• 제목/요약/키워드: MAST assay

검색결과 114건 처리시간 0.023초

Sinensetin Inhibits Interleukin-6 in Human Mast Cell - 1 Via Signal Transducers and Activators of the Transcription 3 (STAT3) and Nuclear Factor Kappa B (NF-κB) Pathways

  • Chae, Hee-Sung;Kim, Young-Mi;Chin, Young-Won
    • Natural Product Sciences
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    • 제23권1호
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    • pp.1-4
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    • 2017
  • Sinensetin, a pentamethoxyflavone, is known to exert various pharmacological activities including anti-angiogenesis, anti-diabetic and anti-inflammatory activities. However, its effects on the human mast cell - 1 (HMC-1) mediated inflammatory mechanism remain unknown. To explore the mediator and cellular inflammatory response of sinensetin, we examined its influence on phorbol 12-myristate 13-acetate (PMA) plus A23187 induced inflammatory mediator production in a human mast cell line. In this study, interleukin (IL)-6 production was measured using the enzyme-linked immunosorbent assay and reverse transcription polymerase chain reaction. Sinensetin inhibited PMA plus A23187 induced IL-6 production in a dose-dependent manner as well as IL-4, IL-5 and IL-8 mRNA expression. Furthermore, sinensetin inhibited signal transducer and activator of transcription 3 (STAT3) phosphorylation, suggesting that sinensetin inhibits the production of inflammatory mediators by blocking STAT3 phosphorylation. Moreover, sinensetin was found to inhibit nuclear factor kappa B activation. These findings suggest that sinensetin may be involved in the regulation of mast cell-mediated inflammatory responses.

머위(Petasites japonicum) 추출물의 항알레르기 효과 (Anti-allergic Effects of Petasites japonicum)

  • 최옥범
    • 한국식품영양학회지
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    • 제15권4호
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    • pp.382-385
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    • 2002
  • 본 연구에서는 민간에서 옻나무에 의 한 알레르기반응 및 기관지천식의 치료 효과가 있는 것으로 알려진 머위(Petasites japonicum)추출물의 항알레르기 효과를 RBL-2H3 세포로부터 hexosaminidase의 방풀 억제 정도를 측정하는 enzyme assay를 이용하여 검색하였다. 실험결과에 대한 오차범위가 낮은 추추출 300 $\mu\textrm{g}$/mL 농도의 수준에서 methanol 추출물은 83.33%, ethylacetate 추출물은 69.75%, 열수 추출물은 50.62%의 저해효과를 나타냈으며, 대조구로 사용한 quercetin은 같은 농도에서 85.80%의 저해 효과를 나타냈다. 따라서 정제된 quercetin의 hexosaminidase의 방출 억제 활성을 비교해 볼 때 머위 추출물이 조추출물 상태인 점을 고려한다면 상당한 항알레르기 효과가 있음을 시사하고 있어 새로운 항알레르기성 신기능 물질로의 이용방안을 탐색하는 기초자료가 되리라 사교된다.

Serum allergen-specific and total IgE concentrations in dogs with allergic disorders in South Korea

  • Byoung-Gook Yoon;Han-Joon Lee;Kun-Ho Song
    • 한국동물위생학회지
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    • 제46권3호
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    • pp.219-225
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    • 2023
  • Canine atopic dermatitis (CAD) has a genetic basis and is primarily a skin disorder characterized by allergic features. It is caused by an excessive immune response, associated with immunoglobulin E(IgE), to allergens and shows clinical signs of itching and inflammation. The present study evaluated the allergen-specific and total IgE concentration in dogs with allergic disorders. Allergen-specific IgE analysis was performed using a Multiple Allergen Simultaneous Test (MAST) assay, and total IgE analysis was conducted using an ELISA in 82 dogs with CAD. The subjects were divided into two groups: dogs aged ≤3 years and dogs aged >3 years, enabling a comparison of total IgE concentrations between the two age groups. The results showed that the major affecting environmental allergen was Alternaria alternate, and the major affecting dietary allergen was raw chestnut. The results revealed that dogs aged ≤3 years had higher total IgE concentrations than dogs aged >3 years with significant difference. The results of this study can contribute to the diagnosis and treatment of CAD in South Korea.

케어스웰을 이용한 사이토카인 억제 효과에 대한 연구 (The Study of Cytokine Inhibitory Effect using Careswell)

  • 하효철;오사랑;이지혜;정지욱
    • 대한본초학회지
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    • 제29권3호
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    • pp.79-85
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    • 2014
  • Objectives : In this study, we attempted to evaluate the effects of Careswell on human mast cell-mediated allergy inflammation in vitro and pruritogen-induced scratching behavior in vivo. Method : The Careswell was extract by distilled water. The anti-itching effects of Careswell were investigated on the compound 48/80 ($50{\mu}g/kg$) or histamine ($100{\mu}g/kg$) induced scratching behavior male ICR mice for 30 min by an observer blind. Terfenadine (10 mg/kg) was used as a positive control drug. The cell toxicity of Careswell was determined by 3-(4,5-dimethylthiazole-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay. The regulatory effect of Careswell on interleukin (IL)-6 and tumor necrosis factor (TNF)-${\alpha}$ levels was determined by enzyme-linked immunosorbent assay (ELISA) in phorbol 12-myristate 13-acetate plus calcium ionophore A23187 (PMACI) stimulated human mast cells (HMC-1). Also, we evaluated the effect of Careswell on PMACI induced the activation of Nuclear factor-kappa B (NF-${\kappa}B$) into nucleus by Western blot analysis. Result : The results revealed that the oral administration of Careswell (200 mg/kg, p.o.) attenuated the compound 48/80 or histamine-induced scratching behavior in mice. We showed that Careswell significantly reduced the PMACI-induced the production of IL-6 (0.5-1 mg/ml) and TNF-${\alpha}$ (0.1-1 mg/ml). Additionally, Careswell significantly inhibited the activation of NF-${\kappa}B$ in PMACI-stimulated HMC-1. Conclusion : Collectively, the findings of this study provide us with a novel insight into the pharmacological actions of Careswell as a potential molecule for use in the treatment of allergic inflammation diseases.

백렴의 알레르기 염증반응에 대한 억제효과 (The Effects of Ampelopsis Radix on Allergic Inflammation in PMA-stimulated Human Mast Cells)

  • 김장현;천진홍;김성윤;박용기
    • 대한본초학회지
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    • 제23권4호
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    • pp.91-101
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    • 2008
  • Objectives: In this study, we investigated the inhibitory effects of Ampelopsis Radix methanol(AR-M) extract on allergic inflammation in activated human mast cells and its potential therapeutic or toxic effects. Methods: Ampelopsis Radix(AR) was extracted with 80% methanol. HMC-1 cells, a human mast cell line, were treated with different concentrations of AR-M extract, and then stimulated with PMA plus A23187. The cell toxicity of AR-M extract was determined by MTT assay. The concentrations of $PGE_2$ and cytokines were measured by ELISA. The gene expression of COX-2 and its protein levels were determined by RT-PCR and Western blot. The phosphorylation of ERK MAPK and the NF-${\kappa}B$ activation were determined by Western blot. Results: AR-M extract was significantly inhibited the production of PGE2 and inflammatory cytokines(TNF-${\alpha}$, IL-6 and IL-8) in PMA/A23187-stimulated HMC-1 cells. AR-M extract also attenuated the mRNA expression of COX-2 and its protein induction. Furthermore, AR-M extract attenuated PMA/A23187-induced phophorylation of ERK1/2 MAPK and the NF-${\kappa}B$ p65 subunit translocation into nuclear of HMC-1 cells. AR-M extract significantly decreased PMN A23187-induced release of histamine in a dose-dependent manner. Conclusions: These results indicate that Ampelopsis Radix shows the property of anti-allergic inflammation In vitro through suppressing the production of inflammatory mediators released from mast cells, suggesting have a potential for the treatment of allergic diseases.

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RBL-2H3 비만세포에서 유백피 에탄올 추출물의 알레르기 반응 개선에 대한 효과 (Effect of Ullmus macrocarpa Hance Ethanol extract (Ulmus) on Improvement of allergic responses in RBL-2H3 mast Cells)

  • 도현주;오태우
    • 대한한의학방제학회지
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    • 제29권4호
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    • pp.191-203
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    • 2021
  • Objectives : In this study, we investigate the anti-allergic effects of Ullmus macrocarpa Hance (Ulmus) on RBL-2H3 mast cell (basophilic leukemia cell line), which are mediated by FcεRIs. Methods : We evaluated the effect of the ethanol extract of Ulmus on the allergic inflammatory response in IgE-antigen-mediated RBL-2H3 cells. Cell toxicity was determined by MTT assay and the markers of degranulation such as beta-hexosaminidase, histamine, PGD2, TNF-α, IL-4, IL-6 production of inflammatory mediators and FcεRI-mediated protein expression by western blot. Results : Ulmus inhibited degranulation and production of allergic mediators (e.g., TNF-α, IL-4, and IL-6) in them. Ulmus reduced histamine levels, expression of FcεRI signaling-related genes such as Lyn, Syk, and Fyn, and extracellular signal-regulated kinase phosphorylation in mast cells. Also, Ulmus reduced PGD2 release and cyclooxygenase-2 expression, and cytosolic phospholipase A2 phosphorylation in FcεRI-mediated RBL-2H3 mast cells. Conclusions : These results indicate that Ulmus exhibits anti-allergic activity through inhibition of degranulation and inflammatory mediators and cytokine release. These findings suggest that Ulmus may have potential as a prophylactic and therapeutic agent for the treatment of various allergic diseases.

Human mast cell에서 승마갈근탕(升麻葛根湯)의 항염증 효과에 대한 연구 (Anti-inflammatory effect of Seungmagalgeun-tang extract in human mast cells)

  • 금준호;서윤수;강옥화;최장기;권동렬
    • 대한본초학회지
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    • 제28권5호
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    • pp.7-11
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    • 2013
  • Objectives : Seungmagalgeun-tang (SMGGT) is traditional medicine widely used for inflammatory disease and flu. But SMGGT exhibits potent anti-inflammatory activity with an unknown mechanism. To elucidate the molecular mechanisms of SMGGT water extract on pharmacological and biochemical actions in inflammation, we examined the effect of SMGGT on pro-inflammatory mediators in Phorbol-12-myristate-13-acetate (PMA)+A23187-stimulated mast cells. Methods : In the present study, pro-inflammatory cytokine production was determined by performing enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and western blot analysis to measure the activation of MAPKs. Cells were treated with SMGGT 1 h prior to the addition of 50 nM of PMA and $1{\mu}M$ of A23187. Cell viability was measured by MTS assay. The investigation focused on whether SMGGT inhibited the expressions of interleukin-6 (IL-6), interleukin-8 (IL-8) and mitogen-activated protein kinases (MAPKs) in PMA+A23187-stimulated mast cells. Results : SMGGT has no cytotoxicity at examined concentration (100, 250, and $500{\mu}g/ml$). Also, gene expression of IL-6 and IL-8 in HMC-1 cells stimulated by PMA+A23187 was down regulated by SMGGT. Furthermore, SMGGT suppressed the PMA+A23187-induced phosphorylation of extracellular signal-regulated kinase (ERK) and c-jun N-terminal Kinase(JNK). But, SMGGT could not regulate phosphorylation of p38 MAPK. Conclusions : These results suggest that SMGGT has inhibitory effects on PMA+A23187-induced IL-6 and IL-8 production. These inhibitory effects occur through blockades on the phosphorylation of ERK and JNK.

Inhibitory Effect of Alpiniae officinarum Rhizoma Extract on Degranulation in RBL-2H3 Cells

  • Kim, Eunhee;Ahn, Sejin;Lee, Deug-Chan
    • 한국자원식물학회지
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    • 제28권3호
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    • pp.321-328
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    • 2015
  • Alpiniae officinarum Rhizoma (the rhizome of Alpinia officinarum Hance, known as lesser galangal), a family of Zingiberaceae, has been used to reduce pain of infection and inflammatory diseases in Asian countries. The present study was focused to evaluate the inhibitory degranulation effect of Alpiniae officinarum Rhizoma extract in RBL-2H3 rat basophilic leukemia cells. Cell viability was measured by MTT assay. RBL-2H3 cells were stimulated by phorbol 12-myristate 13-acetate and calcium ionophore A23187. Mast cell degranulation was analyzed by measuring release of β-hexosaminidase in RBL-2H3 cell. Gene expression was measured by qRT-PCR and signaling molecules were detected by immunoblotting. The Alpiniae officinarum Rhizoma extract suppressed β-hexosaminidase release in dose-dependent manner and inhibited cycloxygenase-2 and tumor necrosis factor-α gene expression. Furthermore, it was found that Alpiniae officinarum Rhizoma extract reduced mitogen-activated protein kinases, especially phosphorylated p38, at 0.75 ㎎/㎖ of Alpiniae officinarum Rhizoma extract concentrations. These data show that Alpiniae officinarum Rhizoma extract has immunosuppressive effect in mast cell induced allergic inflammation.

감잎(Diospyros kaki folium) Polyphenol 화합물군이 알레르기성 접촉피부염에 미치는 영향 (Effect of Polyphenol Compounds from Persimmon Leaves (Diospyros kaki folium) on Allergic Contact Dermatitis)

  • 박무희;최청;배만종
    • 한국식품영양과학회지
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    • 제29권1호
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    • pp.111-115
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    • 2000
  • 한국산 감잎으로부터 폴리페놀 화합물군을 분리하여 알레르기성 접촉피부염에 미치는 영향을 관찰하였다. 그 결과 대조군에 비해 폴리페놀 화합물군에서 era swelling이 감소되었으며, 비만세포의 분포에서도 대조군에 비하여 감잎 폴리페놀 화합물 투여군에서 매우 적은 수가 확인되었으며, 이러한 결과는 제4형 알레르기 반응을 지연 또는 예방할 수 있는 생약제로서의 가능성을 시시하는 것으로 보여진다.

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흰쥐 복강 비만세포에서 Dendroaspis natriuretic peptide에 의한 히스타민 유리 (HISTAMINE RELEASE INDUCED BY DENDROASPIS NATRIURETIC PEPTIDE FROM RAT PERITONEAL MAST CELLS)

  • 김재곤;허선;백병주
    • 대한소아치과학회지
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    • 제28권1호
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    • pp.72-81
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    • 2001
  • Dendroaspis natriuretic peptide (DNP)는 최근에 green Mamba snake Dendroaspis angusticeps의 독(venom)에서 발견된 17개의 아미노산 disulfide링 구조를 포함하는 38개 아미노산 펩티드로 natriuretic peptide family와 그 구조가 비슷하다. Natriuretic peptide family는 사람과 흰쥐 비만세포로부터 히스타민을 유리시킨다고 알려져 있다. 그러나 DNP가 비만 세포로부터 히스타민을 유리 시키는 여부는 밝혀져 있지 않다. 본 연구는 DNP가 흰쥐 복강 비만세포로부터 히스타민을 유리시키는 여부와 함께 히스타민 유리기전을 구명하고자 실시되었다. 다양한 농도의 DNP를 흰쥐 복강 비만세포에 처리한 다음, 비만세포의 탈과립을 도립 광학현미경으로 관찰하였다. 히스타민 유리량은 방사효소법으로 측정하였고 세포내 칼슘 유입량과 cyclic GMP의 수준은 방사면역법으로 측정하였다. DNP는 흰쥐 복강 비만세포를 탈과립시켰고 농도가 증가함에 따라 비만세포로부터 히스타민 유리량이 증가되었다. 또한 DNP는 농도 증가에 비례하여 비만세포 안으로 세포밖의 칼슘을 유입시켰으며 세포안의 cyclic GMP의 수준을 증가시켰다. 이상의 결과로 미루어, DNP는 비만세포 안의 cyclic GMP와 칼슘의 농도를 증가시켜 비만세포로부터 히스타민을 유리시키는 것으로 생각된다.

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