비글개에서 인체 재조합 적혈구 조혈인자, rHu-EPO의 급성독성에 관한 연구

조명행,성하정,김형식,곽승준,천선아,임소영,김원배,김병문,안병옥,이병무,Cho, Myung-Hang,Seong, Ha-Jung,Kim, Hyung-Sik,Kwack, Seung-Jun,Chun, Sun-Ah,Lim, So-Young,Kim, Won-Bae,Kim, Byoung-Moon,Ahn, Byoung-Ok,Lee, Byung-Mu 한국독성학회 1996 Toxicological Research Vol.12 No.2

The acute toxicity of rHu-EPO, newly developed recombinant erythropoietin, was tested in beagle dogs. rHu-EPO, when administered intravenously at 25, 000 IU/kg, did not cause any death. Also, rHu-EPO did not induce any change of body weight, food intake and clinical signs compared to controls. There were no significant changes in hematological, urine analysis and pathological examination. These results showed that rHu-EPO did not induce any remarkable toxic response and the $LD_50$ was greater than 25, 000 IU/kg in beagle dogs.

Molecular analysis of hprt mutation in B6C3F1 mice exposed to ozone alone and combined treatment of 4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone and/or dibuty1 phthalate for 32 and 52 weeks

조명행,김민영,김현우,박진홍,김준성,진화,문서현,유국종,강가미,김윤신,김영철,Hae Yeong Kim,이기호,조현선 대한수의학회 2004 Journal of Veterinary Science Vol.5 No.4

Potential toxicological interactions of 4-(N-methyl-Nnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and/or dibuthyl phthalate (DBP) on ozone were investigated after 32- and 52-wk exposures using hprt mutation assay. Male and female B6C3F1 mice exposed to ozone (0.5 ppm), NNK(1.0 mg/kg), DBP (5,000 ppm), and two or three combinations of these toxicants 6 h per day for 32- and 52-wk showed increases in the frequencies of TGr lymphocytes comparedto the control groups. Additive interactions were noted from two combination groups compared to the ozone alone in both sexes of 32- and 52-wk studies. The most common specific mutation type in the hprt genes of test materials-treated male and female mice was transversion with very few transition. The results indicate that such dominant transversion may be responsible for toxicityand combined exposure to ozone, NNK, and DBP induces additive genotoxicities compared to ozone alone. Potential toxicological interactions of 4-(N-methyl-Nnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and/or dibuthyl phthalate (DBP) on ozone were investigated after 32- and 52-wk exposures using hprt mutation assay. Male and female B6C3F1 mice exposed to ozone (0.5 ppm), NNK(1.0 mg/kg), DBP (5,000 ppm), and two or three combinations of these toxicants 6 h per day for 32- and 52-wk showed increases in the frequencies of TGr lymphocytes compared to the control groups. Additive interactions were noted from two combination groups compared to the ozone alone in both sexes of 32- and 52-wk studies. The most common specific mutation type in the hprt genes of test materials-treated male and female mice was transversion with very few transition. The results indicate that such dominant transversion may be responsible for toxicity and combined exposure to ozone, NNK, and DBP induces additive genotoxicities compared to ozone alone.

Comparative study of PM2.5-and PM10-induced oxidative stress in rat lung epithelial cells

조명행,최진혁,김준성,김영철,김윤신,정남현 대한수의학회 2004 Journal of Veterinary Science Vol.5 No.1

Accurate estimation of the exposure-response relationship between ambient urban particulate matters(PM) and public health is important for regulatoryperspective of ambient urban particulate matters (PM). Ambient PM contains various transition metals and organic compounds. PM10 (aerodynamic diameter less than 10 μm) is known to induce diverse diseases such aschronic cough, bronchitis, chest illness, etc. However, recent evaluation of PM2.5 (aerodynamic diameter less than 2.5 μm) against health outcomes has suggested that the fine particles may be more closely associated with adverse respiratory health effects than particles of larger size. Thisstudy was performed to evaluate PM2.5-induced oxidative stress in rat lung epithelial cell in order to provide basic data for the risk assessment of PM2.5. PM2.5 showedhigher cytotoxicity than PM10. Also, PM 2.5 induced more malondialdehyde (MDA) formation than PM10. In Hoechst 33258 dye staining and DNA fragmentation assay, apopotic changes were clearly detected in PM2.5 treated cells in compared to PM10. Expression of catalase mRNA wasincreased by PM2.5 rather than PM10. PM2.5 induced higher Mth1 mRNA than PM10. In pBR322 DNA treated with PM2.5, production of single strand breakage of DNAwas higher than that of PM10. In Western blot analysis, PM2.5 induced more Nrf-2 protein, associated with diversetranscriptional and anti-oxidative stress enzymes, compared to PM10. Our data suggest that PM2.5 rather than PM10 may be responsible for PM-induced toxicity. Additional efforts are needed to establish the environmental standard of PM2.5.

B6C3F1 mice exposed to ozone with 4-(N-methy1-N-nitrosamino)-1-(3-pyridy1)-1-butanone and/or dibuty1 phthalate showed toxicities through alterations of NF-kB, AP-1,Nrf2, and osteopontin

조명행,김민영,송경석,박건호,장승희,김현우,박진홍,진화,유국종,조현선,강가미,김영철 대한수의학회 2004 Journal of Veterinary Science Vol.5 No.2

Toxic effects of ozone, 4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK), and/or dibutyl phthalate (DBP)were examined through NF-κB, AP-1, Nrf2, and osteopontin (OPN) in lungs and livers of B6C3F1 mice. Electrophoretic mobility shift assay (EMSA) indicated that mice treated with combination of toxicants induced high NF-κB activities. Expression levels of p105, p65, and p50 proteins increased in all treated mice, whereas IκB activity was inhibited in NNK-,DBP-, and combination-treated ones. All treated mice exceptozone-treated one showed high AP-1 binding activities. Expression levels of c-fos, c-jun, junB, jun D, Nrf2, and OPNproteins increased in all treated mice. Additive interactions were frequently noted from two-toxicant combination micecompared to ozone-treated one. These results indicate treatment of mixture of toxicants increased toxicity through NF-κB, AP-1, Nrf2, and OPN. Our data could be applied to the elucidation of mechanism as well as the risk assessment of mixture-induced toxicity.

Cap-independent protein translation is initially responsible for 4(N-methylnitrosamino)-1-(3-pyridyl)-butanone(NNK)-induced apoptosis in normal human bronchial epithelial cells

조명행,Seo-Hyun Moon,Hyun-Woo Kim,김준성,Jin-Hong Park,Hwa Kim,Gook-Jong EU,Hyun-Sun Cho,Ga-Mi Kang,Kee-Ho Lee 대한수의학회 2004 Journal of Veterinary Science Vol.5 No.4

Evidences show that eukaryotic mRNAs can perform protein translation through internal ribosome entry sites (IRES). 5'-Untranslated region of the mRNA encoding apoptotic protease-activating factor 1 (Apaf-1) contains IRES, and, thus, can be translated in a cap-independent manner. Effects of changes in protein translation pattern through rapamycin pretreatment on 4-(methylnitrosamino)-1-(3-pyridyl)-butanone(NNK, tobacco-specific lung carcinogen)- induced apoptosis in human bronchial epithelial cells were examined by caspase assay, FACS analysis, Western blotting, and transient transfection. Results showed that NNK induced apoptosis in concentration- and timedependent manners. NNK-induced apoptosis occurred initially through cap-independent protein translation, which during later stage was replaced by cap-dependent protein translation. Our data may be applicable as the mechanical basis of lung cancer treatment.

Effects of cadmium on the gap junction, cell proliferation, protein kainase C activity and intracellular calcium levels in rat liver epithelial cells

조명행 강원대학교 종합약학연구소 1996 약학연구 Vol.9 No.-

비글개에서 인체 재조합 적혈구 조혈인자, rHuEPO의 아만성 정맥독성에 관한 연구

조명행,성하정,김형식,곽승준,천선아,한하수,임소영,안미영,김원배,김병문,안병옥,홍성렬,이병무 성균관대학교 약학연구소 1998 成均藥硏論文集 Vol.10 No.1

The subchronic toxicity study of rHuEPO, a newly developed recombinant erythropoietin, was investigated for 13 weeks in Beagle dogs intravenously treated with doses of 100,500 and 2,500 IU/㎏/day. There were no significant changes in body weight, food intake, physical and opthalmic examination, urine analysis, etc. Any toxic response was not observed except for enlarged spleen and extramedullary hematopoiesis. These results indicate that the no-observed adverse effect level (NOAEL) of rHuEPO is 100 IU/㎏ in Beagle dogs.

Azinphos-methyl이 랫트 태아에 미치는 기형학적 연구

조명행,이창업,이영순,Cho, Myung-Haing,Lee, Chang-Eop,Lee, Yong-Soon 한국독성학회 1988 Toxicological Research Vol.4 No.1

azinphos-methyl을 S.D.랫트에 투여하여 기형이 유발되는지의 여부와 태아의 기관형성 및 자궁내에서 태아의 발달에 미치는 영향을 알아보기 위하여 무처치의 negative control군, 수도물을 경구투여한 sham control군, 이미 기형효과가 있는 것으로 알려진 aspirin투여의 positive control군, 그리고 azinphosmethyl 0.094 mg/kg, 0.4 mg/kg 및 1.5mg/kg 투여군으로 나누어 각군 30마리씩으로, 임신 6~15일 사이에 경구투여한 결과는 다음과 같다. 즉, 모체의 체중증가율은 임신 7~14일, 즉 약물투여기에 aspirin 투여군과 azinphos-methyl 1.5mg/kg 투여군에서 현저한 감소를 보였다(p<0.01). 모체의 장기무게는 azinphos-methyl 1.5mg/kg 투여군에서 간장의 절대무게비가 유의성(p<0.05) 있는 감소를 보였으며, 신장의 절대 및 상대무게비는 aspirin군에서 (p<0.05, p<0.01), 또 난소의 절대 및 상대무게비는 aspirin 투여군(p<0.01)과 azinphos-methyl 전처치군(p<0.05)에서 유의성 있게 증가하였다. 모체 간장의 단백질량은 aspirin과 azinphos-methyl 1.5mg/kg 투여군에서 현저히 감소되었다(p<0.01). 모체의 배아와 태자에 대한 관찰 결과 azinphos-methyl 1.5mg/kg 투여군에서 암수의 비율에 있어 현저하게 증가(p<0.01) 했으며, 체중은 aspirin과 azinphos-methyl 1.5mg/kg 투여군에서 고도의 유의성(p<0.01) 있게 감소하였고, azinphos-methyl 0.4 mg/kg 투여군에서도 현저하게 감소하였다(p<0.05). 그리고 미숙태자와 흡수태자는 aspirin군에서, 죽은 태자수는 azinphos-methyl 1.5mg/kg 투여군에서 유의성 있게 증가하였다(p<0.05, p<0.01). 태자의 장기에서 나타난 기형은 aspirin과 azinphos-methyl 1.5mg/kg 투여군 공히 횡경막에서 횡격막 천공, 두부에서 무안구증, olfactory bulb의 확장, 수두뇌, 그리고 3뇌실과 측뇌실의 결손, 심장에서 좌심실벽 위축, 심첨확장 등이 유의성(P<0.01, P<0.05) 있게 관찰되었으며, 특히 횡경막, 심장 및 안구등에 높은 기형발생율을 보였다. 또 태자의 골화지연은 두개골에서 후두골, 접형골, 구개, 흉골에서 4번째 흉골편, 검상돌기, 척추에서 경추, 흉추, 미추, 전${\cdot}$후지골, 중족골에서 관찰되었다. 한편, 자연 분만시킨 태자의 사망율은 azinphos-methyl 1.5mg/kg 투여군에서 negative control군에 비해 고도의 유의성(p<0.01) 있게 높음을 알 수 있었으며, 체중도 역시 현저하게(p<0.01) 감소하였고, 발육지표중 pivoting, 체모형성, 청각능력, 시각능력, 사지근육 발달정도 및 고환하강시기 등에서 고도의 유의성(p<0.01)이 관찰되었다. This study was carried out to investigate the teratological potential of azinphos-methyl in the rat fetuses and to establish the nature of the effects on organogenesis and intrauterine development. The Sprague-Dawley female rats (180-210g) without previous litter were used in this study. Azinphos-methyl dosages of 0.094mg/kg, 0.4mg/kg, 1.5mg/kg were selected based on the acute intragastric $LD_{50}$ of 15mg/kg in the rat. Azinphos-methyl in water (Treatment Group), non-treatment control (Negative Control), water control (Sham Control), were administered by oral route and aqueous solution of acetyl salicylic acid (Positive Control) was administered by gavage at rate of 10 ml/kg of body weight from day 6 through 15. The results obtained were summarized as follows. 1. Decreased body weight of dams was observed in animals treated with aspirin and azinphos-methyl 1.5 mg/kg from day 7 through 14. (P<0.01) 2. There was an apparent decrement in the absolute liver weight in the azinphos-methyl 1.5 mg/kg treated group (P<0.05). However, the absolute and relative kidney weight in aspirin group (P<0.05, P<0.01) and the absolute and relative ovary weight in aspirin, azinphos-methyl treatment groups (P<0.01, P<0.05) were increased. 3. Decreased protein contents of dam's liver was observed in the aspirin and high dose azinphos-methyl treated group of animals (P<0.01). 4. The number of male-female ratio per dam increased in azinphos-methyl 1.5 mg/kg group but there was an apparent decrement in the body weight of fetuses in aspirin and high dose azinphos-methyl group (P<0.01, P<0.05). Total immature and resorbed fetuses were increased in aspirin group and the number of dead fetuses were also increased in azinphos-methyl 1.5mg/kg treated group of animals. (P<0.01, P<0.05). 5. In soft tissue defects, diaphragmatic hernia in diaphragm, anophthalmia, enlarged olfactory bulb, hydrocephalus, absence of third and lateral ventricle in skull, hydronephrosis in kidney, atrophy of left ventricle wall, enlarged apex in heart were observed. Especially, defects of diaphragm, heart and eye ball showed peak incidences in the high dose azinphosmethyl and aspirin group. (P<0.01). 6. Variations in the ossification patterns of skull, sternebrae, tail, forelimbs and hindlimbs showed peak incidences in the aspirin and high dose azinphos-methyl group. (P<0.01). 7. In the developmental indices of offspring, the mortality of aspirin and azinphos-methyl 1.5mg/kg treated group was higher than that of negative control. And, there was an apparent decrement in the body weight of fetuses (P<0.01) and considerable differences were obtained in pivoting, development of fur, auditory function, vision, quadrupled muscle development and testes descent in aspirin and azinphos-methyl 1.5mg/kg group. (P<0.01).

비글개에서 인체 재조합 적혈구 조혈인자 , rHuEPO 의 아만성 정맥독성에 관한 연구

조명행(Myung Haing Cho),성하정(Ha Jung Seong),김형식(Hyung Sik Kim),곽승준(Seung Jun Kwack),천선아(Sun Ah Chun),한하수(Ha Su Han),임소영(So Young Lim),안미영(Mi Young Ahn),김원배(Won Bae Kim),김병문(Byoung Moon Kim),안병옥(Byoung Ok 한국응용약물학회 1998 Biomolecules & Therapeutics(구 응용약물학회지) Vol.6 No.3

The subchronic toxicity study of rHuEPO, a newly developed recombinant erythropoietin, was investigated for 13 weeks in Beagle dogs intravenously treated with doses of 100, 500 and 2,500 IU/㎏/day. There were no significant changes in body weight, food intake, physical and opthalmic examination, urine analysis, etc. Any toxic response was not observed except for enlarged spleen and extramedullary hematopoiesis. These results indicate that the no-observed adverse effect level (NOAEL) of rHuEPO is 100 IU/㎏ in Beagle dogs.

오줌유래 Trypsin 효소 억제제가 췌장염에 미치는 영향에 관한 연구

조명행(Myung Haing Cho),권오경(Oh Kyung Kweon),정요찬(Yo Chan Jeong),유아선(Ah Sun You),김종민(Jong Min Kim),박수진(Soo Jin Park),송동호(Dong Ho Song) 한국응용약물학회 1996 Biomolecules & Therapeutics(구 응용약물학회지) Vol.4 No.3

The metabolism of carbinoxamine, 2-[(4-chlorophenyl)-2-pyridinyl-methoxy]-N, N-dimethylethaneamine, was studied in adult male volunteers after an oral dose of 15 mg. Solvent extracts of urine obtained with or without enzyme hydrolysis were analyzed by gas chromatography-mass spectrometry after derivatization with MSTFA/TMSCl (N-methyl-N-trimethylsilyltrifluoroacetamide/trimethyl chlorosilane). The structures of metabolites were determined based on the electron impact (EI) and chemical ionization (CI) mass spectra. Nonconjugated metabolites identified in the urine were carbinoxamine, nor-carbinoxamine, and bis-nor-carbinoxamine. Parent drug, nor-carbinoxamine, and bis-nor-carbinoxamine were also detected as conjugated forms. These metabolites observed in human urine were different from those previously reported in the rat. Urinary excretions of carbinoxamine were reached to maxima in 4 hours after drug administration with 4.9%-8.1% and 2.5-4.2% of the dose excreted during 24 h as carbinoxamine and its glucuronide, respectively.