• Biomolecules & Therapeutics
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• 제9권4호
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• pp.282-284
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• 2001

The antiallergic drug, cetirizine, inhibits the histamine release from a rat basophilic leukemia (RBL-2H3) cell line, which is frequently used as a mast cell model. By investigating inhibitory activities of (+)- and (-)-cetirizine in RBL-2H3 cells on the histamine release, we aimed to evaluate the effect of their structual characteristics on the antihistamine activity. The study on RBL-2H3 cell has clearly demonstrated that the (-)-cetirizine is significantly more potent than the (+)- or the racemic cetirizine, although there was no difference in pharmacokinetics between (+)- and (-)-cetirizine in rats.

• 대한본초학회지
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• 제34권1호
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• pp.23-31
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• 2019

Objectives : The aim of this study was to investigate the effect for allergic-inflammation of Fritillariae Thunbergii Bulbus (FTB) on HaCaT cells and RBL2H3 cells. Methods : To investigate the effects of FTB for anti-inflammation in HaCaT cells, the cells were pretreated with FTB for 1h and then stimulated with $TNF-{\alpha}/IFN-{\beta}$ for 24h. Then thymus and activation-regulated chemokine (TARC) and Macrophage-derived chemokine (MDC) levels were analyzed with ELISA kit. Also to investigate the effect of skin barrier protein, the cells were treated with FTB of various concentrations, and then cells were harvested, expressions of skin barrier protein were measured with RT-PCR. To investigate the effects of FTB for anti-allergy in RBL2H3 cells, the cells were pre-treated with FTB for 1h, and then stimulated with A23187 for 30 min. ${\beta}$-hexosaminidase, IL-4 and $TNF-{\alpha}$ were measured using cultured media. The cells were harvested to analyze the mechanism of the effect for FTB via Western blot. Results : FTB did not show cytotoxicity in HaCaT and RBL2H3. In HaCaT cells, FTB significantly suppressed the expression of TARC, MDC at a dose-dependent manner and markedly increased formation of the skin barrier proteins. In RBL2H3 cells, FTB decreased release of the ${\beta}$-hexosaminidase, IL-4 and $TNF-{\alpha}$ in RBL2H3 through inhibition of the phosphorylation of JNK and p38, which are include in the signaling mechanism of MAPK Conclusion : These results indicate that FTB has an anti-inflammatory effect on the allergic response through blocking MAPK pathway. This suggest that FTB could be a therapeutic agent for allergic response.

• 대한한의학방제학회지
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• 제29권4호
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• pp.191-203
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• 2021

Objectives : In this study, we investigate the anti-allergic effects of Ullmus macrocarpa Hance (Ulmus) on RBL-2H3 mast cell (basophilic leukemia cell line), which are mediated by FcεRIs. Methods : We evaluated the effect of the ethanol extract of Ulmus on the allergic inflammatory response in IgE-antigen-mediated RBL-2H3 cells. Cell toxicity was determined by MTT assay and the markers of degranulation such as beta-hexosaminidase, histamine, PGD2, TNF-α, IL-4, IL-6 production of inflammatory mediators and FcεRI-mediated protein expression by western blot. Results : Ulmus inhibited degranulation and production of allergic mediators (e.g., TNF-α, IL-4, and IL-6) in them. Ulmus reduced histamine levels, expression of FcεRI signaling-related genes such as Lyn, Syk, and Fyn, and extracellular signal-regulated kinase phosphorylation in mast cells. Also, Ulmus reduced PGD2 release and cyclooxygenase-2 expression, and cytosolic phospholipase A2 phosphorylation in FcεRI-mediated RBL-2H3 mast cells. Conclusions : These results indicate that Ulmus exhibits anti-allergic activity through inhibition of degranulation and inflammatory mediators and cytokine release. These findings suggest that Ulmus may have potential as a prophylactic and therapeutic agent for the treatment of various allergic diseases.

• 한방안이비인후피부과학회지
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• 제21권3호
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• pp.82-93
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• 2008

Objective: Previously, the methanol extracts of the semen of Xanthium strumsrium could involved anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated Raw 264,7 cells, We evaluated the anti-allergic effects of X. strumarium on rat basophilic leukemia (RBL-2H3) cells, Methodes : To investigate the effect of X. strumarium on the phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187-induced RBL-2H3 cells. The effects of X. strumarium on the degranulation and the pro-inflammatory cytokines secretion and expression from RBL-2H3 cells were evaluated with $\beta$-hexosaminidase assay, ELISA, and RT-PCR analysis, In addition, we examined the effects of X. strumarium on nuclear factor (NF)-${\kappa}B$ activation and $I{\kappa}B-\alpha$ degradation using Western blot analysis. Results : X. strumarium inhibited degranulation and secretions and expressions of pro-inflammatory cytokines, such as tumor necrosis factor-alpha ($TNF-\alpha$), interleukin (IL)-4 and cyclooxygenase (COX)-2, on stimulated RBL-2H3 cells, however, X. strumarium not affect cell viability. In stimulated RBL-2H3 cells, the protein expression level of nuclear factor-kappa B (NF-${\kappa}B$) was decreased in the nucleus by X. strumarium. In addition, X. strumarium suppressed the degradation of inhibitory protein $I{\kappa}B-{\alpha}$ protein in RBL-2H3 cells. Conclusion : These results suggest that X. strumarium inhibits the degranulation and secretion of pro-inflammatory cytokines through blockade of NF-${\kappa}B$ activation and I $I{\kappa}B-{\alpha}$ degradation.

• 동의생리병리학회지
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• 제26권6호
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• pp.915-921
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• 2012

Artemisiae Asiaticae Herba (AAH) has been used to remedy of symptoms such as bleeding, dysmenorrhea, eczema and itchy skin in Oriental Medicine. In this study, we investigated the protective effect of AAH on allergic response. The effect of AAH was analyzed by ELISA and RT-PCR in RBL-2H3 cells. We investigated cell viability, ${\beta}$-hexosaminidase and histamine as markers of degranulation, production of IL-4 and TNF-${\alpha}$, and gene expression of HDC2, cytokines and $Fc{\varepsilon}RI$ ${\alpha}{\beta}{\gamma}$ subunit. We found that AAH suppressed ${\beta}$-hexosaminidase and histamine release, the production of IL-4 and TNF-${\alpha}$ in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. AAH also significantly decreased cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-12, IL-13, TNF-${\alpha}$, and GM-CSF, and increased cytokine mRNA expressions of IL-10 in RBL-2H3. In addition, AAH suppressed mRNA expression of $Fc{\varepsilon}RI$ ${\alpha}{\beta}{\gamma}$ subunit on cell surface. Our results indicate that AAH protects against allergic response and exerts an anti-inflammatory effect through the inhibition of degranulation and production of cytokines and expression of $Fc{\varepsilon}RI$ ${\alpha}{\beta}{\gamma}$ subunit.

• 대한한의학방제학회지
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• 제31권4호
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• pp.315-325
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• 2023

Objectives : Recently, research has been actively conducted on the efficacy of complexes based on oriental medicine prescriptions for improving immune activity and allergies. In this study, In this study, we aimed to examine the effect of Angelica gigas Nakai and Corni fructus extract (AC), medicinal herbs, among candidate drugs derived through preliminary experiments with various components of oriental medicine prescriptions for allergies, on allergies in RBL-2H3 cells. Methods : We evaluated the effect of the ethanol extract of Ulmus on the allergic inflammatory response in anti-DNP-IgE activated DNP-HSA in RBL-2H3 cells. Cell toxicity was determined by WST-1 assay and the markers of degranulation such as beta-hexosaminidase, histamine, TNF-α and IL-6 production of inflammatory mediators and FcεRI-mediated expression. Results : The results showed that treatment with AC extract (20, 40 and 80㎍/㎖) noncytotoxic levels and significantly inhibited the release of β-hexosaminidase, histamine and the production of TNF-α and IL-6 in RBL-2H3 by the antigen stimulation. Conclusions : These results indicate that AC extract exhibits anti-allergic activity through inhibition of degranulation and inhibition of inflammatory mediators and cytokine release. These findings suggest that AC extract may have potential as a prophylactic and therapeutic agent for the treatment of various allergic diseases.

• Journal of Applied Biological Chemistry
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• 제64권1호
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• pp.19-23
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• 2021

전통생약으로서 다양한 효능과 향신료, 조미료로서 사용되어온 초과(草果)의 항알레르기 성분연구를 위해 80% 에탄올 추출물 중 CH2Cl2, 분획으로부터 3 종의 flavonoid를 단리하였으며, 이들의 물리화학적 성상과 분광분석 데이터로부터 구조를 확인한 결과 naringenin-5-O-methyl ether (1), helichrysetin (2) 및 cardamomin (3)로 각각 동정하였다. 이들 화합물 중 화합물 2는 Amomum 속으로부터 처음 규명되었고, 화합물 3은 이 식물에서 처음 보고되는 화합물이다. 또한 화합물 2와 3은 RBL-2H3 세포로부터 β-hexosaminidase assay를 수행한 결과 탈과립 억제효과를 나타내었다. 따라서 초과의 chalcone 성분은 탈과립 억제효능을 통하여 알러지 질환에 유용할 것으로 사료된다.

• 한방안이비인후피부과학회지
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• 제25권3호
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• pp.1-12
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• 2012

Objectives : Fagopyrum esculentum(FE) has been used for removal of inflammation of internal organs and treatment of sore and ulcer by heat toxin in Korean herbal medicines. In this study, To investigated the protective effect of FE on allergic response, we determined whether FE inhibits allergic response. Methods : The effect of FE was analyzed by ELISA, RT-PCR and Western blot in RBL-2H3 cells. We investigated cell viability, ${\beta}$-hexosaminidase, as a marker of degranulation, cytokne, and intracellular ROS and MAPK and NF-${\kappa}B$ signaling. Results : We found that FE suppressed ${\beta}$-hexosaminidase release, the production of IL-4 and TNF-${\alpha}$ and intracellular ROS level in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. FE also significantly inhibited cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF in RBL-2H3. In addition, PF suppressed the phospholyation of ERK1/2, JNK1/2, p38 and $I{\kappa}B{\alpha}$ and NF-${\kappa}B$ signal transduction pathway. Conclusions : Our results indicate that FE protects against allergic response and exerts an anti-inflammatory effect through the inhibition of degranulation and production of cytokines and ROS via the suppression MAPK and NF-${\kappa}B$ of signal transduction. Abbrevations : FE, Fagopyrum esculentum; RBL-2H3, rat basophilic leukemia cell line; ROS, reactive oxygen species; MAPK, Mitogen-activated protein kinase; $NF{\kappa}B$, nuclear factor ${\kappa}B$; $TNF{\alpha}$, Tumor necrosis factor alpha; GM-CSF, Granulocyte macrophage colony-stimulating factor; ERK, extracellular-signal-regulated kinase; JNK, c-Jun NH2-terminal kinase; p38, p38 MAP kinase; $I{\kappa}B{\alpha}$, inhibitory-kappa B alpha.

• 한국미생물·생명공학회지
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• 제33권3호
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• pp.189-193
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• 2005

RBL-2H3 cell lysates에 anti-protein phosphatase(PP) 1, 2A, 2B 항체를 첨가한 후 immunoprecipitation을 실시한 결과 PP2B를 가해준 샘플에서만 HRF를 확인하였다. 역으로 monoclonal anti-HRF 항체를 가한 후 immunoprecipitation을 실시한 결과 PP1, 2A는 검출되지 않았으나 PP2B의 경우는 regulatory subunit(19 kDa), catalyic subunit(60 kDa) 모두 확인할 수 있었다. Affinity chromatography를 통해서도 PP2B가 HRF의 탈인산화에 관여함을 확인하였다 즉 19kDa의 PP2B regulatory subunit과 60kDa의 catalytic subunit 모두가 확인되었으며 외부 $Ca^{2+}$이온 첨가 여부에 따른 차이는 관찰할 수 없었다. 결론적으로 RBL-2H3 cell에서 PP2B는 PP1이나 PP2A에 비해 상대적으로 그 존재량은 적으나 HRF와 상호작용하는 phosphatase로서 검출된 반면 PP1이나 PP2A는 검출되지 않았다.

• 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.