• Korean Journal of Acupuncture
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• v.31 no.1
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• pp.14-19
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• 2014

Objectives : In this report, we investigated the effect of ethanol extract of Syzygium aromaticum(L.) Merr. & Perry.(SAE) on the RBL-2H3 cell-mediated allergic response and studied its possible mechanisms of action. Methods : Cytotoxicity on RBL-2H3 cell was evaluated by MTT assay. Anti-allergic activity of SAE was assessed by ${\beta}$-Hexosaminidase and Histamine secretion, ${\beta}$- Hexosaminidase and Histamine secretion were measured by ELISA assay. Evaluate the mechanisms of effect of SAE on the secretion of degranulate mediators, we examined the effect of SAE on the activation of mitogen-activated protein kinases using western blot analysis. Results : SAE had no cytotoxicity on rat basophilic leukemia cell(RBL-2H3). Moreover SAE dose-dependently inhibited RBL-2H3 cell degranulation and histamine release. SAE specifically blocked the IgE-induced p38 mitogen-activated protein kinase activation. Conclusions : Our findings provide evidence that Syzygium aromaticum ethanol extract inhibits mast cell derived allergic reaction, and also demonstrate the involvement of p38 MAPK phosphorylation.

• Advances in Traditional Medicine
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• v.10 no.3
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• pp.200-207
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• 2010

The fruits of Piper cubeba L. are used traditionally to treat respiratory disorders in Indonesia. In order to determine the compounds responsible for this activity, the fruits were extracted with nhexane followed by ethanol to give n-hexane and ethanol extracts. Based on tracheospasmolytic assay on these two extracts, the n-hexane extract was more active to inhibit trachea contraction than that of ethanol extract. Upon bioassay guided isolation of the n-hexane extract, a tracheospasmolytic active compound was isolated and identified as dihydrocubebin [(3,4),(3',4')-bis-methylenedioxy-9,9'dihydroxylignan] $(\underline{1})$. Compound $\underline{1}$ was tested further for its ability to inhibit histamine released from mast cells, using rat basophilic leukemia (RBL-2H3) cell line and rat peritoneal mast cells RPMCs) as models; and $DNP_{24}$-BSA, thapsigargin, ionomycin, compound 48/80 and PMA were used as inducers for histamine released from mast cell. The test result showed that $\underline{1}$ inhibited histamine release from RBL-2H3 cells induced by $DNP_{24}$-BSA, thapsigargin and ionomycin. In addition, $\underline{1}$ suppressed histamine release from RPMC induced by either thapsigargin or ionomycin. However, $\underline{1}$ did not inhibit histamine release from RPMC induced by either compound 48/80 or combination PMA-sub optimum dose of ionomycin. Therefore, it was concluded that the inhibitory effects of $\underline{1}$ on the histamine released from mast cells may involve mechanisms related to intracellular $Ca^{2+}$ signaling events or downstream processes of intracellular $Ca^{2+}$ signaling in mast cells.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.21 no.3
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• pp.94-103
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• 2008

비만세포의 한 종류인 rat basophilic leukemia (RBL-2H3) 세포를 이용하여 교계사물탕-가감의 항알레르기 효과를 확인하고자 하였다. Phorbol 12-myristate 13-acetate (PMA)와 calcium ionophore A23187을 이용하여 RBL-2H3 세포를 자극한 후 세포의 탈과립 정도를 $\beta$-hexosaminidase assay로 확인한 결과, 전 처리한 교계사물탕-가감의 농도 의존적으로 탈과립을 억제하였다. Pro-inflammatory cytokines인 tumor necrosis factor (TNF)-alpha와 interleukin(IL)-4의 분비량을 enzyme-linked immunosorbent assay (ELISA)로 확인한 결과 교계사물탕-가감의 농도 의존적으로 감소하였으며, 이들 cytokines와 염증 반응에 주요한 인자인 cyclooxygenase (COX)-2 의 mRNA 발현 정도 역시 교계사물탕-가감에 의해 감소함을 확인할 수 있었다. 이러한 실험 결과로 보아 교계사물탕-가감은 알레르기 관련 질환의 치료에 응용될 수 있을 것으로 사료된다.

• The Journal of Pediatrics of Korean Medicine
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• v.32 no.4
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• pp.71-86
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• 2018

Objectives The purpose of this study is to investigate the effects of Gardenia jasminoides for. grandiflora extracts' (GAJ) anti-inflammatory effect on RBL-2H3 mast cells and OVA/alum-sensitized mice. Methods In this study, IL-4 and IL-13 production was measured via ELISA analysis, and mRNA expressions of GM-CSF, IL-4, IL-5, $TNF-{\alpha}$, IL-6 were analyzed by real-time PCR. In addition, MAPKs and $NF-{\kappa}B$ p65 transcription factors were examined using western blotting, and ELISA was used to understand IgE, IL-4, and IL-13 production in ovalbumin-allergic mice in in vitro study. Results As a result of this study, 1. GAJ were observed to suppress the mRNA expression of GM-CSF, IL-4, IL-13, IL-5, $TNF-{\alpha}$, IL-6 in comparison to PMA 50 ng/ml, ionomycin $0.5{\mu}M$ (PI) control group. 2. GAJ also inhibited the IL-4, IL-13 production in comparison to PI control group. 3. Western blot analysis showed decrease on the expression of mast-cell-specific transcription factors, including MAKPs (ERK, JNK, p38) and $NF-{\kappa}B$ p65. 4. Orally-administered GAJ group in OVA/alum induced Balb/c mice showed decreased level of OVA-specific IgE in the serum. This group also has shown decreased the level of IL-4, IL-13 in the splenocyte culture supernatant. Conclusions Obtained results suggest that GAJ may regulate the allergic inflammation by transcription factors MAKPs (ERK, JNK, p38) and $NF-{\kappa}B$ p65 causing inhibition of Th2 cytokines in mast cells and OVA/alum-sensitized mice.

• The Korean Journal of Food And Nutrition
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• v.15 no.4
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• pp.382-385
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• 2002

It is well known that the Petasites japonicum have been used for a long time in traditional medicine for the treatment of allergic diseases such as lacquer poisoning and asthma. Anti-allergic actions of Petasites japonicum extracts were asessed by testing their effects on the degranulation of mast cells. For this, hexosaminidase released (degranulation marker) from RBL-2H3 cells(mast cell line) was used. At the concentration of 300 $\mu\textrm{g}$/mL of the methanol, ethylacetate and hot water extract, the degranulation of RBL-2H3 cells were inhibited 83.33, 69.75 and 35.4%, respectively. These results suggest that the Petasites japonicum could be provide a effective resource for the control of allergic diseases.

• The Journal of Internal Korean Medicine
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• v.43 no.1
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• pp.68-78
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• 2022

Objective: In this study, we investigated the protective effect of rhododendron mucronulatum extract (RME) on allergic reactions and inflammation. Methods: The effect of RME was determined using ELISA and RT-PCR in RBL-2H3 mast cells and RAW 264.7 macrophage cells. We determined cell viability, β-hexosaminidase release, and the synthesis of IL-4 and TNF-α in RBL-2H3 cells. In addition, we determined NO from RAW 264.7 and the gene expression of IL-1β, iNOS, IL-6, TNF-α, and IL-10. Results: RME inhibited β-hexosaminidase release and synthesis of IL-4 and TNF-α in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. In addition, RME inhibited the production of NO and the gene expression of IL-1β, iNOS, IL-6, and TNF-α in LPS-stimulated RAW 264.7 cells. Conclusion: From these results, we concluded that RME possesses anti-allergic activity and anti-inflammatory activity due to the inhibition of mast cells and macrophage function.

• Journal of Mushroom
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• v.12 no.4
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• pp.324-329
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• 2014

For evaluating the anti-allergy activity of mushrooms, forty two mushroom extracts were studied their inhibitory activities on the IgE-mediated degranulation in rat basophilic leukemia cell line (RBL-2H3 cell). Effects of mushroom extracts on the release of interleukin-4 (IL-4) and beta-hexosaminidase, and the cell viability of the IgE-sensitized were measured. From the analysis, five mushroom extracts such as the water extract from Flammulina velutipes (Curt.:Fr.) Sing showed the suppressive activities on IL-4 release as 20%. Eight extracts including the water extract of Ganoserma lucidum showed the suppressive activities on ${\beta}$-hexosaminidase release as 20%. Almost all of the extracts stimulated the proliferation of RBL-2H3 cell. The water extracts of Flammulina velutipes and Phellinus linteus were examined against the inhibitory activity in the production of IL-4 and ${\beta}$-hexosaminidase. Additionally, the extracts from Ganoserma lucidum, Isaria japonica, Phellinus linteus and Pleurotus ostreatus inhibited dose-dependently on ${\beta}$-hexosaminidase production. In conclusion, the result suggests that the mushrooms with the potent inhibitory efficacies on the degranulation of the mast cell would be candidate resources for the anti-allergy resources, and thus need to study for their utilization.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.492-498
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• 2005

The mechanism of activation of phospholipase D2 (PLD2) remains undefined although mechanisms have been described for the activation of PLDI. By expression of mutated forms of haemaglutinnin-tagged PLD2 in a mast cell (RBL-2H3) line, we show that PLD2 is phosphorylated at tyrosines -11, -14, and -470 and that tyrosine-470 is critical for activation of PLD2 by antigen. Studies were performed with mutated-DNA constructs for haemaglutinnin-tagged PLD2 in which codons for tyrosine -11, -14, -165, and -470 were mutated to phenylalanine either individually or collectively. Transient expression of these constructs showed that mutation of tyrosine -11, -14, -470, or all tyrosines (all-mutated PLD2) suppressed antigen-induced tyrosine phosphorylation of PLD2 but only the tyrosine-470 mutant failed to be activated by antigen as assessed by in vitro assay of immunoprepitated PLD2 or by assay of PLD in intact cells. The critical role of tyrosine-470 was confirmed in studies with add-back mutants (phenylalanine back to tyrosine) of the all-mutated PLD. The findings provide the first description of a mechanism of activation of PLD2 in a physiological setting.

• Journal of Applied Biological Chemistry
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• v.53 no.3
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• pp.139-146
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• 2010

The inhibitory effects of 25 polyphenols against in vitro allergic reactions were compared using biochemical and cell assays. Three polyphenols including curcumin, gallic acid, and quercetin suppressed the release of $\beta$-hexosaminidase from ionophore A23187-stimulated RBL-2H3 cells more effectively (>50% inhibition at $100{\mu}M$ concentration). They were found to have potencies in suppressing the release of histamine not only from ionophore A23187-, but also from immunoglobulin E (IgE)-stimulated RBL-2H3 cells. Moreover, such suppressive effects of the three polyphenols were also observed in A23187 plus PMA-costimulated rat peritoneal mast cells. The extent of inhibition were quantified as the respective polyphenol concentration that inhibit 50% ($IC_{50}$) of $\beta$-hexosaminidase or histamine release, showing an inhibition tendency with decreasing order of curcumin>gallic acid>quercetin. Down-regulation of $Ca^{2+}$ influx was suggested as the cause of the inhibition of $\beta$-hexosaminidase and histamine releases in these cells. The immune process inhibition was confirmed by the observed reduction in the gene expressions and release of pro-inflammatory cytokine tumor necrosis factor (TNF)-$\alpha$, interleukin (IL)-$1\beta$, and IL-4, due probably to antioxidant activity of the polyphenols. These findings illustrate that curcumin, gallic acid, and quercetin may be beneficial against allergic inflammatory diseases.

• Journal of Ginseng Research
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• v.43 no.2
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• pp.282-290
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• 2019

Background: Ginsenoside Rg3 (G-Rg3) is the major bioactive ingredient of Panax ginseng and has many pharmacological effects, including antiadipogenic, antiviral, and anticancer effects. However, the effect of G-Rg3 on mast cell-mediated allergic inflammation has not been investigated. Method: The antiallergic effects of G-Rg3 on allergic inflammation were evaluated using the human and rat mast cell lines HMC-1 and RBL-2H3. Antiallergic effects of G-Rg3 were detected by measuring cyclic adenosine monophosphate (cAMP), detecting calcium influx, and using real-time reverse transcription polymerase chain reaction, enzyme-linked immunosorbent assay, Western blotting, and in vivo experiments. Results: G-Rg3 decreased histamine release from activated mast cells by enhancing cAMP levels and calcium influx. Proinflammatory cytokine production was suppressed by G-Rg3 treatment via regulation of the mitogen-activated protein kinases/nuclear factor-kappa B and receptor-interacting protein kinase 2 (RIP2)/caspase-1 signaling pathway in mast cells. Moreover, G-Rg3 protected mice against the IgE-mediated passive cutaneous anaphylaxis reaction and compound 48/80-induced anaphylactic shock. Conclusion: G-Rg3 may serve as an alternative therapeutic agent for improving allergic inflammatory disorders.