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Vitamin A 와 Ethanol 이 Dimethylnitrosamine 에 의한 Mouse 조직내의 DNA , RNA 및 단백질의 손상도에 미치는 영향
김재현,홍연탁,박정식,김승희,강태규,홍성렬,박창원,권오철,이동권 ( Jea Hyun Kim,Youn Tak Hong,Jung Sik Park,Seung Hee Kim,Tea Gyu Kang,Sung Roul Hong,Chang Won Park,O Cheol Kweon,Dong Kwon Rhee ) 생화학분자생물학회 1991 BMB Reports Vol.24 No.6
Dimethylnitrosamine(DMN) has been shown to induce tumors of the liver, kidney, esophagus and lung. Furthermore low levels of DMN (0.1∼1 ppb) have been detected in normal human blood as well as urine. The purpose of this study is to determine extent of the covalent binding of DMN to DNA, RNA and protein in ethanol- and vitamin A- pretreated mouse. Ethanol or vitamin A was administered for 3 weeks and ^(14)C-labeled DMN was administered intraperitoneally. Then DNA, RNA and protein were isolated from liver, brain and pancreas and used for determination of radioactivity. DMN was bound selectively to liver DNA rather than brain or pancreas DNA. DNA damage in liver was 1.4 and 1.7 times greater than protein and RNA damage, respectively. Ethanol pretreatment and vitamin A pretreatment increased DNA damage in brain significantly upto 6.7 and 9.5 times, respectively, than the control group.
Vitamin A와 Ethanol이 Dimethylnitrosamine에 의한 Mouse조직내의 DNA, RNA 및 단백질의 손상도에 미치는 영향
김재현,홍연탁,박정식,김승희,강태규,홍성렬,박창원,권오철,이동권,Kim, Jea-Hyun,Hong, Youn-Tak,Park, Jung-Sik,Kim, Seung-Hee,Kang, Tea-Gyu,Hong, Sung-Roul,Park, Chang-Won,Kweon, O-Cheol,Rhee, Dong-Kwon 생화학분자생물학회 1991 한국생화학회지 Vol.24 No.6
Dimethylnitrosamine(DMN)은 간 및 폐, 신장, 식도 등에 앙을 유발시키는 물질이며 정상인의 혈액 및 뇨에서도 0.1~1 ppb정도가 검출되고 있다. DMN은 DNA를 methylation시켜 비정상적인 adduct를 형성하여 암을 유발하게 된다. 본 연구는 ethanol 또는 vitamin A를 mouse에 전처리 하였을 때 $[^{14}C]$으로 표지된 DMN이 생체내의 DNA, RNA 및 단백질과 어느 정도 공유결함을 형성하여 손상을 주는지 측정하였다. 실험결과 DMN은 뇌 또는 췌장에서 보다는 간에 선택적으로 결합하였으며 이 때 간의 DNA와 결합하는 것이 단백질 또는 RNA와 결합하는 것보다 각각 1.4배와 1.75배 높았다. Ethanol을 전처리 하였을 때는 DMN이 ethanol을 투여하지 않은 군보다 간의 경우 RNA와 단백질이 각각 7.5배, 1.4배 정도 더 결합하였다. Ethanol을 전처리한 뇌의 DNA에 6.7배 많이 결합하였으나 RNA에는 오히려 대조군의 15.9% 정도로 크게 감소되었다. 또한, 췌장의 단백질에는 2.0배, RNA의 경우 1.7배 정도 각각 더 결합한 결과를 나타내었다. 반면에 vitamin A 전처리군에서 간 단백질의 adduct는 35% 유의성 있게 감소되었으나 뇌 DNA에서 는 유의성 있게 9.5배 증가하였다. Dimethylnitrosamine(DMN) has been shown to induce tumors of the liver, kidney, esophagus and lung. Furthermore low levels of DMN (0.1~1 ppb) have been detected in normal human blood as well as urine. The purpose of this study is to determine extent of the covalent binding of DMN to DNA, RNA and protein in ethanol- and vitamin A- pretreated mouse. Ethanol or vitamin A was administered for 3 weeks and $^{14}C$-labeled DMN was administered intraperitoneally. Then DNA, RNA and protein were isolated from liver, brain and pancreas and used for determination of radioactivity. DMN was bound selectively to liver DNA rather than brain or pancreas DNA. DNA damage in liver was 1.4 and 1.7 times greater than protein and RNA damage, respectively. Ethanol pretreatment and vitamin A pretreatment increased DNA damage in brain significantly upto 6.7 and 9.5 times, respectively, than the control group.
Quinone계 화합물의 발암성 조기검색법에 관한 연구
조대현,홍진태,박정식,홍연탁,진강,정명희,이병무,Cho, Dae-Hyun,Hong, Jin-Tae,Park, Jeong-Sik,Hong, Youn-Tack,Chin, Kang,Jung, Myung-Hee,Lee, Byung-Mu 한국독성학회 1992 Toxicological Research Vol.8 No.2
To investigate a short term screening method for carcinogenic quinone compounds, 8-hydroxydeoxyguanosine (8-OHdG), an oxidative DNA damage, was determined in the kidney and liver DNA isolated from Sprague-Dawley rats after i.p.injection of 7 mg/kg adriamycin (AM), 7mg/kg tetrahydropyranyladriamycin (THP), and 10mg/kg daunomycin (DM) by HPLC-electrochemical detector system. 8-OHdG was also determined from rat hepatocvtes and calf thymus DNA exposed to AM, DM and THP. When rats were treated with DM and THP, 8-OHdG was significantly increased in the kidney compared to control group, and remained at high level (7.9~9.0, 8-OHdG/dG${\times}10^4$)at the end of experiments (48hr after treatment). 8-OHdG level in cultured hepatocyte exposed to AM, DM and THP was 1.5~2 fold higher than control at all time points. (1,2,3,4hr after treatment). From calf thymus DNA exposed to AM, DM and THP, 8-OHdG was 2.5 fold higher than of control. These results suggest that quantitation of 8-OHdG may provide a useful marker for identifying target organ in oxidative chemical carcinogenesis and for short term screening of free radical generating carcinogens.
김재현,박창원,박정식,홍연탁 한국응용약물학회 1993 Biomolecules & Therapeutics(구 응용약물학회지) Vol.1 No.1
DNA adducts induced by putative chemical related to carcinogenesis were detected and determined by (32)^P-Postlabelling assay after exposure of 4 compounds comprising two azo dyes (amaranth, new coccine) and two flavonoid compounds (rutin, quercetin) to ICR mouse. DNA was isolated from mouse liver and digested enzymatically to deoxyribonucleoside 3'-monophosphate. The postincubation of DNA digests with nuclease P1 before (32)^P-labelling enhanced the technique's sensitivity. Nuclease P1 cleaves deoxyribonucleoside 3'-monophosphates of normal nucleotides to deoxyribonucleosides which do not serve as substrates for polynucleotide kinase, while most of adducts were found to be totally or partially resistant to the 3'-dephosphorylating action of nuclease Pl. The adducted deoxyribonucleoside 3'-monophosphate was converted to 5'-(32)^P-labelled deoxynucleoside 3',5'-bisphosphate by T4 polynucleotide kinase. The nucleotides were separated by anion-exchange thin layer chromatography(TLC) on polyethyleneimine cellulose by 4-dimensional or 2-dimensional TLC then detected by autoradiography. The results show that DNA adducts were detected in liver DNA of ICR mouse after administration of amaranth and quercetin by 2-dimensional and/or 4-dimensional TLC.