• 한국식품영양학회지
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• 제15권4호
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• pp.382-385
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• 2002

본 연구에서는 민간에서 옻나무에 의 한 알레르기반응 및 기관지천식의 치료 효과가 있는 것으로 알려진 머위(Petasites japonicum)추출물의 항알레르기 효과를 RBL-2H3 세포로부터 hexosaminidase의 방풀 억제 정도를 측정하는 enzyme assay를 이용하여 검색하였다. 실험결과에 대한 오차범위가 낮은 추추출 300 $\mu\textrm{g}$/mL 농도의 수준에서 methanol 추출물은 83.33%, ethylacetate 추출물은 69.75%, 열수 추출물은 50.62%의 저해효과를 나타냈으며, 대조구로 사용한 quercetin은 같은 농도에서 85.80%의 저해 효과를 나타냈다. 따라서 정제된 quercetin의 hexosaminidase의 방출 억제 활성을 비교해 볼 때 머위 추출물이 조추출물 상태인 점을 고려한다면 상당한 항알레르기 효과가 있음을 시사하고 있어 새로운 항알레르기성 신기능 물질로의 이용방안을 탐색하는 기초자료가 되리라 사교된다.

• 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.

• 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.

• 대한통합의학회지
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• 제8권4호
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• pp.163-169
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• 2020

Purpose : This study aimed to investigate the anti-allergic effect of Persicaria perfoliata water extract (PPWE) on IgE stimulated rat basophilic leukemia (RBL-2H3) cell line. Methods : P. perfoliata (L.) H. Gross has been used in traditional medicine as an anti-allergic agent, antipyretic, and diuretic and for respiratory disorders. To analyze the anti-allergic activity of PPWE, release of β-hexosaminidase in RBL-2H3 cells was estimated by enzyme linked immunosorbant assay (ELISA). Also, the cytotoxic effect of PPWE was identified by WST assay, and nuclear factor (NF)-κB and its upstream signaling molecules were assessed by western blot analysis. Results : PPWE treatment significantly attenuated β-hexosaminidase release in a dose dependent manner without any cytotoxicity. PPWE inhibited β-hexosaminidase activity by 38.4±1.2, 36.6±0.6, 32.5±2.2 and 26.5±1.2 at 500, 250, 100, and 50 ㎍/㎖ of PPWE, respectively, compared with the control group. In addition, an analysis of the expression level of NF-κB, an inflammation transcription factor, in RBL-2H3 cells upon IgE stimulation provided reults consistent with the results of β-hexosaminidase release. The phosphorylated status of upstream signaling molecules for transcription factor, mitogen activated protein kinases (MAPKs), was also analyzed. The results showed that PPWE treatment dose-dependently inhibited phosphorylation of extracellular regulatory kinase (ERK) and c-Jun N-terminal kinase (JNK). These results show that PPWE had a strong IgE-mediated degranulation inhibitory effect on RBL-2H3 cells. Conclusion : P. perfoliata ameliorated IgE-mediated allergic reaction via the modulation of MAPK and NF-κB signaling pathway in RBL-2H3 cells. These results indicate that P. perfoliata could be a potential candidate for a treatment strategy against various allergic disorders.

• 한국약용작물학회지
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• 제18권4호
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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.

• 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.

• 한국식품과학회지
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• 제30권2호
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• pp.468-471
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• 1998

밤나무 잎을 이용하여 차(茶)로 제조한 다음, 제조된 차에서 항알레르기 효과를 실험하였다. 밤나무 잎 차는 대표적인 차의 형태인 불발효차와 반발효차의 제조공정을 이용하여 증제차와 유념차로 제조하였으며, 항알레르기 효과는 제조된 증제차와 유념차의 각각의 물추출물을 RBL-2H3 세포로부터 hexosaminidase의 방출 억제 정도를 측정하는 enzyme assay를 이용하여 실시한 결과, 물추출물의 $300\;{\mu}/mL$의 수준에서 증제차는 50.4%의 저해효과를 나타냈으며, 유념차는 35.4%의 저해효과를 나타내어 항알레르기 효과가 있음을 확인하였다.

• 한국자원식물학회지
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• 제28권3호
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• pp.321-328
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• 2015

Alpiniae officinarum Rhizoma (the rhizome of Alpinia officinarum Hance, known as lesser galangal), a family of Zingiberaceae, has been used to reduce pain of infection and inflammatory diseases in Asian countries. The present study was focused to evaluate the inhibitory degranulation effect of Alpiniae officinarum Rhizoma extract in RBL-2H3 rat basophilic leukemia cells. Cell viability was measured by MTT assay. RBL-2H3 cells were stimulated by phorbol 12-myristate 13-acetate and calcium ionophore A23187. Mast cell degranulation was analyzed by measuring release of β-hexosaminidase in RBL-2H3 cell. Gene expression was measured by qRT-PCR and signaling molecules were detected by immunoblotting. The Alpiniae officinarum Rhizoma extract suppressed β-hexosaminidase release in dose-dependent manner and inhibited cycloxygenase-2 and tumor necrosis factor-α gene expression. Furthermore, it was found that Alpiniae officinarum Rhizoma extract reduced mitogen-activated protein kinases, especially phosphorylated p38, at 0.75 ㎎/㎖ of Alpiniae officinarum Rhizoma extract concentrations. These data show that Alpiniae officinarum Rhizoma extract has immunosuppressive effect in mast cell induced allergic inflammation.

• 대한본초학회지
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• 제26권4호
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• pp.1-7
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• 2011

Objectives : In this study, we investigated the effect of modified-Okbyungpoongsan (mOP) on mast cell-mediated allergic response in basophilic leukemia cell line, RBL-2H3 mast cells. Methods : Cells were stimulated with anti-DNP-IgE after the treatment of DNP-HSA (AI/D), and then incubated with different concentrations of mOP (0.1, 0.2, 0.5 and 1 mg/$m{\ell}$) in RBL-2H3 cells. Cell toxicity was determined by WST-1 assay. The degranulation of mast cells was observed by microscope with toluidine blue staining and also the levels of beta-hexosaminidase, histamine and TNF-alpha were measured in culture supernatants by enzyme-linked immunosorbant assay. Results : mOP inhibited anti-DNP-IgE-imduced degranulation of mast cells in RBL-2H3 cells. mOP also significantly decreased the levels of histamine and inflammatory cytokine, TNF-alpha in RBL-2H3 cells, but slightly decreased the level of beta-hexosaminidase. Conclusions : These results indicate that mOP, an oriental prescription could be inhibit the allergic response through suppressing the mast cell activation.

• 한방안이비인후피부과학회지
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• 제19권3호통권31호
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• pp.13-21
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• 2006

Object : We investigated the effect of Gamiokyaek-tang (GOYT) on immediate type hypersensitivity. Methods : We investigated anti-dinitrophenyl (DNP) IgE-mediated passive cutaneous anaphylaxis and acetic acid-induced vascular permeability in rodents. Also perfonned MTT assay and b-hexosaminidase activity is measured in RBL-2H3. Results : GOYT inhibited passive cutaneous anaphylaxis and acetic acid-induced vascular permeability by oral administration. All the concentrations of GOYT from 0.1 to 5 mg/ml didn't have an effect on cell viability and cytotoxicity. b-hexosaminidase release in RBL-2H3 was significantly reduced by 2 and 5 mg/ml of GOYT. Conclusion : These results indicate that GOYT have inhibition effects on immediate type hypersensitivity .