• 제목/요약/키워드: cytosol

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p38 MAPK을 이용한 항염증 효능 규명 프로토타입 개발 (Development of Prototype for Screening Anti-Inflammation Effects concerning p38 MAPK Signal Pathway)

  • 김철;예상준;남기엽;김상균;장현철;김진현;김영은;송미영
    • 한국한의학연구원논문집
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    • 제17권3호
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    • pp.77-85
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    • 2011
  • Objectives : The purpose of this study was to develop a simulator which can analyze the anti-inflammatory effects of medical herbs based on e-cell concerning p38 MAPK signal pathway. Methods : We collected data concerning medical herbs with anti-inflammatory effects and the active compounds to provide as a fundamental databse and to validate the newly developed algorithm. At this time, we used the target database as pubmed and gathered the data by data mining tool, pathway studio. Also we have developed the web-based search system for confirming database related to anti-inflammation. We researched the mechanism of actions of proteins in p38 MAPK signal pathway when active compound has been inserted into the network. We reduced total network into TAK-MKK3-p38 and made the two types of mathematical model about active compounds' interaction. Results & Conclusion : We constructed the database which have 69 cases of medical herbs, 71 cases of active compounds, about 8,000 cases of URL(Uniform Resource Locator) related to papers and reports. We designed the ordinary differential equations for response of TAK, MKK3, p38 in e-cell's cytosol and nucleus. We used this formular as measure whether an active compound of medicinal plants which is inputted by an user would have an anti-inflammation effects. We developed the visualization program which could show the change of concentration over time.

Effects of Selenizing Codonopsis pilosula Polysaccharide on Macrophage Modulatory Activities

  • Qin, Tao;Ren, Zhe;Lin, Dandan;Song, Yulong;Li, Jian;Ma, Yufang;Hou, Xuehan;Huang, Yifan
    • Journal of Microbiology and Biotechnology
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    • 제26권8호
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    • pp.1358-1366
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    • 2016
  • The purpose of the present study was to investigate the immune-enhancing activity of selenizing Codonopsis pilosula polysaccharide (sCPPS5) in nonspecific immune response. In in vitro experiment, the results showed that sCPPS5 could promote the phagocytic uptake, NO production, and TNF-α and IL-6 secretion of RAW264.7 cells. sCPPS5 could also strongly increase the IκB-α degradation in the cytosol and the translocation of NF-κB p65 subunit into the nucleus of RAW264.7 cells. In the vivo experiment, sCPPS5 at medium doses could significantly improve the phagocytic index of peritoneal macrophages and induce the secretion of TNF-α and IL-6. Moreover, the effect of sCPPS5 was significantly better than Codonopsis pilosula polysaccharide (CPPS). These results indicated that selenylation modification could significantly enhance the immune-enhancing activity of CPPS in the nonspecific immune response.

미역의 급여 수준이 타고난 면역반응이 활성화한 육계병아리의 혈액 항산화 균형에 미치는 영향 (Effect of Dietary Brown Seaweed Levels on the Antioxidant System in Broiler Chicks Activated Innate Immune Response)

  • 이혜정;박인경;임진택;최도열;최준영;최종배;이홍구;최윤재;고태송
    • Journal of Animal Science and Technology
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    • 제47권1호
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    • pp.29-38
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    • 2005
  • Effect of dietary brown seaweed(Undaria pinnatifida) levels on the anti-oxidant enzyme system was evaluated in blood of broiler chicks activated innate immune response. Day-old broiler chicks were fed diets containing 0.0(basal), 1.0, 2.0 and 4.0 % of brown seaweed for 4 weeks. The innate immune response was activated by injecting Salmonella typhymurium lipopolysaccharide(LPS) i.p. at 8, 10 and 12 day of age. The activation of innate immune response enhanced(p< 0.01) and the brown seaweed 1.0 and 2.0 % diets reduced(P< 0.05) the superoxide dismutase(SOD) activity in erythrocyte cytosol significantly. The activation of innate immune response elevated significantly the levels of peroxide and the activity of peroxidase in plasma, while the levels of dietary brown seaweed resulted in a significant linear increase in peroxidase activity in plasma of normal bird. Experience of the innate immune response in 4 week-old chicks reduced linearly the plasma level of peroxide with the level of brown seaweed and enhanced the plasma peroxidase activity. Also, the plasma of normal birds fed the brown seaweed showed higher level of peroxide and lower activity of peroxidase. The results indicated that dietary brown seaweed affected SOD and peroxidase activities in blood of broiler chicks during activation of innate immune response.

Involvement of Caspases and Bcl-2 Family in Nitric Oxide-Induced Apoptosis of Rat PC12 Cells

  • Jeong, Yeon-Jin;Jung, Ji-Yeon;Lee, Jin-Ha;Cho, Jin-Hyoung;Lee, Guem-Sug;Kim, Sun-Hun;Kim, Won-Jae
    • The Korean Journal of Physiology and Pharmacology
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    • 제10권6호
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    • pp.329-335
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    • 2006
  • This study was aimed to investigate the nitric oxide (NO)-induced cytotoxic mechanism in PC12 cells. Sodium nitroprusside (SNP), an NO donor, decreased the viability of PC12 cells in dose-and time-dependent manners. SNP enhanced the production of reactive oxygen species (ROS), and gave rise to apoptotic morphological changes including cell shrinkage, chromatin condensation, and DNA fragmentation. Expression of Bax was not affected, whereas Bcl-2 was downregulated in SNP-treated PC12 cells. SNP augmented the release of cytochrome c from mitochondria into cytosol and enhanced caspase -8, -9, and -3 activities. SNP upregulated both Fas and Fas-L, which are known to be components of death receptor assembly. These results suggest that NO induces apoptosis of PC12 cells through both mitochondria-and death receptor-mediated pathways mediated by ROS and Bcl-2 family.

길경이 인체 폐암세포에 미치는 영향에 대한 실험적 연구 (Apoptotic Cell Death of Human Lung Carcinoma A549 Cells by an Aqueous Extract from the Roots of Platycodon grandiflorum)

  • 이성열;김원일;박동일
    • 동의생리병리학회지
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    • 제17권4호
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    • pp.1019-1030
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    • 2003
  • Platycodi Radix, the root of Platycodon grandiflorum, commonly known as Doraji, is used as a traditional oriental medicine. Extracts from the roots of P. grandiflorum have been reported to have wide ranging health benefits. In the present study, we investigated the effects of an aqueous extract from the roots of P. grandiflorum (AEPG) on the growth of human lung carcinoma A549 cells. Results obtained are as fellow; AEPG treatment resulted in the inhibition of the cell viability of A549 cells in a concentration-dependent manner. Upon treatment with AEPG, A549 cells developed many of the hallmark features of apoptosis, including condensation of chromatin. Flow cytometry analysis confirmed that AEPG increased populations of apoptotic-sub G1 phase. Western blot and RT-PCR analyses indicated that the expressions of Bcl-2 was down-regulated but Bax was up-regulated in AEPG-treated A549 cells. AEPG-induced apoptotis of A549 cells was associated with rroteolytic cleavage and activation of caspase-3, release of cytochrome c from mitochondria into cytosol and down-regulation of Akt and phospho-Akt proteins in a dose-dependent manner. Induction of apoptosis by AEPG treatment was associated with inhibition and/or degradation of apoptotic target proteins such as poly(ADP-ribose) polymerase, β-catenin and phospholipase C-γ 1. AEPG treatment inhibited the levels of cyclooxygenases protein of A549 cells, which was associated with the inhibition of prostaglandin E2 accumulation in a concentration-dependent fashion. Taken together, these findings suggest that P. grandiflorum has strong potential for development as an agent for prevention against human lung cancer.

폐흡충 성충 Cu, Sn-Superoxide Dismutase의 정제 및 생화학적 특성 (Purification and Characterizatlon of a Cu, Zn-Superoxide Dismutase from Adult Paragonimus westermani)

  • 정영배;송철용
    • Parasites, Hosts and Diseases
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    • 제29권3호
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    • pp.259-266
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    • 1991
  • 폐흡충 성충의 인산완충액 추출액을 원심분리하여 만든 세포질 현탁액을 조효소(조효소)로 사용하여 Xanthine- zanthine oxidase system으로 superoxide digmutase 환성을 측정한 결과 비활성도(비활성도: specific activity)는 4.3 units/mg이었다. 이 효소의 활성도를 30∼85% ammonium sulfate 침전 및 DEAE-Trisacryl M anion exchange chromatography와 Sephadex G-100 column chromatography를 통과시키면서 측정하는 방법으로 superoxide dismutase를 정제하고, 정제한 효소의 생화학적 특성을 관찰하여 다음과 같은 결과를 얻었다. 1. 정제과정을 통해 세포질 내 superoxide dismutase를 150배 정제하였다. Sephadex G-100 gel filtration에 의해 계산한 이 효소의 분자량은 34kDa이었고 환원성 SDS-PAGE상 subunit가 17 kDa이었다. 따라서 이 효소는 subunit 두개로 구성된 중항체로 판단하였다. 3. 이 효소는 1 mM 이상의 cyanide 농도에서는 효소 황성이 100% 억제되었고, 5mM 농도의 azide에서는 11.1%, 10mM azide에서는 22.2% 그 환성이 각각 억제되었다. 3. 이 효소는 완충액의 pH가 10.0일 때 효소 황성이 5.4배 증가되었다. 이상의 결과로 폐흡충의 세포질 내에 존재하는 superoxide dismutase는 구리와 아연을 함유한 superoxide dismutase라고 판단할 수 있었다.

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Hsp70 분자 샤페론과 조절인자 (Family of Hsp70 Molecular Chaperones and Their Regulators)

  • 정경태
    • 생명과학회지
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    • 제17권12호
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    • pp.1760-1765
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    • 2007
  • 생명체 내에서 일어나는 거의 모든 반응은 단백질이 촉진하거나 수행한다. 단백질은 세포질과 소포체에서 합성될 때 엄격하게 조절된다. 그러나, 새로이 합성된 모든 단백질이 살아남아서 생명을 유지시키는 기능에 관여하게 되지는 않는다. 가장 알맞은 생리학적 in vitro 실험 조건에서 새로이 합성된 단백질의 약 3분의 1 정도는 합성되자마자 proteasome에 의해 빠르게 분해된다고 보고되었다. 또한, 단백질은 합성이 성공적으로 이루어진 이후에는 3차원 구조를 갖기 위해 접힘(folding)이 이루어져야 하고, subunit들은 assembly 과정을 거쳐야 비로소 성숙된 단백질로서 기능을 하게 된다. 어떤 단백질군은 자연적으로 접힘이 일어나는 반면 어떤 단백질군은 분자 샤페론(molecular chaperones)과 folding enzymes의 도움을 받아야만 접힘이 일어난다. 분자 샤페론은 세포 전역에 분포하고 있으며, 세균에서부터 고등 동식물에 이르기까지 모든 생명체에 존재한다. 이들 중 Hsp70군은 많이 연구된 분자 샤페론으로서 지난 10여년 동안 조절인자들이 새로이 발견되어 작용 mechanism이 보다 자세히 밝혀졌다. 본 총설에서 Hsp70군과 그 조절인자들에 대한 전반적인 서술을 하였으며, 이들의 기능이 분자 샤페론 기능 외에 생체 내에서 중요한 기능들이 새롭게 밝혀지고 있어 이들의 작용 mechanism을 조명함으로 이해를 돕고자 한다.

Artemia franciscana cyst의 세포내 산성 Protease (Characterization of Cytosol Acid Protease from Dormant Cyst of Artemia franciscana)

  • 최희선;최경희;류태형
    • 생명과학회지
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    • 제7권3호
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    • pp.228-233
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    • 1997
  • Artemia franciscana의 cyst에서 protease를 정제하가 위해 세포를 파쇄하여 원심 분리한 후 상등액을 회수하였다. 그상등액에서 40-60% 황산암모늄을 첨가하여 침전시켜서 염을 제가한 후, Sphadex G-200 및 DEAE-Sephadex A-50 column chromatography를 하여 정제한 후 본 효소의 특성을 조사한바 그 특성은 아래와 같았다. 정제 과정을 거치는 동안, 비활성은 13.64배 증가하였고, 수율은 7%였다. 그리고 이효소는 pH 6에서는 안정하였으나, pH 8이상에서는 대부분의 활성이 실활 되었고, 최적 pH는 3.0이었다. 따라서 이 효소는 전형적인 산성 pro-tease로 판정되었다. 본 효소는 60$^{\circ}$C이상의 온도에서 대부분 실활 되었고, 최적 활성 반응 온도는 35$^{\circ}$C였다. 금속 이온의 영향을 알아본 결과, $Cu^{++}$, Z$Zn^{++}$, $Fe^{++}$가 효소의 활성을 저해하였고, 중금속 chelator인 EDTA는 반대로 효소 활성을 증가시켰다. 단백질분해효소에 특이적인저해제를 첨가하여 실험한 결과 thiol protease ingibitor인 antipain, chymostatin, leupeptin, E-64, iodoacetate등에 효과적으로 효소활성이 저해받으므로 본 효소는 acid thiol protease라 판정했다.

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Effects of Copper, Zinc and Cadmium on the Recovery Pattern of Aryl Sulfotransferase IV Activity in Rats fed 2-Acetylaminofluorene Diet

  • Chung Keun Hee;Ringel David P.;Shin Kyung Ok
    • Nutritional Sciences
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    • 제9권1호
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    • pp.29-34
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    • 2006
  • Purified rat liver aryl sulfotransferase IV (AST IV) was found to be inhibited in vitro by zinc, copper, cadmium and terbium. Among these four elements, zinc, copper and cadmium were all strongly inhibitory to the AST IV activity at very low concentrations (2.5 $\mu$M to 0.025 $\mu$M). In rat liver cytosol, zinc, copper and cadmium at 25 $\mu$M to 0.025 $\mu$M also decreased the AST IV activity to $50\%$ of the controls. In order to assess the possible effects of these metals on the AST IV activity recovery pattern in vivo, studies on the relationship between these minerals and dietary 2-acetylaminofluorene were conducted. Total of forty rats were fed one of five diets for 6 weeks: diet 1, Control diet plus 2-acetlyaminofluorene ($0.05\%$); diet 2, zinc-deficient diet plus 2-acetlyaminofluorene; diet 3, zinc-supplement diet plus 2-acetylaminofluorene; diet 4, copper-supplement diet plus 2-acetylaminofluorene; diet 5, cadmium-supplement diet plus 2-acetylaminofluorene. Half of the rats from each diet were changed to individual diet after 3 weeks of 2-acetylaminofluorene feeding. Placement of rats on the control diet following one cycle of 2-acetylaminofluorene feeding of 3 weeks without 2-acetylaminofluorene resulted in nearly full recovery of AST IV activity within 3 or 4 weeks. However, the rats fed diets that supplemented with zinc, copper or cadmium without 2-acetylaminofluorene showed a new pattern of lowered AST IV activity as early as the first cycle. Also, lowering in cytosolic AST IV contents was appeared in the livers from the rats, following one cycle of 2-acetylaminofluorene feeding of 3 weeks, fed one of the diets that supplemented with copper, cadmium or zinc without 2-acetylaminofluorene for ensuing 3 weeks.

Effect of Biochanin A on the Aryl Hydrocarbon Receptor and Cytochrome P450 1A1 in MCF-7 Human Breast Carcinoma Cells

  • Han, Eun-Hee;Kim, Ji-Young;Jeong, Hye-Gwang
    • Archives of Pharmacal Research
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    • 제29권7호
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    • pp.570-576
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    • 2006
  • Phytoestrogen biochanin A is an isoflavone derivative isolated from red clover Trifolium pratense with anticarcinogenic properties. This study examined the action of biochanin A with the carcinogen activation pathway that is mediated by the aryl hydrocarbon receptor (AhR) in MCF-7 breast carcinoma cells. Treating the cells with biochanin A alone caused the accumulation of CYP1A1 mRNA and an increase in CYP1A1-specific 7-ethoxyresorufin O-deethylase (EROD) activity in a dose dependent manner. A concomitant treatment with 7,12-dimethylbenz[a]anthracene (DMBA) and biochanin A markedly reduced the DMBA-inducible EROD activity and CYP1A1 mRNA level. In addition, the biochanin A treatment alone activated the DNA-binding capacity of the AhR for the dioxin-response element (DRE) of CYP1A1, as measured by the electrophoretic-mobility shift assay (EMSA). EMSA revealed that biochanin A reduced the level of the DMBA-inducible AhR-DRE binding complex. Furthermore, biochanin A competed with the prototypical AhR ligand, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), for binding to the AhR in an isolated rat cytosol. The biochanin A competitively inhibited the metabolic activation of DMBA, as measured by the formation of the DMBA-DNA adducts. These results suggest that biochanin A may thus be a natural ligand to bind on AhR. Therefore, biochanin A may be due to act an antagonist/agonist of the AhR pathway.