• 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.

• 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.

• 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.

• Korean Journal of Acupuncture
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• v.26 no.1
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• pp.125-137
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• 2009

Objectives : We studied anti-allergic effects of Cinnamomi Ramulus(CR) herbal acupuncture and Cinnamomi Ramulus extract. Methods : In vivo, animals were herbal-acupunctured with CR at both ST36s three times for 5 days. Then, we induced active systemic anaphylatic shock using compound 48/80 in ICR mice, and passive cutaneous anaphylaxis using anti-DNP IgE in Sprague Dawley rat. In vitro, we measured cell viability, ${\beta}$ -hexosaminidase release and the expressions of IL-4, TNF-${\alpha}$ and COX-2 mRNA in RBL-2H3 cells after treatment of various concentrations of CR extract. Results : In vivo, CR herbal acupuncture pretreatments at both ST36s inhibited compound 48/80-induced active systemic anaphylatic shock. Passive cutaneous anaphylaxis was inhibited by CR herbal acupuncture pretreatments at both ST36s and optional points. In vitro, CR extract treatments did not affect on cell viability and inhibited ${\beta}$-hexosaminidase release. CR extract treatments also decreased the expressions of IL-4, TNF-${\alpha}$ and COX-2 mRNA in RBL-2H3 cells. Conclusions : These results suggest that CR herbal acupuncture and CR extract should be beneficial in the inhibition of allergic inflammatory response.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.3
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• pp.600-606
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• 2008

The purpose of this study was to investigate Anti-allergic Inflammatory Effects of Hwangyounhaedog-tang on PMA plus A23187-stimulated RBL-2H3 cells. We examined MTT assay, ${\beta}$-hexosaminidase activity, IL-4 and TNF-${\alpha}$ from RBL-2H3 after pre-treatment with HHT. In addition, we investigated expression of COX-1 and COX-2 mRNA using RT-PCR study. The results are as follows: All the concentrations of HHT from 1 to 4 mg/ml didn't have an effect on cell viability. In RBL-2H3 cells, ${\beta}$-hexosaminidase release, IL-4 and TNF-${\alpha}$ were significantly reduced by treated all concentrations of HHT. COX-2 on mRNA expression was reduced at treatment 2mg of HHT prior to stimulating by A23187. These results indicate that HHT has inhibition effects on allergy in this study.

• Korean Journal of Acupuncture
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• v.28 no.3
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• pp.73-83
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• 2011

Objectives : The purpose of this study is to investigate anti-allergic effects of Glycyrrhizae Radix(GR) pharmacopuncture and GR extract. Methods : In vivo, animals were gotten GR pharmacopunctures at both sides of ST36s three times for 5 days. Then, we investigated anti-DNP IgE-induced passive cutaneous anaphylaxis of Sprague Dawley rats. In vitro, we measured cell viability, ${\beta}$-hexosaminidase release, and the secretion of interleukin-4(IL-4) and tumor necrosis factor-alpha(TNF-${\alpha}$) in RBL-2H3 cells after treatment of various concentrations of GR extract. Results : In vivo, we observed inhibition of passive cutaneous anaphylaxis after GR pharmacopuncture treatments at both sides of ST36s and optional points. In vitro, GR extract treatments did not affect cell viability, but inhibited ${\beta}$-hexosaminidase release and the secretion of IL-4 and TNF-${\alpha}$. Conclusions : These results suggest that GR pharmacopuncture and GR extract should be beneficial in the inhibition of allergic inflammatory response.

• The Journal of Pediatrics of Korean Medicine
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• v.26 no.1
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• pp.1-15
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• 2012

$CheongGiGeoYangTang$ has been used for anti-allergenic purpose. However there was no experimental study about its effect. Therefore, this study was designed to investigate the anti-allergenic effect of $CheongGiGeoYangTang$. Methods Modifiability of RBL-2H3 mast cells' IL-4, IL-13 was analyzed by qRT-PCR and ELISA. Also, the suppressive effect of GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-${\kappa}B$ p65 transcription factors was observed by western blotting. OVA-specific IgE, IL-4 and IL-13 production in ovalbumin allergy model was examined as well. Results It was showed that the RBL-2H3 mast cells treated with $CheongGiGeoYangTang$ extract(CGGYT) was significantly suppressed mRNA expression, production of IL-4 and IL-13, and prominently inhibited the expression of transcription factors including GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos and NF-${\kappa}B$ p65 but not c-Jun. The administration of CGGYT was suppressed the amount of OVA-specific IgE, IL-4 and IL-13 in OVA/alum-sensitized mice. Conclusions We considered CGGYT would regulate the allergic inflammation as inhibition of IL-4 and IL-13 production in activated mast cells and Th2 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.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.1
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• pp.127-134
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• 2009

In the present study, we investigated the anti-allergic effect of the water extract of Bopyeoyangyeong-jun(BYJ) to cytokines and transcription. To investigate the biological effect of BYJ, We examined cytotoxicity and inflammatory cytokine secretion with RBL-2H3. We examined tumor necrosis factor-alpha(TNF-$\alpha$), interleukin(IL)-4 secretion from RBL-2H3 cell after pre- treatment with Bopyeoyangyeong-jun of $1\;mg/m{\ell}$, $2\;mg/m{\ell}$. RBL-2H3 cell was stimulated with phorbol 12-myristate 13-acetate(PMA) and calcium ionophore A23187. We observed that Bopyeoyangyeong-jun reduced TNF-$\alpha$, IL-4 secretion and mRNA expression in RBL-2H3 cells. Moreover, the expression of levels of cyclooxygenase (COX)-2 mRNA, nuclear factor-kappa B (NF-${\kappa}B$) (p65) protein, ERK MAPK, and the degradation of level inhibitor kappa B-alpha ($I{\kappa}B-{\alpha}$) were down-regulated by BYJ. Taken together, these results indicate that Bopyeoyangyeong-jun hascontrols TNF-$\alpha$, IL-4 secretion on allergic reaction.

• The Journal of Pediatrics of Korean Medicine
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• v.26 no.4
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• pp.10-23
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• 2012

Objectives: Gagamyanggyeoksan (G-YGS) has been used to suppress allergic reaction, however, the cellular target of G-YGS and its mode of action remain unclear. The present study was designed to investigate the effect of extracted G-YGS on the PMA and lonomycin (PI)-induced activation of RBL-2H3. Methods: For this investigation, We examined IL-4, IL-13 mRNA expression by Real-Time PCR, IL-4, IL-13 production by ELISA analysis and manifestations of GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-${\kappa}B$ p65 transcription factors by western blotting, OVA-specific IgE, IL-4, IL-13 by mouse be sensitive to OVA. Results: Here we showed that treatment of RBL-2H3 mast cells with G-YGS, suppressed PI-induced production of Th2 cytokines including IL-4 and IL-13 in a dose dependent manner. The mRNA expression of IL-4 were completely abolished by G-YGS at the concentration of $100{\mu}g/ml$. Data from a stable cell lines consistently expressing IL-4. And the mRNA expression of IL-13 were abolished by G-YGS at the $200{\mu}g/ml$. But there is no difference between the $50{\mu}g/ml$, the $100{\mu}g/ml$ and the comparison. Results from the western blot analysis of transcription factors involving IL-4 and IL-13 expression indicated that it prominently decreased the expression of mast cell specific transcricption factors including GATA-1, GATA-2, NF-AT2, c-Jun, NF-${\kappa}B$ p65 but not c-Fos. And G-YGS suppressed IgE, IL-4, IL-13 in mouse be sensitive to OVA. Conclusions We suggested the anti-allergic activities of G-YGS might be mediated by down-regulation of Th2 cytokines such as IL-4 and IL-13 through the regulation of transcription factors as GATA-1, GATA-2, NF-AT2, c-Jun, NF-${\kappa}B$ p65.