Evaluation of reliability of human liver surgical samples from hepatocellular carcinoma and metastatic cancer patients on the application to the drug-metabolism study

肝细胞癌和转移癌患者肝脏手术样本应用于药物代谢研究的可靠性评价

• 批准号: 11672277
• 负责人: KURATA Norimitsu
• 金额: $ 2.3万
• 依托单位: SHOWA University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11672277/
• 关键词: Human liver; Surgical Samples; Drug metabolism; Cytochrome P-450; microsomes; CYP; ヒト肝臓; cytochrome P450; 手術材料; 薬物代謝

Reliability of human surgical liver sample from hepatocellular carcinoma (HCC) patients and metasatic hepatic cancer (MHC) patients on the study of drug metabolism has been evaluated. Liver samples were donated from 7 HCC and 4 MHC patients. Liver sample was divided into three parts, carcinoma, peripheral parts (within 10 mm from carcinoma) and normal parts (10 mm or more further from carcinoma). Microsomal fraction in each parts, carcinoma, peripheral parts and normal parts, were prepared by conventional method. Human pooled hepatic microsome from Caucasian brain death patients was employed as a reference enzyme source. Microsomal Cytochrome P450 (CYP) enzyme activity, CYP1A2 (Caffeine, Ethoxyresorfin), 2C9 (Warfarin), 2C19 (Mephenytoin), 2D6 (Bufuralol), 2E1 (Chlorzoxazone) and 3A4 (Midazolam, Testosterone), were analyzed by using typical substrates. HPLC method was employed for the determination of each enzyme activity. CYP enzymes activity in normal parts was variated from 2 to 5 fold. Difference of CYP activity between normal parts and peripheral parts were within 2 fold in all liver samples. Furthermore, mean activity in 7 liver samples in each CYP enzymes and pooled microsome were quite similar in most of CYP enzymes. In CYP2C19, however, enzyme activity was quite low compared with pooled microsome. In conclusion, human surgical liver samples will be one of the enzyme sources for the estimation of drug metabolism in human. Further study will be needed on the lower activity on CYP 2C19 from surgical liver samples.

本文评价了肝细胞癌（HCC）和转移性肝癌（MHC）患者手术肝标本用于药物代谢研究的可靠性。肝脏样本来自7名HCC和4名MHC患者。将肝脏样品分为三个部分，癌、周围部分（距癌10 mm内）和正常部分（距癌10 mm或更远）。用常规方法制备癌组织、外周组织和正常组织的微粒体组分。采用来自高加索脑死亡患者的人混合肝微粒体作为参考酶源。使用典型底物分析微粒体细胞色素P450（CYP）酶活性、CYP 1A 2（咖啡因、乙氧基间苯二酚）、2C 9（华法林）、2C 19（美芬妥英）、2D 6（丁呋洛尔）、2 E1（氯唑沙宗）和3A 4（咪达唑仑、替吉奥）。采用高效液相色谱法测定各酶的活性。正常组织中的酶活性变化为2 ~ 5倍。所有肝组织中，正常肝组织与周围肝组织的γ-淀粉酶活性差异均在2倍以内。此外，7份肝脏样品中每种酶和混合微粒体的平均活性在大多数酶中非常相似。然而，在CYP 2C 19中，与合并微粒体相比，酶活性相当低。结论：人手术肝标本可作为人体药物代谢的酶源之一。对手术肝样品中较低的B_2C19活性需要进一步研究。

项目成果

Norimitsu Kurata: "Evaluation of reliability of human liver surgical sample for the application to the study of estimation of drug-metabolizing enzyme capacity"The Japanese J.Pharmacology. Suppl.1. 101p (2000)

Norimitsu Kurata：“评估人类肝脏手术样本的可靠性，用于估计药物代谢酶能力的研究”，日本药理学杂志。

Norimitsu Kurata, Mariko Iwase, Hua Li, Yuki Nishimura, Youji Ito, Mitsuo Kusano, Eiji Uchida and Hajime Yasuhara: "Reliability of human liver surgical waste donated from cancer patients for the study of drug metabolism."The J.Pharmacology. Suppl.1. 183 (

Norimitsu Kurata、Mariko Iwase、Hua Li、Yuki Nishimura、Youji Ito、Mitsuo Kusano、Eiji Uchida 和 Hajime Yasuhara：“癌症患者捐赠的人类肝脏手术废物用于药物代谢研究的可靠性。”J.Pharmacology。

倉田知光: "薬物代謝研究におけるヒト肝手術材料の有用性に関する検討"臨床薬理. 31. 271-272 (2000)

Tomomitsu Kurata：“人体肝脏手术材料在药物代谢研究中的有用性研究”临床药理学31。271-272（2000）。

倉田知光、西村有希、岩瀬万里子、李華、植田孝子、内田英二、安原 一: "薬物代謝研究におけるヒト肝手術材料の有用性に関する検討"臨床薬理. 31(2)(in press). (2000)

Tomomitsu Kurata，Yuki Nishimura，Mariko Iwase，Li Hua，Takako Ueda，Eiji Uchida，Hajime Yasuhara：“人体肝脏手术材料在药物代谢研究中的有用性研究”临床药理学31（2）（出版中）。

Yuki Nishimura, Norimitsu Kurata, Imad H.Hanna, F.Peter Guengerich and Hajime Yasuhara: "Stability of cytochrome P450 enzymes in human liver samples stored in different tissue preparation buffers"Xenobiotica. (Submitted).

Yuki Nishimura、Norimitsu Kurata、Imad H.Hanna、F.Peter Guengerich 和 Hajime Yasuhara：“保存在不同组织制备缓冲液中的人类肝脏样品中细胞色素 P450 酶的稳定性”Xenobiotica。