Relationship between xenobiotic toxicity and metabolisn in individuals

异生物质毒性与个体代谢的关系

• 批准号: 18590116
• 负责人: HANIOKA Nobumitsu
• 金额: $ 2.53万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590116/
• 关键词: Environmental pollutants; Drug-metabolizing enzymes; UDP-glucuronosyltransferase; β-nanhthoflavone; HepG2 cells; UGT1A1; UGT1A6; UGTIA9; シトクロムP450(CYP); UDP-グルクロン酸転移酵素(UGT); A549細胞; フェノバルビタール; デキサメタゾン; UGT1A1; UGT1A9

Since glucuronidation is an important metabolic reaction for xenobiotic elimination, information on the induction or suppression of UDP-glucuronosyltransferase (UGT) enzymes at the isoform level is beneficial in drug therapy and the toxicological assessment of environmental chemicals. In this study, we focused on UGT1A isoforms (UGT1A1, UGT1A6 and UGT1A9), mainly expressed in the human liver, and examined the inducibility of UGT1As by β-naphthoflavone (BNF) in human hepatoma HepG2 cells. The cells were pretreated for 72 h with BNF at concentrations of 25, 50 and 100 μM. 7-Ethyl-10-hydroxycamptothecin (SN-38) glucuronidation, used as a probe for UGT1A1, showed sigmoidal kinetics with a Hill coefficient (n) of 1.2-1.3 in control and BNF-pretreated HepG2 cells. The V_<max> values were significantly increased 3.6- to 4.3-fold by BNF, whereas there was no significant change in the S_<50> values by BNF at any concentration examined. On the other hand, 4-methylumbelliferone (4-MU) glucuronida … More tion as a probe for UGT1A6 and UGT1A9 in the control and BNF-pretreated HepG2 cells exhibited a biphasic kinetic pattern Although K_<ml> values for the low-K_m phase were similar between the control and BNF-pretreated HepG2 cells, K_<m2> values for the high-K_m phase of BNF-pretreated liepG2 cells were reduced to 54-69% of control HepG2 cells. The values of V_<max> and V_<max2> for the low- and high-K_m phases, respectively, were significantly increased 1.9- to 2.6-fold by BNF at 25 and/or 50μM but not 100 μM With respect to V_<max> (V_<max1> and V_<max2>) and V_<max>/K_m (V_<max1>/K_<m1> and V_<max2>/K_<m2>), the values were significantly increased 2.0- to 3.2-fold by BNF at all concentrations examined. Furthermore, real-time reverse transcription polymerase chain reaction (RT-PCR) using TaqMan probes demonstrated that BNF concentration-dependently induced mRNA levels of UGTIA1 but not UGT1A6 or UGT1A9 in HepG2 cells (1.3- to 6.0-fold). These results suggest that the inducibility of UGT1A isoforms in HepG2 cells by BNF is different from other aryl hydrocarbon receptor (AhR) agonists previously reported, and should provide useful information for the prediction of drug-drug interactions and toxicological assessment of environmental chemicals. Less

由于葡萄糖醛酸化作用是消除异物的重要代谢反应，在异构体水平上诱导或抑制UDP-葡萄糖醛酸基转移酶(UGT)的信息有助于药物治疗和环境化学品的毒理学评估。在本研究中，我们以主要在人肝脏表达的UGT1A1A1、UGT1A6和UGT1A9亚型为研究对象，研究了β-萘黄酮对人肝癌细胞株Hep G2细胞UGT1As的诱导作用。用25、50和100μ的7-乙基-10-羟基喜树碱(SN-38)葡萄糖醛酸化作为UGT1A1的探针，处理细胞72 h后，对照组和处理组细胞的希尔系数(N)为1.2~1.3。在所考察的任何浓度下，巴斯夫能使V&lt；max&gt；值显著增加3.6倍至4.3倍，而对S的值无显著影响。另一方面，4-甲基伞形酮(4-MU)葡萄糖醛酸苷…作为UGT1A6和UGT1A9的探针，对照组和BNF处理的HepG2细胞呈现双时相动力学模式，但低K_m期的K_m；m1和gt；值与对照组相似，高K_m期的K_m值仅为对照组的54%-69%。在25μM和/或50μM时，低K_m相和高K_m相的V_&lt；max&gt；和V_&lt；max&gt；和V_&lt；max&gt；/K_m(V_&lt；max 1&gt；/K_&lt；M1&gt；)和V_&lt；max&gt；/K_m(V_&lt；max 1&gt；/K_&lt；M1&gt；)和V_&lt；max&gt；和V&lt；Max2&gt；/K&lt；m2&gt；)，在所考察的所有浓度下，BNF值均显著增加2.0-3.2倍。此外，利用TaqMan探针的实时逆转录聚合酶链式反应(RT-PCR)显示，BNF浓度依赖地诱导HepG2细胞UGTIA1的mRNA水平，但不能诱导UGT1A6或UGT1A9的mRNA水平(1.3-6.0倍)。这些结果表明，BNF对HepG2细胞UGT1A亚型的诱导作用不同于以往报道的芳烃受体(AhR)激动剂，为药物-药物相互作用的预测和环境化学品的毒理学评估提供了有用的信息。较少

项目成果

Influence of CYP2C19*18 and CYP2C19*19 alleles on omeprazole 5-hydroxylation: in vitro functional analysis of recombinant enzymes expressed in Saccharomyces cerevisiae

CYP2C19 * 18和CYP2C19 * 19等位基因对奥美拉唑5-羟基化的影响：酿酒酵母中表达的重组酶的体外功能分析

• 发表时间: 2008
• 期刊: Basic Clin. Pharmacol. Toxicol. (印刷中)
• 作者: Hanioka;N.;Tsuneto;Y.;Saito;Y.;Maekawa;K.;Sawada;J.;and Narimatsu;S.
• 通讯作者: S.

Effect of SHP on transcriptional activities mediated by human HNF4α

SHP 对人 HNF4α 介导的转录活性的影响

• 发表时间: 2007
• 作者: Nakada;S.;Saito;Y.;Saeki;M.;Sawada;J.;Hanioka;N.;Narimatsu;S
• 通讯作者: S

ヒトHNF4αを介した転写活性化に及ぼすSHPの影響

SHP 对人 HNF4α 介导的转录激活的影响

• 发表时间: 2007
• 作者: 中田晋太郎;斎藤嘉朗;佐伯真弓;澤田純一;埴岡伸光;成松鎭雄
• 通讯作者: 成松鎭雄

Expression and inducibility of drug-metabolizing enzymes in MCF-7 cells

MCF-7细胞中药物代谢酶的表达和诱导能力

• 发表时间: 2007
• 作者: Iwabu;H.;Hanioka;N.;Narimatsu;S
• 通讯作者: S

Functional characterization of human and cynomolgus monkey cytochrome P450 2Elenzymes

人和食蟹猴细胞色素 P450 2Elenzymes 的功能表征

• 发表时间: 2007
• 期刊: Life Sci. 81(19-20)
• 作者: Hanioka;N.;Yamamoto;M.;Iwabu;H.;Jinno;H.;Tanaka-Kagawa;T.;Naito;S.;Shimizu;T.;Masuda;K.;Katsu;T.;and Narimatsu;S.
• 通讯作者: S.