• Archives of Pharmacal Research
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• v.25 no.5
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• pp.675-680
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• 2002

In order to investigate the underlying mechanism of HCI in oesophagitis, the inflammatory response to HCI was observed in RBL-2H3 mast cells. Rat basophilic leukemia (RBL-2H3) cells were used to measure histamine release, arachidonic acid (AA) release, reactive oxygen species (ROS) and peroxynitrite generation induced by HCI. Exogenous HCl increased the level of histamine release and ROS generation in a dose dependent manner, whereas it decreased the spontaneous release of [$^3$H] M and the spontaneous production of peroxynitrite. Mepacrine (10 $\mu$M), oleyloxyethyl phosphorylcholine (10 $\mu$M) and bromoenol lactone (10 $\mu$M) did not affect both the level of histamine release and ROS generation induced by HCI. U73122 (1 $\mu$M), a specific phospholipase C (PLC) inhibitor did not have any influence on level of histamine release and ROS generation. Propranolol (200 $\mu$M), a phospholipase D (PLD) inhibitor, and neomycin (1 mM), a nonspecific PLC and PLD inhibitor, significantly inhibited both histamine release and ROS generation. Diphenyleneiodonium (10 $\mu$M), a NADPH oxidase inhibitor, and tiron (5 mM), an intracellular ROS scavenger significantly inhibited the HCI-induced histamine release and ROS generation. These findings suggest that the inflammatory responses to HCI is related to histamine release and ROS generation, and that the ROS generation by HCI may be involved in histamine release via the PLD pathway in RBL-2H3 cells.

• Microbiology and Biotechnology Letters
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• v.33 no.3
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• pp.231-235
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• 2005

Histamine-releasing factors (HRFs) are soluble mediators that can release histamine and other mediators from basophils and mast cells and their activity can vary, depending on the type of IgE. The activity of HRFs is affected by the presence of IgE, although HRF is thought to bind to a specific receptor other than IgE. Until now, HRF signaling pathway including its receptor remains unclear in spite of numerous studies. Since there had been many reports about reactive oxygen species (ROS) as a signaling molecule rather than as a by-product of metabolism, we investigated the possibility of ROS as an intracellular messenger involved in HRF-mediated histamine degranulation. In RBL-2H3 cells, ROS was generated by HRF using $H_2O_2$-sensitive fluorescence of fluorescent 2', 7'-dichlorofluorescein ($H_2DCFDA$). These effects were blocked by anti-oxidant N-acetylcysteine (NAC). These results suggest that ROS generation could play a role as an intracellular messenger in histamine release by HRF.

• Natural Product Sciences
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• v.27 no.4
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• pp.251-256
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• 2021

Lobelia chinensis Lour. has commonly been used in traditional Chinese medicine for the treatment of antidote, diuretic, diarrhea, and inflammation. This study aimed to identify the active compounds in an aqueous extract of L. chinensis responsible for its anti-atopic effect in vitro using RBL-2H3 cells. A chemical investigation of secondary metabolites in an aqueous extract of L. chinensis led to the isolation of nine chemical constituents, which included the four marker compounds, and these were evaluated for their inhibitory effects on IL-4 mRNA expression and the release of β-hexosaminidase in propidium iodide-induced RBL-2H3 cells. We found diosmetin and fraxidin inhibited cellular IL-4 mRNA expression, and that diosmetin and 6,8-dimethoxycoumarin inhibited DNP-specific IgE-induced degranulation in these cells. Our study suggests that diosmetin, fraxidin, and 6,8-dimethoxycoumarin are potential candidates for the treatment of atopic diseases.

• The Journal of Pediatrics of Korean Medicine
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• v.24 no.2
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• pp.126-136
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• 2010

Objectives The purpose of this study was to investigate the antiallergic and antioxidative effects of Chaenomeles Sinensis (CS). Methods CS pretreatments inhibited anti-DNP IgE in RBL-2H3 mast cells for an hour. we measured cell viability, $\beta$-hexosaminidase release, IL-4, TNF-$\alpha$ secretion, and IL-4, TNF-$\alpha$ mRNA expression CS pretreatments inhibited DNP-HSA($10\;{\mu}g/ml$) for ten minutes, we measured Dicholrodihydrofluorescein(DCF) and DPPH radical-scavenging activity in 0.5 mM 1,1-diphenyl-2-picrydrazyl(DPPH) radical solution, 0.1ml, 99% ethanol 0.8ml, and CS 0.1 ml mixed solution. Results 0, 1, 2, 3, 4, 5 mg/ml CS treatments were not affect on cell viability and inhibited b-hexosaminidase release, IL-4, TNF-$\alpha$ secretion, CS treatments also decreased IL-4, TNF-$\alpha$ mRNA expression in RBL-2H3 cells. CS treatments inhibited reactive oxygen species(ROS) and DPPH radical-scavenging activity. Conclusions These results suggest that CS may be useful for the prevention or treatment of allergic disease.

• Korean Journal of Oriental Medicine
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• v.16 no.3
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• pp.155-166
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• 2010

Objetives : The purpose of this study was to examine the pathway of anti-allergic effects of Aconiti Ciliare Tuber (ACT). Methods : We examined cell viability, ${\beta}$-hexosaminidase release, pro-inflammatory cytokines secretion and mRNA expressions, nuclear factor-kappa B (NF-${\kappa}B$) (p65) activation, inhibitor kappa B-alpha ($I{\kappa}B-{\alpha}$) degradation, and MAPKs activation from RBL-2H3 cells pre-treatment by ACT of 1.0 mg/ml, 2.0 mg/ml separately. Results : We observed that ACT reduced the secretion of ${\beta}$-hexosaminidase, TNF-${\alpha}$, IL-4 and the expression of COX-2 mRNA in RBL-2H3 cells. Futhermore, ACT inhibited the levels of activation of NF-${\kappa}B$ (p65) protein, ERK MAPK, and degradation of $I{\kappa}B-{\alpha}$ in RBL-2H3 cells. Conclusions : These results show that ACT has an anti-histamine effect and inhibitory effect of NF-${\kappa}B$ (p65) through regulation of $I{\kappa}B-{\alpha}$ degradation. This improves that ACT could be used as an anti-allergic medicine.

• Korean Journal of Food Science and Technology
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• v.41 no.6
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• pp.717-721
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• 2009

The anti-allergic effect of the methanol extract of Zizania latifolia Turcz was investigated. Methanol extract of Z. latifolia Trucz did not display any cytotoxicity activities in the IgE-sensitized RBL-2H3 cells. The extract was found to inhibit compound 48/80-induced degranulation and antigen-induced $\beta$-hexosaminidase release as well as PMA plus A23187-induced TNF-$\alpha$ production in RBL-2H3 mast cells in a dose-dependent manner. Release of $\beta$-hexosaminidase, a marker for the release of histamine in mast cells, was inhibited with an $IC_{50}$ value of $73.8\;{\mu}g/mL$. These results suggest that Z. latifolia extracts may be useful for the prevention of type I allergic reaction.

• YAKHAK HOEJI
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• v.58 no.5
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• pp.294-299
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• 2014

In this study, we investigate anti-allergic and anti-inflammatory effects of Leonurus sibiricus seed (LSS) extract in basophilic leukemia RBL-2H3 cells. To identify anti-allergic actions of LSS, the degranulation was evaluated in IgE and DNP-BSA stimulated RBL-2H3 cells. At the concentration of $100{\mu}g/ml$ of methanol (MeOH) extract and Methylene chloride (MC) and Ethyl acetate (EtOAc) fractions, the degranulation was significantly inhibited 16.7%, 16.7% and 27.9% respectively. And then, to assess anti-inflammatory effects of LSS, IL-4 and IL-13 mRNA level were detected in PMA/ionomycin (PI)-induced RBL-2H3 cells and cell proliferation and IL-4 mRNA level in isolated splenocytes from Balb/c mice. LSS MeOH extract and MC and EtOAc fractions significantly decreased the level of IL-4 and IL-13 mRNA in PI-induced RBL-2H3 cells and showed inhibitory effects on cell proliferation and expression of IL-4 mRNA level in mouse splenocytes. Taken together, these results suggest that LSS has potential anti-allergic and anti-inflammatory effects and EtOAc fraction is the most effective in regulating immune responses.

• Korean Journal of Plant Resources
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• v.25 no.3
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• pp.339-345
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• 2012

The marine carotenoid fucoxanthin can be found in marine brown seaweeds, macroalgae, diatoms, and microalgae, and has remarkable biological properties. Numerous studies have shown that fucoxanthin has considerable potential and promising applications in human health, but the underlying mechanisms involved in its anti-allergic activity are not fully understood. We here investigated the mechanisms by anti-allergic activity of fucoxanthin fraction from Eisenia bicyclis in immunoglobulin E-antigen complex (IgE/DNP-BSA)-stimulated RBL-2H3 mast cells. This study we found that the fucoxanthin inhibits the release of ${\beta}$-hexosaminidase and suppressed not only transcriptional activation of NF-${\kappa}B$, but also phosphorylation of ERK and JNK in IgE/DNP-BSA-treated RBL-2H3 cells. Fucoxanthin may be useful for preventing allergic diseases, including asthma and atopic dermatitis.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.25 no.3
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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.

• The Journal of Pediatrics of Korean Medicine
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• v.25 no.1
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• pp.28-39
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• 2011

Objectives: SaengRyoSaMulTang is a herbal formula in Oriental Medicine, known anti-allergens. However, its mechanism and cellular targets have not been found yet. Thus the study has developed to investigate the suppressive effect of SaengRyoSaMulTang. Methods: In the study, cellular viability, IL-4, IL-13 mRNA expression, IL-4, IL-13 production, manifestations of GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-${\kappa}$B p65 transcription factors were examined by Real-Time PCR, ELISA analysis and western blotting. Results: As a result of treating with SaengRyoSaMulTang extract(SRSMT), the study has shown that the amount of Th2 cytokines, which include PI induced IL-4 and IL-13, plays a significant role in suppressing effect. RBL-2H3 mast cells significantly suppressed the PI-induced Th2 cytokine production including IL-4 and IL-13 in a dose dependent manner. PI-induced IL-4 and IL-13 production was significantly suppressed by SRSMT intervention. Western blot analysis of transcription factors involving IL-4 and IL-13 expression also revealed a prominent decreases of mast cell's specific transcription factors including GATA-1, GATA-2, NF-AT2, c-Jun and c-Fos, but NF-${\kappa}$B p65. Conclusions: The study suggests that the anti-allergenic activities of SRSMT may regulate the transcription factors GATA-1, GATA-2, NF-AT2, c-Jun and c-Fos inhibiting Th2 cytokines IL-4 and IL-13 in mast cells.