• Title/Summary/Keyword: MAST assay

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Effects of Tribuli Fructus extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells (비만세포에서 백질려 추출물의 항염증효과에 대한 연구)

  • Rho, Hyo Sun;Park, Yong-Ki;Bae, Hyo Sang
    • The Korea Journal of Herbology
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    • v.32 no.2
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    • pp.107-114
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    • 2017
  • Objectives : Tribulus terrestris $Linn{\acute{e}}$ (Tribuli Fructus; TF) has been used to treat hypochondrium, agalactia, nebula, itching and vitiligo in traditional Korean medicine. In this study, we investigated the effects of TF 30% ethanol extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells. Methods : TF extract was prepared by 30% ethanol. RBL-2H3 cells, a rat mast cell line, were treated with TF extract at different concentrations for 1 hr and then stimulated with DNP-IgE/HSA for indicated times. Cell viability was measured by WST-1 assay. The expression of inflammatory cytokines (IL-4, IL-13 and $IFN-{\gamma}$) mRNA was determined by reverse transcriptase-PCR, and the phosphorylation of ERK1/2, p38 and JNK MAP kinases (MAPKs) was determined by Western blot. The nuclear expression of $NF-{\kappa}B$ p65 in the cells was detected by Western blot and immunocytochemistry, respectively. Results : The treatment of TF extract at 0.1 and $0.2mg/m{\ell}$ significantly decreased the expression of IL-4 and IL-13 mRNA in IgE-stimulated RBL-2H3 mast cells, while significantly increased the expression of $IFN-{\gamma}$ mRNA. TF extract treatment was also inhibited the phosphorylation of ERK1/2, p38 and JNK MAPKs in IgE-stimulated RBL-2H3 mast cells in a dose-dependent manner. In addition, TF extract significantly blocked the translocation of $NF-{\kappa}B$ p65 into the nuclear of cells after IgE stimulation. Conclusions : These results indicate that TF extract inhibits inflammatory response in IgE-stimulated mast cells through blocking MAPKs/$NF-{\kappa}B$ pathway. This suggests that TF extract has an anti-inflammatory activity in mast cell activation.

The Effect of Yunkyopaedocksangamibang on Allergic Contact Dermatitis : Based on the mitigation of skin damage and distributive change of mast cell on DNCB re-expasure region of Mice (연교패독산가미방(連翹敗毒散可味方)이 알러지성 접촉피부염에 미치는 영향(影響) - DNCB 재노출부위의 생쥐 피부손상완화와 비만세포 분포 변화를 중심으로 -)

  • Kwon, Oh-Sung;Kim, Jin-Tack;Park, In-Sick;Ahn, Sang-Hyun;Lee, Hai-Poong;Kim, Ho-Hyun;Gang, Yun-Ho
    • The Journal of Dong Guk Oriental Medicine
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    • v.8 no.1
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    • pp.77-91
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    • 1999
  • After allergic contact dermatitis elicitated by Dinitrochlorobenzene(DNCB) treatment, ICR female mice administered Yunkyopaedocksangamibang(YPGM) extract were observed to investigate the effect of YPGM on allergic contact dermatitis. This study investigated that contact hypersensitivity assay, abdominal skin morphologic changes including mast cells. At contact hypersensitivity assay, the right ear swelling in YPGM group were probability decreased than DNCB group. At observation of abdominal skin morphologic change, the infiltration of lymphocyte, lymphocyte insertion to epithelium, enlarged capillary, angiogenesis, and damages of epithelium as cytoplasmic vacuolation and enlarge of inter cellular space in YPGM were diminished than DNCB group. The number of mast cell was increased both DNCB and YPGM group. The shape of mast cell in DNCB group was mainly appeared degranulated type, but granulated type in YPGM group. The number of serotonin positive cell was increased both DNCB and YPGM group. The shape of serotonin positive cell in DNCB group was mainly appeared degranulated type, but granulated type in YPGM group. As results indicated that the YPGM extract administration work on the mitigation of skin damages in mouse with allergic contact dermatitis.

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Correlation of inhalant MAST on chronically coughing children (만성 기침을 하는 소아에서 호흡기 증상과 inhalant MAST의 연관성)

  • Yun, Jung Min;Roh, Eui Jung;Lim, Jae Woo;Choen, Eun Jung;Ko, Kyoung Og;Lee, Young Hyuk
    • Clinical and Experimental Pediatrics
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    • v.49 no.6
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    • pp.643-647
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    • 2006
  • Purpose : Recently, incidences of chronic cough with allergic disease have been increased. The identification of specific allergens for chronically coughing children is significant for diagnosis, proper treatment and prevention. The aim of this study was to identify significant contributors for sensitization in chronically coughing children. Methods : A total of 106 children, aged 1-6 years, visiting for chronic cough(>3 weeks), were tested for inhalant multiple allergosorbent assay(MAST), eosinophil count and total IgE. Parents completed a questionnaire on past history of atopy(infantile eczema) and bronchiolitis, family history of allergy, breastfeeding, smoking, pets and floor coverings at home. Results : Significant contributors for sensitization were : age(4-6 years), infantile eczema, positive family history of allergy and smoking by family. Conclusion : We were able to identify significant contributors for sensitization in chronically coughing children.

The Effects of Ethanol Extract from Atractylodes Chinensis Rhizome on the Mast Cell-Mediated Inflammatory Responses (창출(蒼朮) 에탄올 추출물이 비만세포 매개 염증반응에 미치는 영향)

  • Kim, Sun-Min;Kim, Kyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.24 no.1
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    • pp.45-63
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    • 2011
  • Objective : Atractyloides Chinensis Rhizome (ACR) is widely used in oriental medicine as a remedy for an inflammation and an allergic disease. However, as yet there is no clear explanation of how ACR affects the production of inflammatory cytokine. This study was to determine the effects of ACR on the mast cell-mediated inflammatory responses. Method : The amount of inflammatory cytokine production induced by the phorbol myristate acetate (PMA) plus calcium ionophore(A23187) in the human mast cell line (HMC-1) incubated with various concentrations of ACR was measured. The TNF-${\alpha}$ protein levels were analysised by Western blots. The TNF-${\alpha}$, IL-6 and IL-8 secreted protein levels were measured by the ELISA assay. The TNF-${\alpha}$, IL-6 and IL-8 mRNA levels were measured by the RT-PCR analysis. NF-${\kappa}$B, phospho-I${\kappa}$B and MAPKs were examined by Western blot analysis. The NF-${\kappa}$B promoter activity was examined by a luciferase assay. Results : 1. The expressions of TNF-${\alpha}$ and TNF-${\alpha}$ mRNA were decreased dose-dependently at 0.05-0.2mg/$m\ell$ of ACR and significantly decreased at 0.2mg/$m\ell$. 2. The expressions of IL-6 and IL-6 mRNA were decreased dose-dependently at 0.05-0.2mg/$m\ell$ of ACR and significantly decreased at 0.2mg/$m\ell$. 3. The expressions of IL-8 and IL-8 mRNA were decreased dose-dependently at 0.05-0.2mg/$m\ell$ of ACR and significantly decreased at 0.2mg/$m\ell$ specially. 4. The expressions of Phosphorylated-JNK were decreased, not p38, ERK 5. The expressions of NF-${\kappa}$B were decreased dose-dependently at 0.1-0.2mg/$m\ell$ of ACR. The expressions of Phosphorylated I${\kappa}$B were significantly decreased at 0.2mg/$m\ell$. In addition, ACR suppressed PMA plus A23187-induced NF-${\kappa}$B promoting activity. Conclusion : It is suggested that ACR should suppress through inhibition of NF-${\kappa}$B activity and cytokine production.

Anti-allergic Effect of Gyokejsamu1tang-gagam(艽桂四物湯-加減) in the RBL-2H3 Mast Cells (RBL-2H3 cells에서 GyoKeisamultang-gagam의 항알레르기 효과)

  • Tsung, Pei-Yun;Shin, Woo-Jin;Lyu, Ji-Hyo;Kim, Seong-Hui;Yoon, Hwa-Jung;Kim, Won-Il
    • 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.

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Anti-allergic Effects of Castanea crenata Leaf Tea (밤나무 잎차(茶)의 항알레르기 효과)

  • Choi, Ok-Beom;Kim, Kyung-Man;Yoo, Gyurng-Soo;Park, Keun-Hyung
    • Korean Journal of Food Science and Technology
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    • v.30 no.2
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    • pp.468-471
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    • 1998
  • Regarding the characterstics of allergic diseases, preventive and continuous treatment is desirable, and tea would be the one of the best functional food formula for it. Here we report the development of tea processing method for the leaves of Castnaea crenata. Two forms of Castnaea crenata leaf tea were prepared, non-fermented steaming tea and semi-fermented rolling tea. Anti-allergic actions of Castanea crenata leaf tea were asessed by testing their effects on the degranulation of mast cells. For this, hexosaminidase release (degranulation marker) from RBL-2H3 cells (mast cell line) was used. At the concentration of $300\;{\mu}g/mL$ of the water extract, the degranulation of RBL-2H3 cells were inhibited 50.4% and 35.4% by non-fermented steaming tea and semi-fermented rolling tea, respectively. These results suggest that the tea processing method we developed could provide a valuable resource for the treatment of allergic diseases.

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Anti-Inflammatory Effects of Bee Venom on Phthalic Anhydride-Induced Atopic Dermatitis

  • Oh, Myung Jin;Song, Ho-Sueb
    • Journal of Acupuncture Research
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    • v.37 no.1
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    • pp.42-48
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    • 2020
  • Background: Atopic dermatitis (AD) is a chronic inflammatory condition which can be studied using phthalic anhydride (PA) to induce AD. Anti-inflammatory properties of bee venom (BV) wereinvestigated to determine whether it may be a useful treatment for AD. Methods: AD was induced by applying to pical PA to 8-week-old HR-1 mice (N = 50), then treating with (0.1, 0.25, and 0.5 ㎍) or without topical BV. Body weight, ear thickness histology, enzyme-linked immune sorbent assay (serum IgE concentrations), Western blot analysis [inducible nitric oxide synthase, cyclooxygenase-2, IκB-α, phospho-IκB-α, c-Jun N-terminal kinase (JNK), phospho-JNK, p38, phospho-p38, extra cellular signal-regulated kinase (ERK), and phospho-ERK], and the pull down assay for immunoblotting (p50), were used to measure inflammatory mediators. Results: PA + BV (0.1, 0.25, and 0.5 ㎍) significantly decreased ear thickness without altering body weight. IgE concentrations decreased in the PA + BV (0.5 ㎍)-treated groups compared with PAtreatment. Tumor necrosis factor-α, interleukin-1β, inducible nitric oxide synthase, cyclooxygenase-2, phospho-IκB-α, phospho-JNK, p38, phospho-p38, and phospho-ERK, all decreased following treatment with PA + BV compared with the PA-treatment alone. p50 was upregulated in the PA + BV-treated groups compared with the PA-treated group. Furthermore, the number of mast cells decreased in the PA + BV-treated groups compared with the PA-treated group. Epidermal thickness was significantly lower in the PA + BV-treated group compared with PA treatment alone. Conclusion: BV maybe a useful anti-inflammatory treatment for AD.

Inhibitory Effect of Inflammatory Cytokines Secretion of Cheonggeumganghwa-tang in Mast cell (비만세포에서의 청금강화탕의 염증성 세포활성물질 분비 억제 효과)

  • Choi Young Soo;Moon Goo;Kim Dong Woung;Han Se Hee;Won Jin Hee
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.3
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    • pp.887-892
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    • 2004
  • Cheonggeumganghwa-tang(CGT) has been used for the purpose of prevention and treatment of bronchial asthma and allergic asthma in Korea. To investigate the biological effect of CGT, the author examined cytotoxicity and inflammatory cytokines secretion with human mast cell line, HMC-1. HMC-1 was stimulated with phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187. CGT by itself had no effect on viability of HMC-1. The effects of CGT on the secretion of tumor necrosis factor-alpha (TNF-α) and interleukin (IL)-6 from HMC-1 were evaluated with enzyme-linked immunosorbent assay (ELISA). CGT (1 ㎎/㎖) inhibited PMA plus A23187 -induced TNF-α and IL-6 secretion, by 93.86 ± 2.05%, 68.69 ± 2.86%, respectively. CGT also inhibited the NF-κB (p50) expression. Taken together, these results suggest that CGT inhibit the production of inflammatory cytokines in HMC-1 cells through blockade of NF-κB activation.

Research on Allergy-Causing Materials of Allergy Diseases (알레르기 의심환자의 알러겐 발생빈도에 대한 연구)

  • Kim, Yoon-Sik;Kim, Yeon
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.3
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    • pp.201-209
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    • 2007
  • Among the inhalant and foods which caused allergy, which is increasing nowaday, this research has a purpose of studying allergy-causing materials (allergen) which most frequently cause allergy. In order to conduct this research, we selected 100 hundred patients (M : F = 65 : 35) from in Kwangju Christian Hospital, who were tested through allergy multiple antigen simultaneous test (MAST) immunoblot assay (RIDA$^{(R)}$ X-Screen; r-Biopharm Co., Darmstadt, Germany). According to the test, the total IgE positive rate (above Class 2) from the inhalant is 58.3%, and that from food is 35%, respectively. This research showed that the first most allergen was the tick (Derm. farinae/Derm. pteronyssinus), the second most was housedust, and the third was the hair of pets such as cats and dogs; 65%, 35%, 26.3% in inhalant panel, and 52.5%, 40%, 50% in food panel, respectively. Thease allergens newly added to inhalant and food panels were not contributable to the detection of specifical allergen. We needs further improvement to be used as primary screening or quantitative test for allergy.

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Effects of Panax ginseng on Type I Hypersensitivity (제1형 과민 반응에 미치는 고려인삼의 영향)

  • Kim, Young-Ran;Lee, Eun;Lee, Shee-Yong;Kim, Kyeong-Man
    • Journal of Ginseng Research
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    • v.20 no.1
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    • pp.1-6
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    • 1996
  • Effects of Panax ginseng on allergic reactions were studied uslng various in vivo and in vitro experimental models such as 48-hr passive cutaneous anaphylaxis, mediators-induced skin reactions, histamine release from rat peritoneal mast cells, hexosaminidase release from RBL-2H3 cells, and lipoxygenase assay . In all of anti-allergic experiments we conducted, ginseng components (50% ethanol extract or ginseng total saponin or ginsenosides) extracted from Korean red ginseng, did not show significant anti-allergic actions. In 48-hr passive cutaneous anaphylaxis and mediators-induced skin reactions, 50% ethanol extract did not suppress hypersensitivity reactions. Total saponin, 50% ethanol extract, and 8 major ginsenosides did not show inhibitory effects on lipoxygeanse activity. Ginseng total saponin did not inhibit histamine release from rat peritoneal mast cells. All of the ginseng components mentioned above were also tested on RBL-2H3 cells, but none of them inhibited hexosaminidase release from this cell line. These results suggest that Panax ginseng does not have effects on allergic reactions at the level of 50% ethanol extract or total saponin used. All of 8 major saponin components tested ($Rb_1$, $Rb_2$, Rc, Rd, Re, Rf, $Rg_1$, $Rg_2$), did not inhibit lipoxygenase activity and degranulation events.

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