW 264.7 cells and effectively dowregulated the expression of iNOS mRNA and iNOS protein expression in LPS-stimulated RAW 264.7 cells. These result provide evidences that HBT may be beneficial in the treatment of allergic inflammtory disease.