• 제목/요약/키워드: EROD activity

검색결과 76건 처리시간 0.03초

Reponses of the Hepatic Microsomal Cytochrome P450 Monooxygenase System in Rock Bream Oplegnathus fasciatus Exposed to Tributyltin (TBT)

  • Hwang, Un-Gi;Lee, Jung-Sik;Kang, Ju-Chan
    • Fisheries and Aquatic Sciences
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    • 제16권4호
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    • pp.261-265
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    • 2013
  • The study was conducted to investigate the responses of the hepatic microsomal cytochrome P450 monooxygenase system in the rock bream Oplegnathus fasciatus after chronic exposure to 0, 1, 2, 4, and $8{\mu}g/L$ tributyltin (TBT) concentrations for 4 weeks. Hepatic cytochrome 450 content and ethoxyresorufin O-deethylation (EROD) activity were found to significantly increase in fish treated with the higher concentration of TBT (${\geq}4{\mu}g/L$); however, no significant changes were observed in penthoxyresorufin O-deethylation (PROD) activity in all treated groups compared to the control group. These findings suggest that exposure to a low TBT concentration (${\geq}4{\mu}g/L$) has the potential to induce cytochrome 450 content and EROD enzyme activity in hepatic tissue in the rock bream.

ANTAGONISTIC EFFECTS OF INTERFERONS (INFs) AND SODIUM ORTHOVANADATE ON RESPONSES PRODUCED BY TCDD IN SEVERAL CULTURE SYSTEMS

  • Kim, Hwan-Mook
    • Toxicological Research
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    • 제7권2호
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    • pp.239-255
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    • 1991
  • Several types of IFNs were tested for their ability to suppress TCDD-inducible P-450 dependent mixed function oxidase (MFO) system in mouse primary hepatocytes. Mouse IFN-gamma (IFN-G) markedly suppressed EROD activity when added at the same time as TCDD (10 nM). The antagonism of EROD activity by IFN-G exhibited both a dose-(10-100 U/ml) and time-depedence. In contrast, mouse IFN-A/B was only moderately suppressive and only at high concentrations (500 U/ml). Rat IFN-G was even more selective than mouse, wherase human IFN-G had no activity.

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Inhibition of 7-Alkoxyresorufin O-Dealkylation Activities of Recombinant Human CYP1A1 and CYP1B1 by Resveratrol

  • Dong, Mi-Sook;Chang, Suk-Kyung;Kim, Hyun-Jung;F. Peter Guengerich;Park, Young-In
    • 한국환경성돌연변이발암원학회지
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    • 제22권3호
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    • pp.169-174
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    • 2002
  • Resveratrol is known to have potent cancer chemopreventive activity against tumorigenesis caused by 7,12-dimetylbenz[$\alpha$]anthracene(DMBA) which is known to be oxidized to reactive products by cytochrome P450 1B1 (CYP1B1). The effects of resveratrol on the activity of recombinant human P450 1 family enzymes, expressed in Escherichia coli membranes with human NADPH-P450 reductase, were determined by measuring alkoxyresorufin O-dealkylation activity, e.g., ethoxyresorufin O-deethylation (EROD) CYP1A1, methoxyresorufin O-demethylation (MROD), CYP1A2, benzyloxyresorufin-O-debenzylation (BROD), CTP1B1. Resveratrol inhibited CYP1B1 and CYP1A1 activities in a dose-dependent manner with $IC_{50}$/ values of 59 and 10$\mu$M for EROD activity and 1.8 and 30$\mu$M for BROD activity, respectively. Resveratrol had only weak inhibitory effect on CYP1A2 activity ($IC_{50}$/ values of 0.44 mM for EROD and >2 mM for MROD). Furthermore, resveratrol did not affect NADPH-P450 reductase activity significantly. Resveratrol inhibited the CYP1B1-dependent EROD activity with a $K_{i}$ of 28 $\mu$M in a non-competitive type manner. these results suggest that resveratrol-derived inhibited of CYP1B1 and CYP1A1 activities may contribute to the suppression of DMBA inducible tumorigenesis observed in extrahepatic tissues.s.

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노닐페놀을 주사한 조피볼락의 간장 cytochrome P450과 EROD의 반응 (Responses of Cytochrome P450 and EROD Activity in Rockfish (Sebastes schlegeli) Administered Intraperitoneal Injection of 4-nonylphenol)

  • 전중균;이지선;손영창;심원준;정지현;홍경표;김병기;한창희
    • 환경생물
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    • 제22권1호
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    • pp.171-176
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    • 2004
  • 비이온성 계면활성제로 많이 쓰이는 알킬페놀류의 하나인 노닐페놀(nonylphenol)이 해산어류에게 미치는 영향을 조사하려고 주요 양식어종인 조피볼락에게 복강주사로 10 및 25 mg $kg^{-1}$을 1회 투여하였다. 한편 용제인 DMSO만을 주사한 sham구를 설정하여 비교하였다. 주사 후 7일간 간중량지수(hepatosomatic index)의 변화를 조사하였고, 또한 간장 미크로좀 중 대표적인 약물대사효소인 cytochrome P450 (CYP) 농도와 ethoxyresorufin deethylase (EROD) 활성의 변화도 측정하였다. 그 결과, 고농도(25 mg $kg^{-1}$) 주사구에서는 HSl가 감소하였는데, 이것은 NP가 간장에 독성이 있다는 것을 보여준다. 한편, 약물대사효소 중 CYP 농도는 유도되었지만 EROD 활성은 저해되는 경향을 보였다. 이번에 주사한 농도는 우리나라 연근해 해수 중의 NP 농도에 비해 매우 높은 수준이지만, 본 실험에서는 NP가 조피볼락에 게간장 독성을 보였고, 간장의 약물대사효소에도 영향을 미친다는 것을 확인할 수 있었다.

Bioassays of Polycyclic Aromatic Hydrocarbons using CYP1A1-luciferase Reporter Gene Expression System in Human Breast Cancer MCF-7 Cells

  • Kim, Ja-Y.;Sheen, Yhun-Y.
    • 한국환경성돌연변이발암원학회지
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    • 제23권2호
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    • pp.45-50
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    • 2003
  • Biological activities of PAHs are not known although PAHs are considered as carcinogens. Recent industrial society has human widely exposed to PAHs (polynuclear aromatic hydrocarbons) that are comming from the incomplete combustion of organic material as wider spread environmental contaminants. Our laboratory have been studied the effect of PAHs in the human breast cancer MCF-7 cells. In this study, we examined the human breast cancer MCF-7 cells as a new system to evaluate bioactivity of PAHs. We have selected 13 PAHs to examine bioassay using CYP1A1-luciferase reporter gene expression system where CYP1A1 1.6 Kb 5flanking region DNA was cloned in front of luciferase reporter gene and this plasmid was transfected into MCF-7 cells transiently. This cells then used for the study to observe the effect of PAHs. We demonstrated that PAHs induced the CYP1A1 promoter, CYP1A1 mRNA and 7-ethoxyresolufin O-deethylase (EROD) activities in a concentration-dependant manner. None of PAHs that we have tested showed stronger stimulatory effect on CYP1 gene expression than TCDD. Benz(a)anthracene and benzo(b)fluoranthene were weak responders to CYP1A1 promoter activity stimulation, CYP1A1 mRNA and EROD induction in MCF-7 cells and these chemicals seemed to respond less either CYP1A1 mRNA or EROD than CYP1A1 promoter activity. Benzo(k)fluoranthene, chrysene, and dibenzo(a, h)anthracene showed strong response to CYP1A1 promoter activity stimulation, CYP1A1 mRNA increase and also EROD induction in MCF-7 cells. Results of dose response study suggested that two strong responding PAHs, such as benzo(k)fluoranthene and dibenzo(a, h)anthracene might be mediated through Aryl hydrocarbon receptors system in MCF-7 cells.

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Bioassays of Polycyclic Aromatic Hydrocarbons Using cyp1a1-Luciferase Reporter Gene Expression System in Mouse Liver Hepa 1 Cells

  • Min, Kyung-N.;Kim, Ja-Y.;Sheen, Yhun-Y.
    • 한국환경성돌연변이발암원학회지
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    • 제23권1호
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    • pp.30-34
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    • 2003
  • Recent industrial society has human widely exposed to PAHs (polynuclear aromatic hydrocarbons) that are comming from the incomplete combustion of organic material as wider spread environmental contaminants. Biological activities of PAHs are not known although PAHs are considered as carcinogens. Our laboratory have been studied the effect of PAHs in the mouse liver hepa 1 cells. In this study, we examined the mouse liver hepa-l cells as a new bioassay system to evaluate bioactivity of PAHs. We have selected 13 PAHs to examine bioassay using cyp1a1-luciferase reporter gene expression system where cyp1a1 1.6 Kb 5flanking region DNA was cloned in front of luciferase reporter gene and this plasmid was transfected into hepa 1 cells transiently. This cells then used for the study to observe the effect of PAHs. We demonstrated that PAHs induced the CYP1A1 promoter and 7-ethoxyresolufin O-deethylase (EROD) activities in a concentration-dependant manner. Some of PAHs showed stronger stimulatory effect on CYP1 gene expression than TCDD. Acenaphthene, anthracene, fluorine, naphthalene, pyrene, phenanthrene, carbazole were weak responders to cyp1a1 promoter activity stimulation and EROD induction in hepa 1 cells and these chemicals seemed to respond less to EROD than cyp1a1 promoter activity. Benz(a)anthracene, benzo(b)fluoranthene, benzo(k)fluoranthene, chrysene, and dibenzo(a,h)anthracene showed strong response to cyp1a1 promoter activity stimulation and also EROD induction in hepa 1cells. Results of dose response study suggested that four strong responding PAHs, such as benzo(a)anthracene benzo(k)fluoranthene, chrysene, and dibenzo(a, h)anthracene might be mediated through arylhydrocarbon receptor system in hepa1 cells.

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Tin-free 방오제인 Sea-Nine 211을 주사한 북방대합에서 alkoxyresorufin 탈알킬화효소와 글루타치온 포합효소 활성의 변화 (Responses of Alkoxyresorufin Dealkylases and Glutathione S-transferase Activities of Surf Clam, Pseudocardium sachalinensis, Injected with Sea-Nine 211 Antifoulant)

  • 이지선;전영하;심원준;전중균
    • 환경생물
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    • 제26권2호
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    • pp.109-114
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    • 2008
  • 방오도료로 많이 쓰이던 유기주석화합물은 일반생물에게 미치는 독성이 매우 강하고 또한 내분비계 장애물질임이 밝혀지면서 이를 대체할 화합물들이 개발되고 있다. 그 가운데 본 연구에서는 Sea-Nine 211(4,5-dichlore-2-n-octyl-3(2H) isothiazolone)을 사용하여 이 화합물이 해양생물 특히 저서생물인 패류에게 얼마나 영향을 미치는지를 살펴보았다. 이를 위해 강원도 북부 해역에 주로 서식하는 북방대합(Pseudocardium sachalinensis)에게 Sea-Nine 211을 강제적으로 주사하여 alkoxyresorufin 탈알킬화효소인 EROD와 MROD의 활성 변화를 4일째까지 조사하였고, 비교를 위해서 tributyltin chloride (TBTC)도 사용하였다. 그 결과, CYP1A1의 지표효소인 EROD활성은 Sea-Nine 211 및 TBTC주사구 모두 sham구와 차이가 없었지만, CYP1A2의 지표효소인 MROD활성은 Sea-Nine 211 주사구가 sham구에 비해 유의적으로 높았으므로 Sea-Nine 211에 의해 유도된 것이라 여겨진다. 이것은 북방대합에도 CYP1A2가 존재할 수 있음을 보여준다.

Establishment of CALUX Bioassay for Dioxin Determination

  • Joung, Ki-Eun;Chung, Young-Hee;Sheen, Yhun-Yhong
    • 한국환경성돌연변이발암원학회지
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    • 제24권3호
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    • pp.137-142
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    • 2004
  • Dioxin-like compounds are ubiquitous environmental polltants that could be accumulated in biological system and toxic to human and wildlife. Given this issue, it is important to develop a reliable dioxin detection methods for a rational risk assesment of dioxin-like compounds. In this study, we tried to set up and validate a sensitive, reliable risk assessment of dioxin-like compounds. In this study, we tried to set up and validate a sensitive, reliable and rapid bioassay model, CALUX bioassay as a screening tool for routine measurement of dioxin-like conpounds in environmental matrices. For the valisation of CALUX bioassay, firstly, we performed dose-response assay for 2,3,7,8-TCDD, most potent dioxin-like compound, using two different methods CALUX and EROD assay. Induction of luciferase activity and CYPIA catalyzed EROD activity were dose-dependently induced by 2,3,7,8-TCDD, with initial induction at 0.1 pM and maximal induction at 1 nM. In order t determine whether the CALUX bioassay could predict the effects of dioxin-like compounds, 2,3,7,8-TCDD dose-response from CALUX was compared with that from EROD assay. The correlation coefficient ($r^2$) was found to be 0.89, indicating a good correlation between two different methods and the possibility of CALUX bioassay as a useful dioxin detecting method.

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Morin이 benzo(k)fluoranthene에 의한 유방암 세포 MCF-7의 CYP1A1 유전자 발현 조절에 미치는 영향 (Effects of Morin on benzo(k)fluoranthene Regulated CYP1A1 Gene Expression in MCF-7 Human Breast Cancer Cells)

  • 양소연;김여운;신윤용
    • 한국환경성돌연변이발암원학회지
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    • 제24권4호
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    • pp.189-197
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    • 2004
  • We investigated the effect of dietaty flavonoid, such as CYP1A1 promoter activity, 7-ethoxyresorufin-O-deethylase(EROD) activity and CYP1A1 mRNA expression induced by benzo(k)fluoranthene(B(k)F) in MCF-7 cells. Based on the three criteria of frequency of occurrence in the environment, toxicity and potential exposure to humans, B(k)F is one of the top-listed PAHs. We found that B(k)F significantly up-regulates the level of CYP1A1 promoter activity, EROD and CYP1A1 mRNA. When cells were treated with morin alonem, it was not changed that EROD and CYP1A1 mRNA, compared to that of control. However, morin inhibited the B(k)-induced CYP1A1 prompter activity and mRNA level at high concentration. But morin exhibited stimulatory effects B(k)F-induced CYP1A1 promoter activity and mRNA level at low concentration. Overall, results from these studies demonstrate morin might interfere the action of B(k) with AhR system to stimulate CYP1A1 gene expression. CYP1A1 is known to be inducible by xenobiotic compouds such as polyciclic aromatic hydrocarbons(PAHs) and 2,3,7,8-tetrachloro-dibenzo-p-dioxin(TCDD). These chemicals have been identified worldwide and can have a significant impact on the human health and well being of human and wildlife. Given these issues, the detection and quantification of these chemicals in biological, environmental and food samples is important.

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