Development of monitoring system of toxic chemicals base on the Nrf2-dependent detoxification pathway

基于Nrf2依赖性解毒途径的有毒化学品监测系统的开发

• 批准号: 12557015
• 负责人: YAMAMOTO Masayuki
• 金额: $ 8.58万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557015/
• 关键词: electrophiles; xenobiotics; transcriptional regulation; chemicals; Nrf2; oxidative stress; mouse; zebrafish; 環境異物; モデル動物; bZip型転写因子; bHLH-PAS型転写因子

Analysis using Nrf2-LacZ miceAfter feeding NrfZ-LacZ mice with dietary BHA for 3 days, accumulation of LacZ activity was observed in intestine, liver and peritoneal macrophages. The result suggested that Nrf2-LacZ mice are useful for detecting toxic chemicals in foods or medicines. In addition, we found a posttranseriptional accumulation of Nrf2 protein in response to electrophiles.Analysis of Nrf2 knockout miceNrf2 knockout mice was susceptible to benzo[o]pyrene-induced neoplasia in the forestomach, diesel exhaust-induced hyperplasia and oxidative DNA adduct formation in the lung, and acetaminophen-induced hepatotoxicity. These results demonstrated that Nrf2 knockout mice are highly sensitive to carcinogen and/or oxidative stress, and that they will be useful for detecting toxic chemicals in foods or medicines.Zebrafish system for xenobiotic studiesWe showed that Nrf2-based induction of phase II detoxifying enzymes was conserved in fish and that a possibility of monitoring carcinogens using zebra fish system. Thus the GFP reporter construct fused to the transcriptional regulatory region of zebrafish GSTpi gene was prepared. In a transient reporter analysis, we could observed a GFP induction by electrophiles in zebra fish larvae. We are now trying to establish a stable transgenic zebrafish line which contains the construct.

使用 Nrf2-LacZ 小鼠进行分析 用膳食 BHA 喂养 NrfZ-LacZ 小鼠 3 天后，在肠道、肝脏和腹膜巨噬细胞中观察到 LacZ 活性的积累。结果表明，Nrf2-LacZ 小鼠可用于检测食品或药品中的有毒化学物质。此外，我们发现 Nrf2 蛋白在对亲电子试剂的反应中出现转录后积累。 Nrf2 敲除小鼠的分析 Nrf2 敲除小鼠易受苯并[o]芘诱导的前胃肿瘤、柴油机尾气诱导的增生和氧化 DNA 加合物形成的影响。肺和对乙酰氨基酚引起的肝毒性。这些结果表明，Nrf2 敲除小鼠对致癌物和/或氧化应激高度敏感，并且它们可用于检测食品或药物中的有毒化学物质。用于异生素研究的斑马鱼系统我们表明，基于 Nrf2 的 II 相解毒酶诱导是保存在鱼类中，并且可以使用斑马鱼系统监测致癌物。由此制备了与斑马鱼GSTpi基因转录调控区融合的GFP报告基因构建体。在瞬时报告分析中，我们可以观察到斑马鱼幼虫中亲电子试剂对 GFP 的诱导。我们现在正在尝试建立包含该构建体的稳定的转基因斑马鱼品系。

项目成果

Hoshino,H.: "Oxidative stress abolishes leptomycine B-sensitive nuclear export of transcription repressor Bach2 that counteracts activation of Maf recognition element."J.Biol.Chem.. 275. 15370-15376 (2000)

Hoshino, H.：“氧化应激消除了转录抑制子 Bach2 的细霉素 B 敏感核输出，从而抵消了 Maf 识别元件的激活。”J.Biol.Chem.. 275. 15370-15376 (2000)

Sun, J.: "The promoter of mouse transcription repressor bach1 is regulated by Sp1 and transactivated by Bach1"J. Biochem.. 130. 385-392 (2001)

Sun, J.：“小鼠转录抑制子 bach1 的启动子受 Sp1 调节并被 Bach1 反式激活”。

鈴木教郎, 山本雅之: "造血幹細胞:基礎から遺伝子治療・再生医療へ 「造血幹細胞の転写調節機構」"中外医学社(印刷中).

Norio Suzuki、Masayuki Yamamoto：“造血干细胞：从基础知识到基因治疗和再生医学‘造血干细胞的转录调控机制’”中外医学社（出版中）。

Katoh, Y, Itoh, K., Yoshida, E., Miyagishi, M., Fukamizu, A., Yamamoto, M.: "Two domains of Nrf2 cooperatively bind CBP (CREB binding protein) and ynergistically activate transcription"Genes Cells. 6. 857-868 (2001)

Katoh, Y、Itoh, K.、Yoshida, E.、Miyagishi, M.、Fukamizu, A.、Yamamoto, M.：“Nrf2 的两个结构域协同结合 CBP（CREB ​​结合蛋白）并协同激活转录”Genes Cells。

Hayes, J.D., Chanas, S.A., Henderson, C.J., McMahon, M., Sun, C., Moifat, G.J., Wolf, C.R., Yamamoto, M.: "The Nrf2 transcription factor contributes both to the basal expression of glutathione S-transferases in mouse liver and to their induction by the ch

Hayes, J.D.、Chanas, S.A.、Henderson, C.J.、McMahon, M.、Sun, C.、Moifat, G.J.、Wolf, C.R.、Yamamoto, M.：“Nrf2 转录因子有助于谷胱甘肽 S- 的基础表达