• 디지털융복합연구
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• 제13권8호
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• pp.503-514
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• 2015

은행잎 유래 플라보놀 성분이 갖는 항아토피 활성에 대해 입증된 바는 드물다. 이 논문에서는 MC/9 비만세포에서의 은행잎 플라보놀의 항아토피 효과를 조사하기 위하여 ELISA와 Real-time PCR, western blot으로 은행잎 플라보놀을 분석하였다. 분석 결과 IL-13, MIP-1a의 생성량은 농도 의존적으로 현저하게 감소되었으며 IL-4, IL-5, IL-13, TNF-a 유전자 발현량은 25, 50, $100{\mu}g/m{\ell}$의 농도에서 효과적으로 억제되었다. western blot 분석 결과 c-jun과 NFAT-1단백질 발현이 감소되었음을 확인하였다. 이러한 결과들은 은행잎 유래 플라보놀 성분이 NFAT-1, c-jun 전사인자의 전사를 억제함으로써 MC/9 비만세포에서의 Th2 싸이토카인의 생성을 감소시키는 효과를 갖는 것을 나타낸다. 따라서 은행잎 유래 플라보놀 성분이 아토피 피부염 치료제로서 활용가능성이 있음을 보고하고자 한다.

• 생명과학회지
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• 제18권10호
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• pp.1348-1354
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• 2008

mi transcription factor (MITF)는 비만세포의 분화를 조절하는 중요한 전사인자이다. 특히 MITF는 결합조직형 비만세포에서 일반적으로 발현하는 비만세포 특이적 세린 단백분해효소의 일종인 mMCP-6 유전자의 전사를 조절한다. 본 연구는 마우스 골수유래 배양비만세포에서 mMCP-6 유전자의 전사를 조절하는 MITF이형체를 규명하였다. MITF 이형체들의 발현은 RT-PCR로 확인하였다. IL-3존재 하에서 배양한 점막형 비만세포들은 MITF-A,-E, -H, -Mc 등이 발현하였다. 반면에 SCF존재 하에서 배양한 결합조직형 비만세포들은 MITF-A가 발현하였다. MITF이형체를 과발현시키면 NIH-3T3 세포에서 mMCP-6 promoter를 통한 luciferase 활성을 증가시키고, MC/9 비만세포주에서는 증가된 mMCP-6발현을 유도하였다. 더불어 비만세포에서의 mMCP-6 발현은 MITF-A 고갈로 인하여 유의적으로 억제되었다. MITF-A의 전사활성과 DNA결합은 MITF-E, -H, -Mc 등의 타 이형체들의 결과와 유사하였다. 따라서 본 연구의 결과들은 MITF-A가 마우스 결합조직형 비만세포에서 발현하여 mMCP-6 전사를 조절하는 중요한 이형체임을 제시한다.

• 대한본초학회지
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• 제28권1호
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• pp.33-42
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• 2013

Objective : Morus alba Linne Root Bark (MRAL) is a medicinal herb in Korean Medicine, known for its anti-inflammatory and anti-allergic properties. However, its mechanisms of action and the cellular targets have not yet been found and the study was developed to investigate the allergic suppressive effect of MRAL. The purpose of this study is to investigate the allergic suppressive effects of MRAL on activation of MC/9 mast cells. Methods : Cytotoxic activity of MRAL (50, 100, 200, 400 ${\mu}g/mL$) on MC/9 mast cells measured using EZ-Cytox cell viability assay kit (WST reagent). The levels of interleukin-5 (IL-5), IL-13 and IL-4, IL-5, IL-6, IL-13 mRNA expression were measured by enzyme-linked immunosorbent assay (ELISA) and real-time PCR respectively. The expression of transcription factors such as GATA-1, GATA-2, NFAT, AP-1 and NF-${\kappa}B$ p65 DNA binding activity were measured by western blot and electrophoresis mobility shift assay (EMSA). Results : Our results indicated that MRAL (50 ${\mu}g/mL$, 100 ${\mu}g/mL$) significantly inhibited PMA/Ionomycin-induced production of IL-5 and IL-13 and the expression of IL-4, IL-5, IL-6 and IL-13 mRNA in MC/9 mast cells. Moreover, MRAL (50 ${\mu}g/mL$, 100 ${\mu}g/mL$) inhibited PMA/Ionomycin-induced GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos protein expression and NF-${\kappa}B$ p65 DNA binding activity in MC/9 mast cells. Conclusions : In conclusion, we suspect the anti-allergenic activities of MRAL, may be related to the regulation of transcription factors GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos and NF-${\kappa}B$ p65 DNA binding assay causing inhibition of Th2 cytokines IL-5 and IL-13 in mast cells.

• 대한한방소아과학회지
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• 제28권2호
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• pp.23-39
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• 2014

Objectives PRAL (Prunus armniaca Linne Var) is a herbal formula in Oriental Medicine, known for its anti-inflammatory and anti-allergenic properties. However, its mechanism of action and the cellular targets have not yet been found enough. The purpose of this study is to investigate the effects of PRAL on Th2 cytokines expression in MC/9 mast cells. Methods The effect of PRAL was analyzed by ELISA, Real-time PCR, Western blot in MC/9 mast cells. mRNA levels of GM-CSF, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ were analyzed with Real-time PCR. Levels of IL-13, MIP-$1{\alpha}$ were measured using enzyme-linked immunosorbent assays (ELISA). NFAT, AP-1 and NF-${\kappa}B$ p65 were examined by Western blot analysis. Results PRAL inhibited GM-CSF, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ mRNA expression in a dose dependent manner. GM-CSF, IL-4, IL-5 mRNA expression were inhibited significantly in comparison to DNP-IgE control group at concentration of 100 ${\mu}g/ml$ and IL-6, IL-13, TNF-${\alpha}$ mRNA expression were inhibited at concentration of 50 ${\mu}g/ml$, 100 ${\mu}g/ml$. PRAL also inhibited the IL-13, MIP-$1{\alpha}$ production significantly in comparison to DNP-IgE control group in a dose dependent manner. IL-13 production was inhibited at a concentration of 200 ${\mu}g/ml$, 400 ${\mu}g/ml$ and MIP-$1{\alpha}$ was inhibited at a concentration of 100 ${\mu}g/ml$, 200 ${\mu}g/ml$, 400 ${\mu}g/ml$. Western blot analysis of transcription factors involving Th2 cytokines expression revealed prominent decrease of the mast cell specific transcription factors including NFAT-1, c-Jun as well as NF-${\kappa}B$ p65 but not NFAT-2 and c-Fos. Conclusion These results indicate that PRAL has the effect of suppressing Th2 cytokines production in the MC/9 mast cells. These data represent that PRAL potentiates therapeutic activities to the allergic disease by regulating Th2 cytokines in the MC/9 mast cells.

• 대한한방소아과학회지
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• 제28권3호
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• pp.31-46
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• 2014

Objectives Forsythiae Fructus treatment has been used for inflammatory and allergic diseases in Korean Medicine. Nevertheless, the mechanism of action and the cellular targets are not understood well. The pathogenesis of allergic diseases are associated with Th2 cytokines such as IL-13, MIP-$1{\alpha}$, IL-13, IL-5, GM-CSF, IL-4, TNF-${\alpha}$ and IL-6, which are secreted by the mast cells. This study was conducted to investigate the effects of Forsythiae Fructus extracts (FF) on Th2 cytokines expression and signal transduction in MC/9 mast cells. Methods In the study, MC/9 mast cells were stimulated with DNP-IgE for 24 hours and then treated separately with CsA $10{\mu}g/m{\ell}$ and varying doses of FF for one hour. MC/9 mast cells stimulated with DNP-IgE was the control group, a treatment with CsA was the positive control group and a treatment with varying doses FF was the experimental groups. The mRNA levels of IL-13, IL-5, GM-CSF, IL-4, TNF-${\alpha}$, IL-6 were analyzed with Real-time PCR. The levels of IL-13, MIP-$1{\alpha}$ were measured using enzyme-linked immunosorbent assays(ELISA). NFAT, AP-1 and NF-${\kappa}B$ p65 were examined by Western blot analysis. Results 1. FF were observed to suppress the mRNA expression of IL-13, IL-5, GM-CSF, IL-4, TNF-${\alpha}$, IL-6 in comparison to DNP-IgE control group. 2. FF also has inhibited the IL-13, MIP-$1{\alpha}$ production significantly in comparison to DNP-IgE control group. 3. Western blot analysis of transduction factors involving Th2 cytokines expression has revealed a prominent decrease of the mast cell specific transduction factors including NFAT-1, NFAT-2, c-Jun, and NF-${\kappa}B$ p65 but c-Fos. Conclusions In conclusion, the anti-allergenic activities of FF may be strongly related to the regulation of transcription factors NFAT-1, NFAT-2, c-Jun, and NF-${\kappa}B$ p65 causing inhibition of Th2 cytokines in mast cells.

• 대한화장품학회지
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• 제43권2호
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• pp.103-114
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• 2017

떡쑥(Gnaphalium affine D. Don, GA)은 동아시아 지역에서 식용으로 사용되고 있으며, 예로부터 전통적인 민간요법 약재로 사용되어 왔다. 현재 떡쑥 추출물(GA extract, GAE)의 항산화 활성과 항보체 활성 등은 알려져 있으나, 항염과 항알러지 효능 및 그 작용 기작은 자세히 알려져 있지 않다. 본 연구에서는 염증 매개인자인 산화질소, 프로스타글란딘 $E_2$, Toll-유사수용체 4, 에오탁신-1, 히스타민의 활성화에 대한 GAE의 저해효과를 평가하였다. 본 연구를 통해, GAE는 유도성 산화질소 합성효소와 COX-2의 발현을 저해함을 확인하였으며, 이를 통해 산화질소와 프로스타글란딘 $E_2$의 생성을 저해함을 확인하였다. GAE는 LPS로부터 유도된 Toll-유사수용체 4의 발현에도 영향을 미치는 것을 확인하였으며, A23187로부터 유도되는 비만세포의 히스타민 방출의 억제에도 효과적으로 작용하는 것을 확인하였다. 또한 IL-4로부터 유도된 에오탁신-1의 생성도 효과적으로 억제하는 결과를 확인하였다. 이상의 결과로부터 GAE는 항염증과 항알러지 효능을 가진다고 사료되며, 향후 항염증 및 항알러지 화장품 원료로서의 이용가능성을 보였다.