• Title/Summary/Keyword: MAP 인산화 효소

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Anti-inflammatory and Anti-allergic Effects of Herbal Extracts on Atopic Dermatitis ( Part II ) (약용식물 추출물의 아토피성 피부염에 대한 항염증 및 항알레르기 효과 (제 2 보))

  • Rang, Moon-Jeong
    • Journal of the Korean Applied Science and Technology
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    • v.30 no.1
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    • pp.173-182
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    • 2013
  • Atopic dermatitis is a chronic, relapsing inflammatory skin disease associated with dysfunction of skin barrier and cutaneous hyper-reactivity to environmental triggers. In the previous study, cytotoxicity, antioxidant, anti-inflammatory and anti-allergic activities were investigated for various herbal extracts such as Aloe vera L. (AV), Viola mandshurica W. Becker (VM), Punica granatum L. (PG), and Dendrobium nobile L. (DN) in order to develop effective therapeutic herbal extracts for atopic dermatitis, In this study, anti-inflammatory activities of these herb extracts in lipopolysaccharide (LPS)-induced macrophage RAW264.7 cells were further examined to find the underlying molecular mechanisms. The RT-PCR (reverse transcription polymerase chain reaction) analysis showed that PG, DN and AV inhibited effectively the gene expression of pro-inflammatory cytokines IL-6 and IL-$1{\beta}$ in LPS-stimulated macrophages, while VM did not. The transfection and luciferase analysis exhibited that all herbal extracts hindered the activation of transcription nuclear factor kappa B (NF-${\kappa}B$). The western blot analysis indicated that AV blocked the activation of only JNK MAP (c-Jun N-terminal kinase mitogen-activated protein) kinase not p38 MAP kinase, while VM, PG and DN did not show the activation of both JNK and p38 MAP kinases. These results suggest that AV, VM, PG, and DN have anti-inflammatory activities and thus have the potential to reduce and alleviate the symptoms of atopic dermatitis.

인슐린의 신호전달 기전 : Transcription Factor AP-1 의 역활

  • 김성진
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.10a
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    • pp.17-21
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    • 1995
  • 대부분의 인슐린의 작용들은 인슐린 수용체를 통하여 이루어진다. 인슐린이 수용체에 결합하면, 수용체 고유의 tyrosine kinase 효소활성의 증가를 유발시키며, 결과적으로 세포내에 존재하는 기질 단백질, IRS-1, 의 tyrosine 잔기의 인산화를 증가시키게 된다. 이후, 여러 형태의 serine / threonine protein kinase 의 연속적인 활성화가 일어난다. 이들에 부가해서, 인슐린의 효자는 세포핵 내에까지 전달되어 유전자 발현의 조절과 같은 세포핵 고유의 활동에도 관여한다. 현재, 세포막에서 시작된 인슐린의 신호들이 세포핵까지 전달되는 정확한 기전에 대해서는 알려진 바 없지만, 최근의 연구에 의하면 MAP Kinase 와 S6 Kinase 그리고 Transcription Factor AP-1의 중요성이 제시되고 있다. 특히 유전자 조절 기전에는 핵단백질인 transcription factor의 인산화 반응이 큰 역할을 한다고 보고되고 있는바, 본 연구에서 AP-1. transcription factor 의 인산화 반응이 인슐린의 신호전달계에 미치는 역할에 대하여 고찰하였다. 요약하면, AP-1 transcription factor의 구성원인 c-Jun, c-Fos 그리고 Fos 관련 단백질들의 인산화가 인슐린에 의해 증가되며, 동시에 그들의. DNA-binding activity 와 유전자 발현의 활성이 증가됨을 밝힘으로써, AP-1 transcription factor의 인산화 반응이 인슐린의 핵 내에서의 작용기전에 중요한 역할을 함이 제시되고 있다. 또한 AP-1 의 인산화 반응에 관여하는 세포핵 protein kinase로서 Casein Kinase II 의 중요성이 밝혀졌다.

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Purification and Characterization of Mitogen -Activated Protein (MAP) Kinase from Mammalian Tissue Cells (동물 조직세포로부터 Mitogen-activated Protein (MAP) Kinase의 분리 및 성격규명)

  • 김태우;정동주;김윤석
    • Biomedical Science Letters
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    • v.2 no.1
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    • pp.21-30
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    • 1996
  • MAP kinases are a family of serine/threonine specific protein kinases becoming activated in response to different proliferative stimuli by phosphorylation at both threonine and tyrosine residue. Present study shows that MAP kinase was purified from P388 murine leukema cells by SP sephadex C-50, phenyl superose and Mono Q column chromatography and identified with anti-ERKl antibody by western blotting. Immnublotting analysis to the crude extract of P388 cell lysate shows 44 kD and other minor bands but partial purified fraction eluted from phenyl supherose column have 44kD and 66 kD isoform. Subcloned GST-fusion protein from N-terminal of $p56^{kk}$ was tested as a substrate for MAP kinase phosphorylation. It was showed that the wild type and mutant forms(S42A) were fully phosporylated by purified MAP kinase fraction as com-pare with the other mutant form(S59A). This finding suggest that those GST-fusion proteins may be used as substrate for the in vitro test of MAP kinase.

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Potent Whitening Activity of Aruncus dioicus Extract in B16F10 Melanoma Cell by Suppression of Melanin Biosynthesis (흑색종세포의 멜라닌 생성억제로 인한 삼나물 추출물(Aruncus dioicus)의 미백효과)

  • Kim, Dong-Hee;Moon, Yong-Sun;Park, Tae-Soon;Hwang, Ju-Young;Son, Jun-Ho
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.813-820
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    • 2013
  • Monoterpenoids were recently found as main biologically active compounds which is responsible for various physiological effect in goat's beard (Aruncus dioicus). Ethyl acetate extract of A. dioicus (ADE) was treated to B16F10 melanoma cells for the examination of whitening activity. MTT assay was performed to evaluate cell toxicity and the result showed that slight cell toxicity (> 10%) by over $500{\mu}g{\cdot}mL^{-1}$. Thus, 0, 5, 10, or $50{\mu}g{\cdot}mL^{-1}$ ADE was used for further experiments. We found that tyrosinase activity was decreased according to ADE concentration, and the total melanin content was also dramatically reduced. Especially with $50{\mu}g{\cdot}mL^{-1}$ ADE treatment tyrosinase activity was reduced to 35.6%, and 58.8% of melanin content was lowered. In addition, whitening related proteins including tyrosinase, tyrosinase related protein 1 (TRP1), TRP2, microphthalmia associated transcription factor (MITF) and cAMP and protein kinase A (PKA) were reduced by ADE treatment. It caused decreased phosphorylation of cAMP response binding protein (CREB) but increased phosphorylation of extracellular signal related kinase (ERK). Therefore, in this paper we would like to suggest the potent usage of A. dioicus natively grown in Ulleungdo, Korea as materials of functional cosmetics by confirming whitening activity related with melanin content.

Effects of Combined Treatments of Lithium and Valproate on the Phosphorylation of ERK1/2 and Transcriptional Activity of ELK1 and C-FOS in PC12 Cells (리튬 및 발프로에이트 병용 처치가 PC12 세포에서 ERK1/2 인산화와 ELK1 및 C-FOS 전사활성에 미치는 영향)

  • Cha, Seung Keun;Kim, Se Hyun;Ha, Kyooseob;Shin, Soon Young;Kang, Ung Gu
    • Korean Journal of Biological Psychiatry
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    • v.20 no.4
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    • pp.159-165
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    • 2013
  • Objectives Mechanisms of clinical synergistic effects, induced by co-treatments of lithium and valproate, are unclear. Extracellular signal-regulated kinase (ERK) has been suggested to play important roles in mechanisms of the action of mood stabilizers. In this study, effects of co-treatments of lithium and valproate on the ERK1/2 signal pathway and its down-stream transcription factors, ELK1 and C-FOS, were investigated in vitro. Methods PC12 cells, human pheochromocytoma cells, were treated with lithium chloride (30 mM), valproate (1 mM) or lithium chloride + valproate. The phosphorylation of ERK1/2 was analyzed with immunoblot analysis. Transcriptional activities of ELK1 and C-FOS were analyzed with reporter gene assay. Results Single treatment of lithium and valproate increased the phosphorylation of ERK and transcriptional activities of ELK1 and C-FOS, respectively. Combined treatments of lithium and valproate induced more robust increase in the phosphorylation of ERK1/2 and transcriptional activities of ELK1 and C-FOS, compared to those in response to single treatment of lithium or valproate. Conclusions Co-treatments of lithium and valproate induced synergistic increase in the phosphorylation of ERK1/2 and transcriptional activities of its down-stream transcription factors, ELK1 and C-FOS, compared to effects of single treatment. The findings might suggest potentiating effects of lithium and valproate augmentation treatment strategy.

Eupatorium chinensis var. simplicifolium Root Extract Inhibits the Lipopolysaccharide-Induced Inflammatory Response in Raw 264.7 Macrophages by Inhibiting iNOS and COX-2 Expression (Raw 264.7 대식세포에서 등골나물 뿌리 추출물의 염증반응 조절 분자 iNOS와 COX-2 발현 억제 효과)

  • Lee, Jin-Ho;Kim, Dae-Hyun;Shin, Ji-Won;Park, Sae-Jin;Kim, Yoon-Suk;Shin, Yu-Su;Yu, Ji-Yeon;Kim, Tack-Joong
    • Journal of Life Science
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    • v.22 no.9
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    • pp.1137-1144
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    • 2012
  • Inflammation is a host defense mechanism that is activated in response to harmful substances or pathogens. However, an excessive inflammatory response is a problem in itself. Macrophages secrete inflammatory mediators such as nitric oxide (NO) or cytokines through various pathways such as the nuclear factor kappa B (NF-${\kappa}B$)-activated pathway after recognizing pathogen-like lipopolysaccharides (LPSs). In this study, anti-inflammatory effects of Eupatorium chinensis var. simplicifolium (EUC) extracts were investigated using LPS-stimulated RAW 264.7 macrophages. The EUC root extract significantly reduced NO production, inducible nitric oxide synthase (iNOS) expression, and cyclooxygenase-2 expression in a concentration-dependent manner. In addition, the EUC root extract reduced phosphorylation of mitogen-activated protein kinases and protein kinase B, which is upstream of NF-${\kappa}B$. The EUC root extract also reduced the degradation of inhibitory kappa B. These results indicate that EUC root extract exerts anti-inflammatory effects, which are mediated by inhibition of iNOS expression and the NF-${\kappa}B$ pathway.

Mechanisms of Suppression of Matrix Metalloproteinases in UVB-Irradiated HaCaT Keratinocytes of Colored Rice Varieties (UVB에 조사된 HaCaT Keratinocytes에서의 유색미에 의한 Matrix Metalloproteinases 발현억제 메커니즘)

  • Choi, Eun-Young;Lee, Jae-Bong;Kim, Do-Hoon;Kwon, Yong-Sham;Cheon, Jung-Yoon;Lee, Jin-Tae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.5
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    • pp.562-571
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    • 2017
  • In this study, we investigated the anti-oxidant activities [electron-donating ability (EDA), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging ability, and reactive oxygen species (ROS) inhibitory activity], anti-wrinkle activities [collagenase inhibitory activity, suppression and/or phosphorylation of matrix metalloproteinases (MMPs), and mitogen-activated protein (MAP) kinase activity], and mRNA expression levels using reverse transcription-polymerase chain reaction (RT-PCR) assay in ultraviolet (UV) B ray ($50mJ/cm^2$)-irradiated human keratinocyte HaCaT cells. Josaengheugchal, Sinneungheugchal (SE), Shintoheug rice, Heugjinju rice, and Heugseol (HE) among colored rice varieties were reported to have excellent antioxidant properties. In the EDA and ABTS radical scavenging assays, extracts of the five colored rice varieties had scavenging activities of 72% at concentrations higher $50{\mu}g/mL$. In the collagenase inhibition assay, ethanol extracts of the five colored rice varieties showed high inhibitory effects of about 60% at concentrations higher $25{\mu}g/mL$. In the ROS inhibition assay, ethanol extracts of HE and SE showed very excellent inhibition efficacies at all concentrations. We determined molecular biological mechanisms of MMPs (MMP-1, -3, -8, and -13) and mitogen-activated protein kinase (MAPK) with HE, and the results show that HE suppressed expression of MMPs and phosphorylation of MAPK and increased expression of pro-collagen type I in UVB-irradiated cells. It was also confirmed by RT-PCR that HE reduced expression of MMPs mRNA. Therefore, these results suggest that HE has anti-wrinkle and collagen production effects and may be used as a material in the development of functional food and cosmetic industries.

The Effect of the Use of Concept Mapping on Science Achievement and the Scientific Attitude in Ocean Units of Earth Science (해양단원 개념도 활용 수업이 과학성취도 및 태도에 미치는 효과)

  • Han, Jung-Hwa;Kim, Kwang-Hui;Park, Soo-Kyong
    • Journal of the Korean earth science society
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    • v.23 no.6
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    • pp.461-473
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    • 2002
  • Concept mapping is a device for representing the conceptual structure of a subject discipline in a two dimensional form which is analogous to a road map. In the teaching and learning of earth science, each concept depends on its relationships to many others for meaning. Using concept mapping in teaching helps teachers and students to be more aware of the key concepts and relationships among them. The purpose of this study is to investigate the effect of the use of concept mapping on science achievement and the scientific attitude in ocean units of earth science. The results of this study are as follows; first, the science achievement of a group of concept mapping teaching is significantly higher than that of the group of traditional teaching. Also, when the achievement levels are compared among different cognitive ability groups, the effect is more significant in mid or lower level student groups than in high level groups. The use of concept mapping is more effective when the concepts have a distinct concept hierarchy. Second, the scores of the test of ‘attitude toward scientific inquiry’ and ‘application of scientific attitude’ of the group of concept mapping teaching are significantly higher than those of the group of traditional teaching, whereas the scores of the test of ‘interest in science learning’ of concept mapping teaching is not different from those of group of traditional teaching. Third, the survey on the use of concept mapping shows a positive response across the tested groups. The use of concept mapping is more beneficial in fostering the comprehension of the topic. A concept map of student's own construction facilitates the assessment of learning, thus promising the usefulness of concept mapping as a means of evaluation. In regard to retention aspect, concept mapping is considered to be more effective in confirming and remembering the topic, while less effective in the aspects of activity and interest. In conclusion, the use of concept maps makes learning an active meaningful process and improves student's academic achievement and scientific attitude. If the concept mapping is more effectively as an active teaching strategy, more meaningful learning will be attained.