• 환경생물
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• 제22권1호
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• pp.177-183
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• 2004

유기주석화합물인 tributyltin chloride (TBTC), tributyltin oxide (TBTO)와 triphenyltin chloride (TPTC)를 넙치 간장으로 만든 미크로좀에 in vitro적으로 노출시켜서 이들 화합물의 대사에 관여하는 mixed function oxidase (MFO) 중 cytochrome P450 (CYP) 농도와 7-ethoxyresorufin deethylase (EROD) 활성의 변화를 조사하였으며, 또한 in vivo 실험에서는 TPTC를 넙치에게 복강주사(7.5 mg $kg^{-1}$ BW)하여 간장의 MFO (CYP농도, NADPH cytochrome c 환원효소 활성, NADH chtochrome b5 환원 효소 활성, EROD 활성) 반응을 경시적으로 조사하였다. 그 결과, in vitro에서는 TBTC, TBTO 및 TPTC가 모두 CYP 농도와 EROD 활성을 저해하였으며, 저해력은 TPTC가 가장 컸고 이어서 TBTO, TBTC의 순이었다. 유기주석화합물의 노출농도와 노출시간과 비례하면서 저해정도가 커졌으며, 특히 EROD활성의 저해는 노출농도에 크게 의존적이었다. 그리고 in vivo실험에서도 유기주석 화합물은 CYP농도, NADPH cytochrome c 환원효소 활성, NADH cytochrome b5 환원효소 활성, EROD 활성을 억제하였다. EROD 활성은 오염물질에 의한 반응이 민감하고 재현성도 있어 바람직한 측정지표로 이용될 수가 있을 것이다.

• Archives of Pharmacal Research
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• 제26권12호
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• pp.1079-1086
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• 2003

The aim of this study was to investigate the effect of trauma on cytochrome P450 (CYP) gene expression and to determine the role of Kupffer cells in trauma-induced alteration of CYP isozymes. Rats underwent closed femur fracture (FFx) with associated soft-tissue injury under anesthesia. To deplete Kupffer cells in vivo, gadolinium chloride ($GdCl_3$) was intravenously injected at 7.5 mg/kg body wt., 1 and 2 days prior to FFx surgery. At 72 h of FFx, liver tissues were isolated to determine the mRNA and protein expression of CYP isozymes and NADPH-P450 reductase by reverse transcription-polymerase chain reaction and Western immunoblotting, respectively. In addition, the mRNA levels of tumor necrosis factor alpha (TNF-$\alpha$), inducible nitric oxide synthase (iNOS) and heme oxygenase-1 (HO-1) were evaluated. FFx increased the mRNA level of CYP1A1; an increase that was not prevented by $GdCl_3$. There were no significant differences in the mRNA expression of CYP1A2, 2B1 and 2E1 among any of the experimental groups. The protein levels of CYP2B1 and 2E1 were significantly decreased by FFx; a decrease that was not prevented by $GdCl_3$ treatment. The gene expression of NADPH-P450 reductase was unchanged by FFx. FFx significantly increased the expression of TNF-$\alpha$ mRNA; an increase that was attenuated by $GdCl_3$. The mRNA expression of HO-1 was increased by FFx, but not by $GdCl_3$ . Our findings suggest that FFx differentially regulates the expression of CYP isozyme through Kupffer cell-independent mechanisms.

• 약학회지
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• 제23권2호
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• pp.111-118
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• 1979

It was previously reported that cytochrome P$_{450}$ content in liver was increased when Capsicum acetone extract was given chronically to rats. The present study is aimed to investigate the effect of capsaicin, a principal component of red pepper, on the drug metabolizing enzymes in rat liver. Capsaicin (5mg/kg) was given intraperitoneally once a day for seven days and zoxazolamine paralysis time and hexobarbital sleeping time were determined 24 hrs after the last dose of capsaicin. Plasma hexobarbital concentration was also determined five and 15 min after hexobarbital administration to rats. Zoxazolamine paralysis time and hexobarbital sleeping time were shortened by 31.6% and 37.1%, respectively, compared with control group. Plasma hexobarbital concentration was lowered by 26.2% after five min and by 35.2% after 15 min, respectively, compared with control group. However, administration of single dose of capsaicin did not affect the zoxazolamine paralysis time and hexobarbital sleeping time. Microsomal cytochrome P$_{450}$ content and NADPH-cytochrome C reductase activity were increased by 14.6% and 11.6%, respectively in the rats pretreated with capsaicin for seven days, while cytochrome b$_{5}$ content was not changed. These results suggest that treatment with capsaicin for seven days may induce the drug metabolizing enzyme in rat liver.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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• pp.37-38
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• 2001

The cytochrome P450 (P450) 1 family (1A1, 1A2, 1B1) is involved in the activation of many pro-carcinogens. Previously we characterized a number of synthetic bi- and polycyclic hydrocarbon acetylenes as selective-mechanism-based inhibitors of recombinent P450s 1A1, 1A2, 1B1 (Shimada et al., Chem. Res. Toxicol., 11, 1048-1056, 1998). We reported that the drug oltipraz is a mechanism-based indicator of P450 1A2 (Lagouet et al. Chem. Res. Toxicol., 13, 245-252, 2000).(omitted)

• 한국발생생물학회지:발생과생식
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• 제11권3호
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• pp.195-203
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• 2007

자웅이체형 어류의 성결정 및 성분화는 일반적으로 각 개체의 유전형을 따른다. 자연환경에서는 자신이 가진 유전정보의 조절에 따라 성분화 시기에 아로마테이즈 유전자의 발현이 증가하거나 감소하고, 그 결과 스테로이드 호르몬들의 조성이 결정되어 각기 다른 방향으로 성분화하는 것으로 알려져 있다. 본 연구에서는 해산 태생 어류인 조피볼락(Sebasts schlegeli)의 ovarian type aromatase (P450aromA)와 brain type aromatase (P450aromB)의 유전자를 부분적으로 클로닝하여, 이들의 염기서열을 밝혔으며, 각 유전자에 대한 primer를 제작한 후 성분화 기간 중 이 유전자들의 발현을 조사하였다. 조피볼락 아로마테이즈 유전자들은 조사를 시작한 출산 후 35일째에 여러 개체의 머리와 몸에서 각각 발현되었으나, 52일째에는 아로마테이즈 유전자들의 발현 개체수가 현저히 감소하였다. 그리고 출산 59일째에는 발현 개체 수가 다시 증가하였다. P450aromA과 P450aromB의 발현 양상은 전반적으로 유사하였으나, 45일째에는 P450aromB의 발현 개체수가 P450aromA 발현 개체 수보다 훨씬 많았다. 조직학적 분석을 통해 제시된 이 종의 성분화 시기는 지금까지 출산 $50{\sim}65$일 전후로 알려져 왔으나, 본 연구에서 나타난 아로마테이즈 유전자들의 발현 결과를 볼 때, 이 종의 실질적인 성분화는 알려진 것보다 최소한 $1{\sim}2$주 정도 빨리 진행될 가능성이 있다. 본 연구의 결과는 또한 태생어류의 성분화도 난생어류의 성분화에서 보고된 것과 마찬가지로 아로마테이즈의 작용과 밀접하게 연관되어 있다는 사실을 시사한다.

• 대한수의학회지
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• 제38권1호
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• pp.17-28
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• 1998

Drug-metabolizing activities of Korean native cattle and swine were investigated from viewpoints of the cytochrome P-450's level, their dependent mixed function oxidase activities, the reactive oxygen species formation and cytosolic enzyme acitivities from each liver homogenates. Level of cytochrome P-450 in the liver microsome of Korean native cattle was $0.28{\pm}0.05nmole/mg$ and that in pigs $0.35{\pm}0.03nmole/mg$. Level of cytochrome $b_5$ of Korean native cattle was $0.24{\pm}0.06nmole/mg$, and that of pigs $0.2{\pm}0.05nmole/mg$, showing no difference between two species. NADPH P-450 reductase were higher in Korean native cattle ($58.3{\pm}5.3nmole/mg/min$) than in pigs ($29.9{\pm}3.8nmole/mg/min$)(p<0.01). The activities of cytochrome P-450 dependent monooxygenases such as ethoxyresorufin O-deethylase (cattle, $96.5{\pm}12.5nmole/mg/min$ ; pigs, $13.6{\pm}2.1nmole/mg/min$), N-benzphetamine N-demethylase (cattle, $5.23{\pm}0.82nmole/mg/min$ ; pigs, $0.76{\pm}0.3nmole/mg/min$) and aniline hydroxylase (cattle, $0.95{\pm}0.1nmole/mg/min$ ; pigs, $0.33{\pm}0.08nmole/mg/min$) were much higher in Korean native cattle than in swine(p<0.01). However, the activity of testosterone $7{\alpha}$-hydroxylase was higher in swine ($90.4{\pm}1.2nmole/mg/min$) than cattle (cattle, $32.8{\pm}1.2nmole/mg/min$). Interestingly, testosterone $16{\alpha}$-hydroxylase, a marker enzyme for P-450 IIA was not detected in both animal species. These results suggest that Korean native cattle and pigs have high contents of P-450 IA1 and P-450 IIIA. Total sulfhydryl compound (cattle, $10.3{\pm}1.1nmole/mg$ ; Pigs, $14.5{\pm}1.8nmole/mg$) and glutathione related enzymes except glutathione reductase (cattle, $38.1{\pm}7.9nmole/mg/min$; swine, $22{\pm}3.6nmole/mg/min$) showed higher levels in swine than in Korean native cattle. Superoxide dismutase (cattle, $7.64{\pm}0.84nmole/mg/min$ ; pigs, $4.47{\pm}0.94nmole/mg/min$) and catalase (cattle, $30.4{\pm}3.7nmole/mg/min$ ; pigs, $17.2{\pm}1.8nmole/mg/min$) were remarkably higher in Korean native cattle than in swine (p<0.05).

• 환경생물
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• 제21권1호
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• pp.18-25
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• 2003

본 연구는 내분비교란물질로 알려진 TBTC가 해양생물에게 미치는 영향을 조사하는 연구의 일환으로, 이 화합물에 노출시킨 어류의 간장 MFO 효소계의 반응을 in vitro로 조사하였다. 대상 어류는 개복치(Mola mola), 숭어(Mugil cephalus), 강도다리(Ptatichthys stellatus), 청어(Clupeu pallasii), 붕장어(Astroconger myriaster), 조피볼락 (Sebastes schlegeli), 참돔(Pugrus major), 넙치(Paralichthys olivaceus)이며, 이들의 간장 미크로좀을 2mM의 TBTC와 in vitro (3$0^{\circ}C$, 20분)로 배양하고 cytoch-rome P45O (CYP), cytochrome b5, NAD(P)H -cytochro-me c 환원효소를 비롯한 탈알킬화 효소들(EROD, PROD, MROD, ECOD)의 변화를 조사하였다. TBTC는 환원효소의 측정 시에는 DMSO에, 그리고 그 밖의 효소 측정시에는 메탄올에 녹여 2% 농도로. 첨가하였다. 어류의 간장 CYP 함량은 TBTC와 배양 후 대부분(6 어류/8어류)에서 10% 이하로 크게 저해되었으나, cyto-chrome b5함량은 변함이 없었다. 하지만 NAD(P)H 의존성 환원효소의 반응은 어류에 따라 달랐고, CYP 외에 두 환원효소도 모두 영향을 받아 저해되는 타입 1(개복치, 조피볼락, 청어, 강도다리, 참돔), CYP 외에 NADH 의존성 환원효소만 저해되는 타입 2(붕장어, 숭어)및 CYP 저해되지만 두 환원효소는 영향을 받지 않거나 오히려 유도되는 타입 3 (넙치)으로 구분되었다. 그리고 대부분(7/8)의 어류에서는 NADH 의존성 환원효소가 NADPH 의존성 환원효소에 비해 더욱 저해되는 경향을 보였다. TBTC는 어류의 탈알킬화 효소에도 영향을 미쳤고, 어류별 EROD활성의 저해는 개복치, 참돔, 붕장어, 조피볼락(잔존율 1~7%)>숭어, 청어 (14~30%)>넙치, 강도다리(56~65%)의 순이었으며, ECOD 활성의 저해도 개복치, 참돔, 붕장어 (36~38%)>조피볼락(63%))숭어, 청어, 넙치, 강도다리(90%)의 순으로 비슷한 경향이었다. 한편, 넙치와 강도다리에서는 PROD가 MROD보다 더욱 심하게 저해되었다. 이처럼 어류 간장의 약물대사 효소계는 TBTC에 의한 저해 정도가 어류에 따라 심한 차이를 보였다.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.312.1-312.1
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• 2002

Previous studies have demonstrated that 2,3',4,5' -tetramethoxystilbene (TMS) and 3,3',4',5,5'-pentamethoxystilbene (PMS) showed selective inhibition of human cytochrome P450 1 Bl and 1A1 in vitro., respectively, In the present study, the effects of synthetic stilbene analogs on the expression of cytochrome P450 1Al or lBl were investigated in human tumor cell lines such as HepG2, MCF-7 and MCF-l0A, TCDD caused a dramatic increase in the amount of P450 1A1 or 1B1 proteins and mRNA levels. (omitted)

• 한국독성학회:학술대회논문집
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• 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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• pp.188-188
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• 2001

Human cytochrome B5 (CYB5) was coexpressed with cytochrome P450 1A2 (CYP1A2), NADPH-CYP450 reductase (CYPR) and Ν-acetyltransferase 2 (NAT2) in Chinese hamster ovary (CHO) cells. The expression of four proteins was determined by Western blot analyses. The introduction of cDNAs to CHO cells were transduced via retroviral vectors. The cytotoxicity assay of 2-aminoanthracene (2-AA) and aflatoxin B$_1$were approximately 4-fold more sensitive than CYB5 free cells.(omitted)

• 한국환경성돌연변이발암원학회:학술대회논문집
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• 한국환경성돌연변이발암원학회 2004년도 KSOT/KEMS Fall Annual Meeting
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• pp.159-159
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• 2004