• 제목/요약/키워드: ${\beta}-Hexosaminidase$ release

검색결과 91건 처리시간 0.026초

RBL-2H3세포에서 생지황약침액의 FcεRI 신호전달을 통한 β-hexosaminidase분비와 Cytokine생성 억제 효과 (Inhibitory Effect of Rehmannia Glutinosa Pharmacopuncture Solution on β-hexosaminidase Release and Cytokine Production via FcεRI signaling in RBL-2H3 Cells)

  • Kang, Kyung-Hwa;Kim, Cheol-Hong
    • 대한약침학회지
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    • 제14권2호
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    • pp.15-24
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    • 2011
  • Background: Type I allergy is involved in allergic asthma, allergic rhinitis, and atopic dermatitis which are accompanied by an acute and chronic allergic inflammatory responses. Rehmannia glutinosa is a traditional medicine in the East Asian region. This study examined whether a Rehmannia Glutinosa pharmacopuncture solution (RGPS) had anti-allergic or anti-inflammatory effects in antigen-stimulated-RBL-2H3 cells. Methods: We determined the effect of RGPS on cell viability using the 3-[4,5-dimethylthiazolyl]-2,5-diphenyltetrazolium bromide (MTT) assay. We also examined the effect of RGPS on the release of ${\beta}$-hexosaminidase and the secretion of IL-4 and TNF-${\alpha}$ using ELISA. In addition, we evaluated the effect of RGPS on the mRNA expression of various cytokines; IL-2, IL-3, IL-4, IL-5, IL-13 and TNF-${\alpha}$ using RT-PCR. Furthermore, we assessed the activation of mitogen-activated protein kinases (MAPKs) and nuclear factor (NF)-${\kappa}$B using Western blotting after RGPS treatment. Results: We found that RGPS ($10^{-4}$ to $10^{-1}$ dilution) did not cause any cytotoxicity. We observed significant inhibition of ${\beta}$-hexosaminidase release and suppression of the protein secretion of IL-4 and TNF-${\alpha}$ and mRNA expression of multiple cytokines in antigen-stimulated-RBL-2H3 cells after RGPS treatment. Additionally, RGPS suppressed not only the phosphorylation of MAPKs, but also the transcriptional activation of NF-${\kappa}$B in antigen-stimulated-RBL-2H3 cells. Conclusions: These results suggest that RGPS inhibits degranulation and expression of cytokines including IL-4 and TNF-${\alpha}$ via down-regulation of MAPKs and NF-${\kappa}$B activation in antigen-stimulated-RBL-2H3 cells. In conclusion, RGPS may have beneficial effects in the exerting anti-allergic or anti-inflammatory activities.

유근피 추출물이 RBL-2H3 비만세포에서 ${\beta}$-hexosaminidase 및 cytokine 분비에 미치는 효과 (Inhibitory effect of Ulmus davidiana on ${\beta}$-hexosaminidase release and cytokine production in RBL-2H3 cells)

  • 박세봉;강경화;윤화정;고우신
    • 한방안이비인후피부과학회지
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    • 제24권1호
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    • pp.86-95
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    • 2011
  • Objectives : Ulmus davidiana (UD) has been widely used in Korean herbal medicines used for treatment of acute and chronic inflammatory diseases, such as rhinitis, asthma, and abscess. In this study, To investigated the protective effect of UD on type 1 allergic response, we determined whether UD inhibits early and late allergic response. Methods : The effect of UD was analyzed by ELISA and RT-PCR in RBL-2H3 cells. Levels of ${\beta}$ -hexosaminidase, interleukin (IL)-4 and TNF-${\alpha}$ were measured using enzyme-linked immunosorbent assays (ELISAs). mRNA levels of COX-2 and T-helper type 2(Th2) cytokines were analyzed with RT-PCR. Results : We found that UD suppressed ${\beta}$-hexosaminidase release in RBL-2H3 not only by the PMA plus A23187 stimulation, but also by the IgE-DNP-HSA stimulation at the antigen-antibody binding stage and antibody-receptor binding stage. UD also significantly inhibited COX2 level, along with reduced Th2 cytokine levels, such as IL-3, IL-4, IL-5, IL-13, GM-CSF, and TNF-${\alpha}$ in RBL-2H3. Conclusions : Our results indicate that UD protects against type 1 allergic response and exerts an anti-inflammatory effect through the inhibition of degranulation and expression of COX2 and Th2 cytokines.

발효처리한 당귀의 항알레르기 효능에 대한 연구 (Anti-allergic Effect of the Fermented Angelicae Gigantis Radix in Human Mast Cell Line HMC-1)

  • 서민준;박진한;이제현
    • 대한본초학회지
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    • 제28권5호
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    • pp.39-44
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    • 2013
  • Objectives : Allergy is an immune dysfunction caused by degranulation from mast cells in the early phase of allergic disease. The purpose of this study was to investigate the anti-allergic effect of fermented Angelicae gigantis Radix in human mast cell line, HMC-1. Method : The Angelicae gigantis Radix was fermented by Lactobacillus acidophilus. The cell toxicity of fermented Angelicae gigantis Radix(FAGR) was determined by MTT assay. The release of ${\beta}$-hexosaminidase from HMC-1 stimulated by phorbol-12-myristate 13-acetate (PMA) plus A23187 was determined by ${\beta}$-hexosaminidase assay. Also, the concentrations of cytokines (interleukin-$1{\beta}$, -6, -8 and tumor necrosis factor-alpha) were measured by enzyme-linked immunosorbent assay. The gene expression of COX-2 from HMC-1 stimulated by phorbol-12-myristate 13-acetate (PMA) plus A23187 was determined by reverse transcription polymerase chain reaction. The release of histamine on substance P-stimulated HMC-1 was measured by histamine assay. Result : The FAGR suppressed the release of ${\beta}$-hexosaminidase, a marker of degranulation, from HMC-1 stimulated by PMA plus A23187. The FAGR inhibited the production of interleukin-$1{\beta}$, -6, -8 and tumor necrosis factor-alpha. The FAGR inhibited the expression of COX-2 mRNA. The FAGR suppressed the release of histamine on substance P-stimulated HMC-1. Conclusion : These results provide that FAGR may be beneficial in the treatment of allergic inflammatory disease.

Suppressive Effects of Benincasae hispida on Allergic Inflammation

  • Park, Seul-Ki;Kim, Jum-Ji;Sung, Sang-Min;Lee, Mi-Young
    • Molecular & Cellular Toxicology
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    • 제5권4호
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    • pp.304-309
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    • 2009
  • The effects of an extract of Benincasae hispida on allergic inflammation were examined in terms of histamine and $\beta$-hexosaminidase release, serum IgE level and inflammatory cytokine level. The B. hispida extract inhibited the release of histamine and $\beta$-hexosaminidase, a degranulation marker, from rat basophilic leukemia cells (RBL-2H3). When mice were first ovalbumin-challenged and then treated with B. hispida extract, there was a significant decrease in the IgE level in the mouse serum. The extract treatment reduced the serum IgE level prominently, compared with the ovalbumin-challenged mice. The extract also significantly reduced the TNF-$\alpha$ and IL-4 levels in the BAL fluid when challenged with antigen. Taken together, the Benincasae hispida extract may be efficacious against allergic inflammation.

Anti-allergic Actions of the Leaves of Castnea crenata and Isolation of an Active Component Responsible for the Inhibition of Mast Cell Degranulation

  • Lee, Eun;Choi, Eun-Ju;Cheong, Ho;Kim, Young-Ran;Ryu, Shi-Yong;Kim, Kyeong-Man
    • Archives of Pharmacal Research
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    • 제22권3호
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    • pp.320-323
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    • 1999
  • The anti-allergic actions of the leaves of Castanea crenata (Fagaceae) were studied. The water extract demonstrated potent anti-allergic actions in in vivo and in vitro experiments. The oral or intraperitoneal administration of the extract (100 or 200 mg/kg) caused a significant inhibition of the 48 hr-PCA (up to 90%) and the vascular permeability induced by histamine or serotonin in rats (about 80%). The anaphylactic release of ${\beta}$-hexosaminidase for RBL-2H3 cells was also significantly inhibited by the extract in as dose-dependent manner with an IC50 value of 230 $\mu\textrm{g}$/ml. The activity-guided fractionation of the extract, based on the determination of inhibitor effect upon the release of ${\beta}$-hexosaminidase, led to the isolation of quercetin as an active principle responsible for the inhibition of degranulation.

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현삼(玄蔘) 추출물이 RBL-2H3 비만세포에서 β-hexosaminidase 및 cytokine 분비에 미치는 효과 (Inhibitory Effects of Scrophulariae Radix on β-hexosaminidase release and cytokine production in RBL-2H3 cells)

  • 김세기
    • 대한본초학회지
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    • 제32권6호
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    • pp.9-15
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    • 2017
  • Objectives : Traditional medicines isolated from natural products often have positive effects in the prevention and healing of various immune disorders, such as allergy and atopic inflammation. Scrophulariae Radix (SR) been used in oriental medicine used for treatment of acute and chronic inflammatory diseases. Mast cells are known to play important roles in the initiation of allergic reactions. In this study, we investigated the effects of SR ethanol extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells. Methods : Rat basophilic leukemia RBL-2H3 cells were purchased from Korean Cell Line Bank (KCLB No. 22256). Cell viability was measured by MTT assay. Assays for ${\beta}-Hexosaminidase$ Secretion : RBL-2H3 cells were sensitized with dinitrophenyl-ImmunoglobulinE (DNP IgE). The next antigen DNP-BSA ($25ng/m{\ell}$) was added for 10 minutes and the reaction was terminated after 5 minutes in the ice bath. To determine ${\beta}-Hexosaminidase$ release, supernatants were aliquoted into 96-well plates. Samples were mixed with substrate solution and incubated for 1 h at $37^{\circ}C$. Absorbance was measured with a spectrophotometer at 405 nm. IL-4 and tumor necrosis $factor-{\alpha}$($TNF-{\alpha}$) concentrations in cell culture supernatants were measured using enzyme-linked immunosorbent assay (ELISA) kits. Results : The cytotoxicity of SRE in RBL-2H3 cells was less than 5%. SRE inhibited DNP-IgE-imduced degranulation of mast cells in RBL-2H3 cells. Also significantly decreased the levels of inflammatory cytokine, IL-4 and TNF-alpha. In this study, the SRE showed potential anti-allergic and antiinflammatory. Conclusions : These results indicate that SRE could be inhibit the allergic response through suppressing the mast cell activation.

만병초와 구상나무 추출물의 RBL-2H3 세포 탈과립, 싸이토카인 유전자 발현에 미치는 영향 (Inhibitory Effect of Extracts from Rhododendron Brachycarpum and Abies Koreana E.H. Wilson on Degranulation and Cytokine Expression in RBL-2H3 Cells)

  • 정의만;김재우;박미진;이성숙;최돈하;이정복;정의배
    • Journal of the Korean Wood Science and Technology
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    • 제41권6호
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    • pp.551-558
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    • 2013
  • 본 연구는 만병초와 구상나무 추출물의 항알러지 효과에 대해 조사하였다. 비만세포 RBL-2H3에 concanavalin A (Con A)를 처리하여 면역반응을 유도한 후 만병초와 구상나무 추출물을 농도별로 처리하여 real-time PCR을 이용하여 IL-4, IL-13의 mRNA 발현량을 확인하였다. 또한 ${\beta}$-hexosaminidase 분비를 측정하여 탈과립 유도효과를 평가하였다. IL-4의 발현은 만병초 $10^{-7}$, $10^{-5}$, $10^{-3}%$ 농도와 구상나무의 모든 농도에서 유의성 있게 감소하였으며, IL-13의 발현은 만병초와 구상나무 모든 농도에서 유의성 있게 감소되었다. 만병초 추출물과 구상나무 추출물의 $10^{-5}$, $10^{-3}%$ 농도에서는 ${\beta}$-hexosaminidase 분비가 유의성 있게 감소하였다. 이러한 결과를 통해 만병초와 구상나무 추출물이 면역반응 표적유전자의 발현을 감소시키고 면역반응에 의해 유도되는 탈과립유도 효과도 감소시킴을 확인할 수 있었다. 이는 만병초와 구상나무 추출물이 항알러지 효과가 있음을 제시하는 결과이다.

The Rhizomes of Acorus gramineus and the Constituents Inhibit Allergic Response In vitro and In vivo

  • Lim, Hyun;Lee, Seung-Young;Lee, Kang-Ro;Kim, Yeong-Shik;Kim, Hyun-Pyo
    • Biomolecules & Therapeutics
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    • 제20권5호
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    • pp.477-481
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    • 2012
  • The rhizomes of Acorus gramineus have frequently been used in traditional medicine mainly for sedation as well as enhancing brain function. In this study, the anti-allergic activity of A. gramineus was investigated. The 70% ethanol extract of the rhizomes of A. gramineus was found to inhibit the allergic response against 5-lipoxygenase (5-LOX)-catalyzed leukotriene (LT) production from rat basophilic leukemia (RBL)-1 cells and ${\beta}$-hexosaminidase release from RBL-2H3 cells with $IC_{50}$'s of 48.9 and > $200{\mu}g/ml$, respectively. Among the 9 major constituents isolated, ${\beta}$-asarone, (2R,3R,4S,5S)-2,4-dimethyl-1,3-bis (2',4',5'-trimethoxyphenyl)tetrahydrofuran (AF) and 2,3-dihydro-4,5,7-trimethoxy-1-ethyl-2-methyl-3-(2,4,5-trimethoxyphenyl)indene (AI) strongly inhibited 5-LOX-catalyzed LT production in A23187-treated RBL-1 cells, AI being the most potent ($IC_{50}=6.7{\mu}M$). Against ${\beta}$-hexosaminidase release by antigen-stimulated RBL-2H3 cells, only AI exhibited strong inhibition ($IC_{50}=7.3{\mu}M$) while ${\beta}$-asarone and AF showed 26.0% and 39.9% inhibition at $50{\mu}M$, respectively. In addition, the ethanol extract of A. gramineus showed significant inhibitory action against the hapten-induced delayed hypersensitivity reaction in mice by oral administration at 200 mg/kg. Therefore, it is suggested that A. gramineus possesses anti-allergic activity and the constituents including ${\beta}$-asarone and AI certainly contribute to the anti-allergic activity of the rhizomes of A. gramineus.

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

  • 남현욱;박종현
    • 동의생리병리학회지
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    • 제23권3호
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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.

Compound 48/80로 유발된 Allergy에 대한 생혈윤부음(生血潤膚飮)의 효과 (Inhibitory Effect of Saingheylyunbooem on Compound 48/80 Stimulated Allergic Reaction)

  • 김희열;박종현
    • 동의생리병리학회지
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    • 제24권1호
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    • pp.48-54
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    • 2010
  • The present study was conducted to investigate the anti-allergic activity of Saingheylyunbooem(SHU)). We investigated the anti-allergic effects of SHU in RBL-2H3 basophilic leukemia cells by compound 48/80, a mast cell degranulator in mice. SHU inhibited ${\beta}$-hexosaminidase and histamine release from compound 48/80 stimulated RBL-2H3 cells. The in vitro anti-inflammatory activities of SHU in LPS-stimulated RAW 264.7 cells were investigated. SHU inhibited NO production effectively dowregulated the expression of iNOS mRNA and iNOS protein expression in LPS-stimulated RAW 264.7 cells. These result provide evidences that SHU may be beneficial in the treatment of allergic inflammtory disease.