• The Korean Journal of Physiology and Pharmacology
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• v.15 no.5
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• pp.313-317
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• 2011

The effects of extremely low frequency electromagnetic fields (EMF) on intracellular $Ca^{2+}$ mobilization and cellular function in RBL 2H3 cells were investigated. Exposure to EMF (60 Hz, 0.1 or 1 mT) for 4 or 16 h did not produce any cytotoxic effects in RBL 2H3 cells. Melittin, ionomycin and thapsigargin each dose-dependently increased the intracellular $Ca^{2+}$ concentration. The increase of intracellular $Ca^{2+}$ induced by these three agents was not affected by exposure to EMF (60 Hz, 1 mT) for 4 or 16 h in RBL 2H3 cells. To investigate the effect of EMF on exocytosis, we measured beta-hexosaminidase release in RBL 2H3 cells. Basal release of beta-hexosaminidase was $12.3{\pm}2.3%$ in RBL 2H3 cells. Exposure to EMF (60 Hz, 0.1 or 1 mT) for 4 or 16 h did not affect the basal or $1{\mu}m$ melittin-induced beta-hexosaminidase release in RBL 2H3 cells. This study suggests that exposure to EMF (60 Hz, 0.1 or 1 mT), which is the limit of occupational exposure, has no influence on intracellular $Ca^{2+}$ mobilization and cellular function in RBL 2H3 cells.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.21 no.3
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• pp.69-81
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• 2008

Objective : The purpose of this study was find out the anti-allergic effects of SESHINGO on the allergic rhinitis. Methods : Cell viability was evaluated by the MTT assay. Cells were treated with the indicated concentration of SSG. $TNF-\alpha$ concentration was measured from cell supernatants using ELISA method. IL-4 concentration was measured from cell supernatants using ELISA method.$IFN-\gamma$ concentration was measured from cell supernatants using ELISA method. Total RNA was isolated, $TNF-\alpha$ and IL-4 mRNA expression was detected by RT-PCR analysis Total RNA was isolated, COX-1, COX-2 mRNA was analyzed by RT-PCR analysis. Results : 1. $\beta$-hexosaminidase release in RBL-2H3 cells were decresed by SESHINGO attentively. 2. Secreting ration of $TNF-\alpha$was restrained in RBL-2H3 cells by SESHINGO attentively. 3. Secreting ration of IL-4 was restrained in RBL-2H3 cells by SESHINGO attentively 4. Revelation of $TNF-\alpha$ mRNA was decresed in RBL-2H3 cells as concetration. 5. Revelation of.IL-4 mRNA was decresed in RBL-2H3 cells as concetration. 6. Revelation of COX-2 mRNA was decresed in RBL-2H3 cells. Conclusion : According to above results, to evaluate the effect of SSG on the anti-allergic effects.

• The Korea Journal of Herbology
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• v.26 no.4
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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.

• The Korea Journal of Herbology
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• v.32 no.6
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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.

• Journal of The Korean Society of Integrative Medicine
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• v.8 no.4
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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.

• The Korea Journal of Herbology
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• v.34 no.1
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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.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.21 no.3
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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.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.12
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• pp.1662-1667
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• 2011

Anthocyanins belong to a group of flavonoid compounds and are well known for their various health beneficial effects, which include antioxidative activities. Among them, the major anthocyanins isolated from seed coat of black soybean (Glycine max L.) were previously characterized as glycosides containing glucopyranose. Asthma is an allergic disease that is strongly associated with various immune cells, including basophils and mast cells. Eosinophils, basophils, and mast cells play important roles in allergic asthma through the release of inflammatory mediators such as asthma-specific T-helper 2 (Th2) cytokines and subsequent amplification of asthma symptoms via degranulation. Rat basophilic leukemia RBL-2H3 cells are the most common in vitro models for evaluating allergic reactions. In this study, we examined the effects of anthocyanin from seed coat of black soybean on antigen-stimulated degranulation and Th2 cytokine production in RBL-2H3 cells. Cell degranulation was evaluated by measuring the release of ${\beta}$-hexosaminidase. ${\beta}$-Hexosaminidase release and Th2 cytokine production in RBL-2H3 cells was much higher upon stimulation with IgE-antigen complex than those in untreated control cells. Anthocyanins significantly suppressed IgE-antigen complex-induced degranulation of RBL-2H3 cells and inhibited IgE-antigen complex-mediated interleukin (IL)-4, IL-13, and tumor necrosis factor ${\alpha}$ (TNF-${\alpha}$) production in RBL-2H3 cells. These findings suggest that anthocyanins from seed coat of black soybean effectively inhibit allergic reactions and may have beneficial effects against allergic asthma.

• YAKHAK HOEJI
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• v.57 no.5
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• pp.309-315
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• 2013

Cyanidin-3-glucoside (C3G), an anthocyanin, is one of the most widespread dietary flavonoids. We investigated the effects of C3G in PMA/ionomycin (PI)-induced RBL-2H3 cells. C3G inhibited the production of IL-4 and IL-13 and also decreased the level of mRNA in a dose-dependent manner. Furthermore, western blot analysis implied that C3G down-regulated the protein level of c-Jun, NF-ATc1 and NF-${\kappa}B$ but not c-Fos. Taken together, we suggest that C3G may have suppressive effects on Th2 cytokines and will be studied further to develop as functional foods that help alleviate allergy symptoms.

• Journal of Applied Biological Chemistry
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• v.58 no.3
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• pp.287-290
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• 2015

Soyasaponins are glycosylated, which gives rise to a wide diversity of structures and functions. We evaluated for inhibitory effects of 4 soyasaponins on antigen-induced degranulation in RBL-2H3 Cells. 4 soyasaponins had shown dose-dependently inhibited histamine and ${\beta}$-hexosaminidase.