2000 Fiscal Year Annual Research Report

生体内酵素反応解析用光ファイバセンサシステムの開発とその発癌機構解明への応用

Research Project

Project/Area Number	11650431
Research Institution	Hokkaido Institute of Technology
Principal Investigator	佐々木一正北海道工業大学, 工学部, 教授 (00002006)
Co-Investigator(Kenkyū-buntansha)	数坂昭夫北海道大学, 大学院・獣医学研究科, 助教授 (00002113)
Keywords	代謝酵素 / チトクロムP450 / 光ファイバセンサ / 発癌物質 / 癌発症機構 / プラスチック光ファイバ / CYP1A1
Research Abstract	本研究は、光ファイバセンサを用いて生体中の酵素をin vivoで検出する新技術を開発し、これを発癌機構の解明へ応用することを目的に、北海道工業大学と北大獣医学研との共同研究として主に次のような2つのプロジェクトとして進められた。 (1)ファイバセンサシステムの開発と装置の試作であり、主に、北海道工業大学において行った。 (2)試作されたシステムを用い、このシステムの発癌機構解明への適用について、北大獣医学研において行なった。その結果、(1)の成果として、これまでにない新しい酵素検出システム(Ver.1)を構築することに成功した。本装置を用いて、発癌物質であるbnzopyrenなどを投与したラットの肝臓におけるCYP1A1の発現を観察したところ、日を追って、酵素活性が高まっていく状況、発癌物質の濃度による差異などを高感度に検出することに成功した。また、controlにおいても微量な酵素による反応も捉えることができた。本装置では、反応の進捗を実時間で追跡することが可能であり、酵素反応機構を克明に解析できるようになった。また、(2)の成果として、発癌を引き起こすと考えられている要因として紫外線暴露をラットの耳表皮に与えたときのCYP1A1の発現状況とその知見に基づく、発癌の確率、治癒率などを定量的に計測評価できる見通しを得た。さらに、(1)で開発されたシステムに改良を加え、さらにパフォーマンス100程度改善したシステムの開発に成功した。これらの成果は、酵素の動態に関する知見が癌の発症前に発癌の診断に使えるという可能性を示したものであり、本システムは癌診断装置という方向で応用できることを意味するものである。

Research Products
(27 results)

All Other

All Publications (27 results)

[Publications] 吉田勝仁,長牛英雄,佐々木一正,沈艶波,数坂昭夫他2名: "生体用光ファイバセンシングシステムの試作と評価"社団法人電子情報通信学会技術報告集. OFT2000-8. 47-52 (2000)
[Publications] 吉田勝仁,長牛英雄,佐々木一正,沈艶波,数坂昭夫: "光ファイバセンサ方式による代謝酵素CytochromeP450センシングシステムの開発と特性"北海道工業大学研究紀要第29号. 第29号. 1-6 (1999)
[Publications] IsseiSasaki,AyakoKawanabe,Isao YamakawaAkio Kazusaka: "In vivo Observation of Cytochrome P4501A Isozyme Reaction in Liver of Living Rats by Using Fiber-Optic Sensor"Conference Proceedings of 13^<th> International Conference on Optical Fiber Sensor. 179-182 (1999)
[Publications] KazusakaA,NakamraA,HirotaT,MorinoA,ImaokaS,FunaeY,YamamotoY: "Cytochrome P-450isoforms responsible for the N-demethylation and cyclohexene-hydroxylation of NS-21"Xenobiotica. 29. 243-252 (1999)
[Publications] KazusakaA,ShimamotoY,TasakiT,KitamuraHHiroseK: "Decrese in hepatic CYP2C11 mRNA and increase in hemoxygenase activity after intracerbroventricular injection of bacterial endotoxin."J.Vet.Med.Sci. 61. 609-613 (1999)
[Publications] KazusakaA,ShimamotoY,KitamuraH,IwaiM: "Mechanism of decrese in levels of hepatic P450 isozymes induced by intracerebral endotoxin."Arch.Toxicol.. 73. 41-49 (1999)
[Publications] KazusakaA,YamamotoH,HiroseK,HaysakiY,MasudaM: "Mechanism of enhanced lipid peroxidation in the live of Long-Evance cinnamon (LEC) rats."Arch.Toxicol.. 73. 447-464 (1999)
[Publications] KazusakaA,KhloodEBM,MiyoshiH,IwataH,KonY.: "Effects of concurrent exposure to 3-methylcholanthrene and vitamin A on fetal development in rats."Jpn.J.Vet.Res.. 47. 13-23 (1999)
[Publications] KazusakaA,TakahashiJ,FuruhataY,IkedaA,TakahashiM: "Characterization of hepatic cytochrome P450 isozyme composition in the transgenicrat expressing low level human growth hormone."Xenobiotica. 29. 1203-1212 (1999)
[Publications] 数坂昭夫,沈艶波,寺澤実,藤田正一: "シラカンバ樹液の動物生体防御の解明"木材学会誌. 31. 75-78 (1999)
[Publications] KazusakaA,YoonS,FujitaS: "Accumulation of diacylglycerole in the liver membrane of Long-Evance cinnamon (LEC) rat with hepatitis. "Cancer lettr. 151. 19-24 (2000)
[Publications] KazusakaA,YoonS,FujitaS,: "Application of FT-IR and ESR spectroscopic techniques to the study of CC14-induced peroxidation in rat liver microsomes. "Jpn.J.Vet.Res.. 47. 151-154 (2000)
[Publications] KazusakaA,YoonS,FujitaS,: "Accumulation of diacylglycerole induced by CC14-derived radicals in rat liver membrane and its inhibition with radical trapping reagent-FT-IR spectroscopic and HPLC chromatographic observations-"Jpn.J.Vet.Res.. 47. 135-144 (2000)
[Publications] KazusakaA,YoonS,FujitaS,: "FT-IR spectroscopic and HPLC chromatographic studies of CC14-induced acute hepatits in rat-Danmage in lever phospholipid membrane-"Biospictroscopy. 57. 267-271 (2000)
[Publications] KazusakaA,TeramitsuI,YamamotoY,ChidaI.IwataH,TanabeS.: "Identification of novel chtochrome P4501A genens from five marine mannal species."Aquatic Toxicol. 51. 145-153 (2000)
[Publications] KazusakaA,YamamotoH,WatanabeT,MizunoH,EndoK.: "The antioxidant effect of DL-a-linpoic acid on coppe-induced acute hepatitis in Long-Evance cinnamon (LEC) rats."Free Radical Res. 34. 69-80 (2001)
[Publications] KazusakaA,YamamotoH,WatanaeT,MizunoH,EndoK: "In vivo evidence for accelerated generation of hydroxyl radicals in liver of Long-Evance cinnamon (LEC) rats with acute hepatitis."Free Radical Biology and Medicine. 30. 547-554 (2001)
[Publications] KazusakaA,YoonS,FujitaS,: "Infrared spectroscopic study on the mechanism of CC14 toxicity toward membrane phospholipids from rat liver."J.Toxicol.Sci.. 24. 370 (1999)
[Publications] KazusakaA,ChibaI,SakakibaraA,IwataH,TanabeS.: "Seal liber P450, as marker enzyme for marine pollution."J.Toxicol.Sci.. 24. (1999)
[Publications] KazusakaA,MaruyamaY,SuzukiY,TeradaH,IwataH.: "Norsalsolinol, an endogeneous dopaminergic neurotoxin, inhibits dopamine secretion in PC12 cells."J.Toxicol.Sci.. 24. (1999)
[Publications] KazusakaA,YoonS,FujitaS,: "Fourier transform spectroscopy, 12^<th> international conference."Waseda University. 475-476 (1999)
[Publications] KazusakaA,YoonS,FujitaS.: "Generation of diacylglycerol from microsomes during cytochrome P450 mediated hepatotoxicity by carbon mtetrachliride."The 11 th International Conference on Cytochrome P450. 170 (1999)
[Publications] KazusakaA,FujitaS,IwataH,IshizukaM,HoshiH: "Cytochrome P450 in wild animals as a biomarker of environmental pollution."The 11 th International Conference on Cytochrome P450. 170 (1999)
[Publications] KazusakaA,FujitaS,IwataH,IshizukaM,HoshiH: "P450 in wild animals as a biomarker of environmental impact."Scientific Workship on biomarkers in environmental toxicology, Cristchurch, New Zealand. (1999)
[Publications] KazusakaA,ChibaI,Sakakibara,IwataH: "Liver P450 as a biomarker of environmental pollution."2 nd brain seminar cytochrome P450and plant genetic engineering Kobe. (1999)
[Publications] KazusakaA,,TakahashiJ,SakamotoK,MaedaY: "Effect of neonatal exposure to enzo (a) pyrene, endocrine disrupter on P450 expression in rat liver."2 nd brain seminar cytochrome P450 and plant genetic engineering, Kobe. (1999)
[Publications] KazusakaA,,Shen,Y,B,TerasawaM,FujitaS: "Preventive properties of birch sap against oxidative stress in rats."Proceedings of the 2 nd international symposium on sap utilization in bifuka. 149-153 (2000)

2000 Fiscal Year Annual Research Report

生体内酵素反応解析用光ファイバセンサシステムの開発とその発癌機構解明への応用

Principal Investigator

佐々木 一正 北海道工業大学, 工学部, 教授 (00002006)

Research Products

[Publications] 吉田勝仁,長牛英雄,佐々木一正,沈艶波,数坂昭夫 他2名: "生体用光ファイバセンシングシステムの試作と評価"社団法人 電子情報通信学会 技術報告集. OFT2000-8. 47-52 (2000)

[Publications] 吉田勝仁,長牛英雄,佐々木一正,沈艶波,数坂昭夫: "光ファイバセンサ方式による代謝酵素CytochromeP450センシングシステムの開発と特性"北海道工業大学研究紀要 第29号. 第29号. 1-6 (1999)

[Publications] IsseiSasaki,AyakoKawanabe,Isao YamakawaAkio Kazusaka: "In vivo Observation of Cytochrome P4501A Isozyme Reaction in Liver of Living Rats by Using Fiber-Optic Sensor"Conference Proceedings of 13^<th> International Conference on Optical Fiber Sensor. 179-182 (1999)

[Publications] KazusakaA,NakamraA,HirotaT,MorinoA,ImaokaS,FunaeY,YamamotoY: "Cytochrome P-450isoforms responsible for the N-demethylation and cyclohexene-hydroxylation of NS-21"Xenobiotica. 29. 243-252 (1999)

[Publications] KazusakaA,ShimamotoY,TasakiT,KitamuraHHiroseK: "Decrese in hepatic CYP2C11 mRNA and increase in hemoxygenase activity after intracerbroventricular injection of bacterial endotoxin."J.Vet.Med.Sci. 61. 609-613 (1999)

[Publications] KazusakaA,ShimamotoY,KitamuraH,IwaiM: "Mechanism of decrese in levels of hepatic P450 isozymes induced by intracerebral endotoxin."Arch.Toxicol.. 73. 41-49 (1999)

[Publications] KazusakaA,YamamotoH,HiroseK,HaysakiY,MasudaM: "Mechanism of enhanced lipid peroxidation in the live of Long-Evance cinnamon (LEC) rats."Arch.Toxicol.. 73. 447-464 (1999)

[Publications] KazusakaA,KhloodEBM,MiyoshiH,IwataH,KonY.: "Effects of concurrent exposure to 3-methylcholanthrene and vitamin A on fetal development in rats."Jpn.J.Vet.Res.. 47. 13-23 (1999)

[Publications] KazusakaA,TakahashiJ,FuruhataY,IkedaA,TakahashiM: "Characterization of hepatic cytochrome P450 isozyme composition in the transgenicrat expressing low level human growth hormone."Xenobiotica. 29. 1203-1212 (1999)

[Publications] 数坂昭夫,沈艶波,寺澤実,藤田正一: "シラカンバ樹液の動物生体防御の解明"木材学会誌. 31. 75-78 (1999)

[Publications] KazusakaA,YoonS,FujitaS: "Accumulation of diacylglycerole in the liver membrane of Long-Evance cinnamon (LEC) rat with hepatitis. "Cancer lettr. 151. 19-24 (2000)

[Publications] KazusakaA,YoonS,FujitaS,: "Application of FT-IR and ESR spectroscopic techniques to the study of CC14-induced peroxidation in rat liver microsomes. "Jpn.J.Vet.Res.. 47. 151-154 (2000)

[Publications] KazusakaA,YoonS,FujitaS,: "Accumulation of diacylglycerole induced by CC14-derived radicals in rat liver membrane and its inhibition with radical trapping reagent-FT-IR spectroscopic and HPLC chromatographic observations-"Jpn.J.Vet.Res.. 47. 135-144 (2000)

[Publications] KazusakaA,YoonS,FujitaS,: "FT-IR spectroscopic and HPLC chromatographic studies of CC14-induced acute hepatits in rat-Danmage in lever phospholipid membrane-"Biospictroscopy. 57. 267-271 (2000)

[Publications] KazusakaA,TeramitsuI,YamamotoY,ChidaI.IwataH,TanabeS.: "Identification of novel chtochrome P4501A genens from five marine mannal species."Aquatic Toxicol. 51. 145-153 (2000)

[Publications] KazusakaA,YamamotoH,WatanabeT,MizunoH,EndoK.: "The antioxidant effect of DL-a-linpoic acid on coppe-induced acute hepatitis in Long-Evance cinnamon (LEC) rats."Free Radical Res. 34. 69-80 (2001)

[Publications] KazusakaA,YamamotoH,WatanaeT,MizunoH,EndoK: "In vivo evidence for accelerated generation of hydroxyl radicals in liver of Long-Evance cinnamon (LEC) rats with acute hepatitis."Free Radical Biology and Medicine. 30. 547-554 (2001)

[Publications] KazusakaA,YoonS,FujitaS,: "Infrared spectroscopic study on the mechanism of CC14 toxicity toward membrane phospholipids from rat liver."J.Toxicol.Sci.. 24. 370 (1999)

[Publications] KazusakaA,ChibaI,SakakibaraA,IwataH,TanabeS.: "Seal liber P450, as marker enzyme for marine pollution."J.Toxicol.Sci.. 24. (1999)

[Publications] KazusakaA,MaruyamaY,SuzukiY,TeradaH,IwataH.: "Norsalsolinol, an endogeneous dopaminergic neurotoxin, inhibits dopamine secretion in PC12 cells."J.Toxicol.Sci.. 24. (1999)

[Publications] KazusakaA,YoonS,FujitaS,: "Fourier transform spectroscopy, 12^<th> international conference."Waseda University. 475-476 (1999)

[Publications] KazusakaA,YoonS,FujitaS.: "Generation of diacylglycerol from microsomes during cytochrome P450 mediated hepatotoxicity by carbon mtetrachliride."The 11 th International Conference on Cytochrome P450. 170 (1999)

[Publications] KazusakaA,FujitaS,IwataH,IshizukaM,HoshiH: "Cytochrome P450 in wild animals as a biomarker of environmental pollution."The 11 th International Conference on Cytochrome P450. 170 (1999)

[Publications] KazusakaA,FujitaS,IwataH,IshizukaM,HoshiH: "P450 in wild animals as a biomarker of environmental impact."Scientific Workship on biomarkers in environmental toxicology, Cristchurch, New Zealand. (1999)

[Publications] KazusakaA,ChibaI,Sakakibara,IwataH: "Liver P450 as a biomarker of environmental pollution."2 nd brain seminar cytochrome P450and plant genetic engineering Kobe. (1999)

[Publications] KazusakaA,,TakahashiJ,SakamotoK,MaedaY: "Effect of neonatal exposure to enzo (a) pyrene, endocrine disrupter on P450 expression in rat liver."2 nd brain seminar cytochrome P450 and plant genetic engineering, Kobe. (1999)

[Publications] KazusakaA,,Shen,Y,B,TerasawaM,FujitaS: "Preventive properties of birch sap against oxidative stress in rats."Proceedings of the 2 nd international symposium on sap utilization in bifuka. 149-153 (2000)

佐々木一正北海道工業大学, 工学部, 教授 (00002006)

[Publications] 吉田勝仁,長牛英雄,佐々木一正,沈艶波,数坂昭夫他2名: "生体用光ファイバセンシングシステムの試作と評価"社団法人電子情報通信学会技術報告集. OFT2000-8. 47-52 (2000)

[Publications] 吉田勝仁,長牛英雄,佐々木一正,沈艶波,数坂昭夫: "光ファイバセンサ方式による代謝酵素CytochromeP450センシングシステムの開発と特性"北海道工業大学研究紀要第29号. 第29号. 1-6 (1999)