• Archives of Pharmacal Research
• /
• 제15권1호
• /
• pp.78-86
• /
• 1992

The induction of hepatic cytochromes P450 and metabolic effects have been examined in male and female Sprague-Dawley rats following treatment with either phenobarbital or 3-methylcholanthrene. Hepatic cytochrome P450 levels were higher in males than in females by ~40%. Treatment of male and female rats with phenobarbital or 3-methylcholanthrene resulted in an ~1.6 and 2-fold increase, respectively, in heptic microsomal cytochrome P450 levels in both sexes, relative to untreated animals. Immunoblot analyses were performed to compare sex-related changes in P450 levels. Hepatic P45IIB1 levels in males were greater than those in females following phenobarbital treatment. 3-Methylcholanthrene-induced male hepatic microsomes exhibited greater levels of P450 females failed to exhibit a band. Mab PCN 2-13-1 against P-45-IIIA recognized an intense in uninduced microsomes from female rats. The levels of P450IIIA in males were increased 2 to 3-fold following treatment with phenobarbital, while the increase of IIIA levels in females by phenobarbital was minimal, as monitored by immunoblot analysis. Solid phase radiommunoassay using monoclonal antibodies supported the results of immunoblot analysis. Phenobarbital treatment caused a 6.5-fold increase in the monoclonal iantibody binding to IIBI in males, whereas treatment of females with phenobarbital resulted of IA levels by 3-methylcholanthrene was also greater in females than in males (10-vs. 8-fold) although the levels of induced IA were comparable inboth sexes, as assessed by radiommunoassay. Radioimmunoassay also showed that hepatic IIEI level was 1.5-fold higher in males than in females and that either phenobarbital or 3-methylcholanthrene treatment caused 80% to 40% decrease in IIEL levels, relative to control, in both sexes. Sex-related metabolic activities were examined in hepatic microsomes. Hexobarbital hydroxylase activity was 2-to 3-fold higher in uninduced microsomes from males than that from females. This hydroxylase activity was increased 2-and 3-fold in males and females, respectively, following phenobarbital treatment, as compared to controls. Addition females produced 64% and 84% inhibition of hexobarbital oxidation, respectively. Aryl hydrocarbon hydroxylase activity was increased -12 and 26-fold in males and females respectively. Following phenobarbital treatment, as compared to controls. Addition of anti-P450IIB1 antibody to phenobarbital-induced hepatic microsomes from males and females produced 64% and 84% inhibition of hexobarbital oxidation, respectively. Aryl hydrocarbon hydroxylase activity was increased -12 and 26 fold in males and females, respectively, following 3-methylcholanthrene treatment relative to controls. The anti-P-450IA antibody inhibitable rate of aryl hydrocarbon hydroxylase activity was comparable in both sexes following 3-methylcholanthrene treatment relative to controls. The anti-P450LA antibody inhibitable rate of aryl hydrocarbon hydroxylase activity was comparable in both sexes following 3-methylcholanthrene treatment (-70%). These results demonstrate that levels of hepatic P450IIB1 or P450IA are greater in male than in female for untreated, phenobarbital-or-3-methylcholanthrene treated rats. In addition, the relative for untreated phenobarbital-or 3-methylcholanthrene treated rats. In addition, the relative increase of phenobarbital-or 3-methylcholanthrene treated rats. In addition, the relative increase of phenobarbital-or 3-methylcholanthrene treated rats. In addition, the relative increase of P450IIB1 or IA phenobarbital or 3-methylcholanthrene is more significant in females.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 1992년도 제1회 신약개발 연구발표회 초록집
• /
• pp.55-55
• /
• 1992

다환상 방향족탄화수소류인 3-methylcholanthrene 이 자궁에 미치는 영향을 연구하고자 자궁의 성장에 미치는 영향과 에스트로겐 수용채와의 상호작용을 조사하였다. 3-Methylcholanthrene 단독 투여군은 대조군에 비해 투여 용량에 관계없이 유의성있는 자궁 무게의 변화를 나타내지 않았으나 diethylstilbesterol과 3-methylcholanthrene 병용 투여군에서는 diethylstilbesterol 단독 투여군에 비해 자궁 무게가 병용 투여한 3-methylcholanthrene 용량에 의존적으로 감소하였다. 안티에스트로갠인 tamoxifen이 자궁 성장에 미치는 영향을 관찰하기 위하여 tamoxifen단독 투여시에는 자궁 무게가 대조군에 비해 약간 증가하여 (p<0.05)부분 효능 작용을 나타냈으나 diethylstilbesterol과 동시 투여시에는 diethylstilbesterol에 의한 자궁 무게 증가가 감소되었으며 그 감소 정도는 병용한 tamoxifen 용량에 의존적이었다. Diethylstilbesterol과 3-methylcholanthrene 및 tamoxifen을 병용 투여했을때 diethylstilbesterol에 3-methylcholanthrene만을 병용 부여하였을 때보다 diethylstilbesterol에 의한 자궁 성장이 유의적으로 (P<0.01)감소되었다. Diethylstilbesterol과 3-methylcholanthrene의 병용 투여군에서는 투여한 후 시간이 경과함에 따라 diethylstilbesterol 단독 투여군에 비해 일정한 비율(23.6=3.9％)로 자궁 무게 증가가 억제되었다.

• Archives of Pharmacal Research
• /
• 제20권5호
• /
• pp.404-409
• /
• 1997

Trout CYP1A-CAT expression construct was generated by cloning -3.5 Kb $5^I$ flanking DNA of trout liver CYP1A gene in front of CAT gene at pCAT-basic vector. Hepa 1 cells, which are known to contain a functional arylhydrbcarbon $receptor^I$ were transfected with trout CYP1A-CAT using lipofectin. 3-Methylcholanthrene (1 nM) was added into hepa 1 cells in culture in order to examine if $5^I$ flanking DNA of trout CYP1A gene could interact with mouse transactivating factors to bring about transcription of the chloramphenicol acetyltransferase(CAT) reporter gene. The level of CAT protein was measured by CAT ELISA and the level of CAT mRNA was determined by RTPCR. The treatment of 1 nM 3-methylcholanthrene resulted in two fold increases in CAT protein as well as CAT mRNA compared to untreated control hepa 1 cells. These data indicate that arylhydrocarbon receptors of mouse hepa 1 cells are functional to activate exogenously transfected trout CYP1A-CAT construct in terms of both transcription and translation of CAT. We also examined the effect of 3-methylcholanthrene on endogenous cyplal activity in hepa 1 cell. 3-Methylcholanthrene (1 nM) treatment to hepa 1 cells trahsfected with trout CYP1A-CAT construct stimulated the level of cyp1a1 mRNA by two folds and the activity of ethoxyresorufin-O-deethylase by two fold compared to that of control cells. In this study we reported that trout CYP1A-CAT reporter gene expression construct could be expressed by 3-methylcholanthrene treatment in mouse hepa 1 cells. Thus trout CYP1A-CAT could serve as a good model to study the mechanism of regulation of CYP1A1 gene expression.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
• /
• pp.192-192
• /
• 1994

본 논문에서는 cytochrome P450 LA1 유전자의 5'-upsteam 조절부위의 클로닝을 실시하였다. pUC19 vector에 연결시킨 3.4 Kb 크기의 Pstl DNA조각을 Sst1, Nco1 제한 효소로 자른 뒤, Exonuclease III 를 처리하여 약 200bp 씩의 차이를 갖는 여러 크기의 plasmid들을 얻었다. 이 plasmid 의 핵산서열을 알아보기 위해 dideoxy nucletide를 이용한 sequencing방법으로 그 핵산서열의 결정 실험을 시도하였다. 또한, 다환상 방향족 탄화수소 화합물에 반응성을 갖는 C57BL/6N 생쥐와 반응성을 갖지않는 DBA/2N 생쥐에 있어 phase II 대사 효소인UDP-glucuronosyltransferase 효소활성에 대한 3-methylcholanthrene의 영향을 알아보기 위해 C57BL/6N 생쥐와 DBA/2N 생쥐에 각각 다른 농도의 3-methylcholanthrene을 처리하거나 각기 다른 시간에 3-methylcholanthrene를 처리하였다. 그 결과 UDP-glucuronosyl-transferase의 mRNA가 3-methylcholanthrene양의 증가에 따라, 처치시간이 길어짐에 따라 증가되어지며 그 mRNA위 크기는 약 2.2Kb 정도임을 알았다. 이로부터 UDP-ghucuronosyltransferase 또한 cytochrome P45O와 함께 다환상 방향족 탄화수소 화합물 조절인자를 통한 조절을 받을 것이며 phase I phase II 약물 대사 효소가 조절상 밀접한 관련을 가짐을 예측할 수 있었다.

• Toxicological Research
• /
• 제17권2호
• /
• pp.123-129
• /
• 2001

This study were carried out to investigate cytotoxicity of paraquat and bentazon that is scattering to farm products were essensial for human diet and compensatory effects of 3-methylcholanthrene (3-MC) in vitro and in vivo. In vitro, The 5.0$\times$$10^4$ cell/ml of NIH 3T3 fibroblast in each well of 24 multidish were cultured. After 24 hours, the cells were treated with solution of paraquat and bentazon (1, 25, 50, 100 pM respectively). After the NIH 3T3 fibroblast of all groups were cultured in same condition for 48 hours, Sulfohordamin B Protein (SRB) assay were performed to evaluate the cytotoxicity of cell organelles. Paraquat and bentazon $SRB_50$ were 1860.73 $\mu\textrm{M}$, 1913.38 $\mu$M respectively. In vivo, Sprague Dawley male rats divided into paraquat and bentazon only administered group and simultaneous application group of paraquat and bentazon and 3-MC. At 30 min. and 1, 3, 6, 12, 24, 48 and 96 hrs. interval after each treatment, the animals were sacrificed by decapitation and kidney were immediately removed, immersed in fixatives, and processed with routine method for light microscopic study. Paraffin sections were stained with H-E, PAM, and PAS. Under the light microscope, atrophic change of renal corpuscles were frequently observed from 3 hrs after paraquat and bentazon treatment. The increase of the mesangium was apparent from 12 hrs later after paraquat and bentazon treatment. Necrotic changes of the epithelium and loss of brush border of proximal tubules were most severe at 48 hrs after paraquat and bentazon treatment, respectively. In contrast there were no evidences of the toxic effects on renal tissues at 48hrs in paraquat and bentazon plus 3-MC treated groups.

• Archives of Pharmacal Research
• /
• 제16권4호
• /
• pp.276-282
• /
• 1993

This study has been undertaken to examine the effect of 3-methylcholanthrene (3MC) on rat uterine growth and to understand the mechanism of action of 3MC in rat uterus. After diethylstilbesterol(DES) or tamoxifen(TAM) or 3MC or DES plus TAM or DES plus 3MC was administered into immature female rats, uterine weight over corn oil-treated uteri. 3MC treatment had no effect on uterine weight but, DES stimulated uterine weight was inhibited by 3MC concomitant tratment. While TAM alone treatment showed slight increase in uterine wieght, inhibited uterine growth simulated by DES when it was adiministrated with DES condirect binding assay with $[^3H]$ estradiol and the relative binding affinities of 3MC and TAM were estimated by competetion assy. Estradiol tumed out to have high affinity for rat uterine estrogen receptor (kd = 0.4 nM). The relative binding affinities of TAM and 3MC were 1% and 4.7% that of DES for rat uterine estrogen receptor, respectively. 3MC was shown to have similar affinity for eat uterine estrogen receptor to that of TAM. Effects of DES 3MC and TAM administration in vivo on rat uterine estrogen recptor level were examined. It was confirmed that the estrogen, DES and antiestrogen, TAM decreased estrogen receptor levels from rat ulterus and also 3MC decreased rat uterine estrogen receptor level when rats were treated with DES, TAM and 3MC in vivo. Data indicates that 3MC acts as an antiestrogen mediated through estrogen receptor system.

• 한국환경농학회지
• /
• 제21권1호
• /
• pp.57-68
• /
• 2002

쥐에서 carbofuran 대사에 미치는 phenobarbital sodium (PB) 또는 3-methylcholanthrene (3-MC)의 영향을 조사하기 위하여 쥐에 이들을 단독 또는 조합으로 경구투여한 후 일정 간격으로 쥐의 주요 장기, 대변, 소변 및 혈액 중 대사산물의 종류와 생성율을 조사하였다. Carbofuran 단독투여와 carbofuran과 PB 또는 3-MC 조합투여 모두 경구투여 후 48시간 이내에 빠르게 배설되어 $^{14}C$-carbofuran 총 투여량의 79.9$\sim$81.1%가 소변으로, 5.7$\sim$6.5%가 대변으로 배설되었는데, 배설속도는 carbofuran 단독투여 보다 carbofuran과 PB 또는 3-MC 조합투여에서 빨랐다. 쥐의 주요 장기, 대소변 및 혈액 중의 carbofuran의 대사산물은 공통적으로 3-hydroxycarbofuran, 3-ketorarbofuran, 3-hydroxycarbofuran phenol, 3-ketocarbofuran phenol과 carbofuran phenol이었고, 주요 대사산물은 3-hydroxycarbofuran과 3-ketocarbofuran이었는데, 주요 대사산물의 경우 carbofuran만의 투여에서는 3-hydroxycarbofuran이었으나 carbofuran과 PB 또는 3-MC 조합투여는 3-ketocarbofuran이었다. 소변 중 carbofuran의 2가지 주 대사산물의 생성율은 carbofuran 단독투여시 3-hydroxycarbofuran 17.4%와 3-ketocarbofuran 12.8%이었고, carbofuran과 PB 또는 3-MC 조합투여시 3-hydroxycarbofuran 8.6%와 3-ketocarbofuran 23.5%로서, carbofuran 단독투여와 carbofuran과 PB 또는 3-MC 투여사이에 대사산물의 종류는 같았으나 생성율에는 큰 차이가 있었다. 이와 같은 결과는 쥐에 carbofuran 투여 후 PB나 3-MC를 투여함으로써 carbofuran의 대사가 빠르게 이루어지고, 주 대사산물 중3-hydroxycarbofuran보다 독성 이 낮은 3-ketocarbofuran으로의 대사가 빠르게 이루어지기 때문에 carbofuran의 독성이 경감되어 쥐가 생존할 수 있는 것으로 판단된다.

• Applied Biological Chemistry
• /
• 제39권1호
• /
• pp.77-83
• /
• 1996

이스라엘 잉어에 있어서 carbamate계 살충제 carbofuran의 독성에 미치는 phenobarbital sodium(PB) 또는 3-methylcholanthrene (3-MC)의 영향과 작용기작을 효소적 측면에서 구명할 목적으로 carbofuran과 PB나 3-MC를 이스라엘 잉어에 각 조합으로 처리하여 독성경감 효과를 조사하였고, 공시한 농약과 PB나 MC가 acetylcholinesterase(AChE), glutathione S-transferase(GST), UDP-glucuronosyltransferase(UDPGT) 및 cytochrome P-450-dependent monooxygenase(monooxygenase)의 효소활성에 미치는 영향을 조사하기 위하여 carbofuran과 PB나 3-MC를 각각 조합으로 처리한 후 경시적으로 이스라엘 잉어의 각 효소들의 상대활성도를 조사하였다. PB와 3-MC만 투여한 실험군에서 이스라엘 잉어의 생존수는 무처리군과 동일하였고 살충제만 처리한 실험군의 이스라엘 잉어 생존수는 처리농도가 증가하면서 감소되었으나, PB나 3-MC와 살충제를 조합처리한 실험군에서는 살충제만 처리한 실힘군에 비하여 매우 높은 생존율을 나타낸 것으로 보아 해독효과가 인정 되었다. 효소활성(in vivo)은 AChE의 경우 carbofuran 0.95 ppm만을 처리한 실험군에서는 24시간내내 각 조사시기마다 무처리군에 비해 40% 이상의 활성저해를 보였으나 carbofuran과 PB 및 3-MC를 조합처리한 실험군에서는 효소활성이 초기에 감소하다가 서서히 증가하여 24시간후에는 무처리군과 비슷한 수준을 나타냈고, GST의 경우 carbofuran만을 처리한 실험군에서는 초기에 약 20% 이상의 활성저해를 보였으나 carbofuran과 PB나 3-MC를 조합처리한 실험군에서는 약제처리 1시간 후 부터 무처리군에 비해 효소활성이 20% 이상 증가하였다. UDPGT와 monooxygenase의 효소활성은 carbofuran과 PB나 3-MC를 조합처리한 실험에서 처리 $6{\sim}12$시간 후에는 carbofuran 처리군에 비해 효소활성이 $4{\sim}8$배 이상 급격히 높아졌다. 이상의 결과에서 PB 및 3-MC처리가 이들 효소의 활성을 유도함으로써 carbofuran의 독성으로 부터 이스라엘 잉어를 보호 하는데 관여한 것으로 보인다.

• 농약과학회지
• /
• 제3권3호
• /
• pp.27-36
• /
• 1999

쥐에 있어서 carbamate계 살충제 carbofuran의 독성에 미치는 phenobarbital sodium(PB) 또는 3-methylcholanthrene(3-MC)의 영향과 작용기작을 효소적 측면에서 구명할 목적으로 이들을 단독 또는 조합으로 경구투여 하여 in vivo 효소활성을 조사하였다. Acetylcholinesterase(AChE)와 butyrylcholinesterase(BuCheE)의 효소활성은 carbofuran 3.8 mg/kg을 투여하였을 때 48시간까지 $20{\sim}70%$ 범위의 저해를 보였고, carbofuran과 PB 또는 3-MC를 조합투여하였을 때 효소활성은 초기에 감소하다가 24시간 후에는 대조구와 비슷한 수준을 나타냈다. Glutathione S-transferase(GST)의 경우 carbofuran만을 투여하였을 때 초기($0.5{\sim}6$ hr)에 $15{\sim}35%$의 저해를 보였으나, carbofuran과 PB 또는 3-MC의 조합투여시 초기에는 약간 저해를 보이다가 3시간 후에는 대조군과 유사한 효소활성을 보였고, 6시간 후에는 대조군에 비해 활성이 20%이상 증가하였다. UDP-glucuronosyltransferase(UDPGI) 및 cytochrome P-450 효소계의 효소활성은 carbofuran과 PB 또는 3-MC를 조합 투여하였을 때 투여 후 6시간까지는 carbofuran만의 투여에 비해 효소활성이 $2.6{\sim}2.8$배 이상 높았다. 이상의 결과에서 PB 및 3-MC의 투여가 이들 효소활성을 유도하므로써 carbofuran의 독성으로부터 쥐를 보호한 것으로 판단된다.

• 대한수의학회지
• /
• 제26권1호
• /
• pp.109-115
• /
• 1986

Experiments were performed on mice to investigate the effects of a polycyclic aromatic hydrocarbon, 3-methylcholanthrene (MCA) on Arthus reaction, delayed-type hypersensitivity (DTH) and antibody production to sheep red blood cells (SRBC). Mice were sensitized iv with 0.1ml of 1% SRBC suspension were treated with a single ip injection of olive oil alone or with different doses of MCA in oil (0.5~50mg/Kg) at various time before (-) or after (+) sensitization (day 0) and were challenged at 4 days after SRBC. Arthus reaction was measured at 3 hours after challenge and other responses at 24 hours. Treatment with MCA inhibited Arthus reaction and DTH to SRBC, measured by footpad swelling reaction, and this immunosuppressing effect was dependent on the dose and time of MCA treatment in relation to SRBC sensitization. Humoral immune responses as measured by serum hemagglutinin-and hemolysin-titers to SRBC were significantly depressed when MCA was injected before or at the same time of sensitization. However, the response was slightly depressed when injected after SRBC. These results indicate that MCA suppress the function of the cells involved in immune responses.