Development of multi-generation assay for endocrine disrupting chemicals

内分泌干​​扰物多代检测方法的开发

• 批准号: 13833009
• 负责人: TOMINAGA Nobuaki
• 金额: $ 2.37万
• 依托单位: Ariake National College of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13833009/
• 关键词: endocrine disruptor; C.elegans; bioassay; reproduction; multi-generation; 内分泌攪乱物質; ビスフェノールA; 環境ホルモン; エストロジェン; アンドロジェン

In this study, we investigated to develop a multiple-generation toxicity assay for biological evaluation of chemicals. The nematode Caenorhabditis elegans was adopted for a model organism. Then we examined the sublethal toxicity, fecundity and reproduction for over five generations. As the results, C. elegans responds to vertebrate steroid hormones. Especially estradiol has a cholesterol-like potency in supporting the reproduction of C. elegans. In contrast testosterone and diethylstibestrol suppressed the fecundity rate of C. elegans. Some chemicals were evaluated by same method. In particular, bisphenol A was decreased the number of C. elegans significantly even at 1 nM. A comparison of the number of viable worms and eggs suggested that the phenols exert hatching toxicity.When we performed western-blotting analyses using specific antibodies for phosphorylated amino acids, we could detect some proteins which phosphorylation levels were changed in estradiol exposed C. elegans extracts.These results indicated that this toxicity assay had 100-1000-fold higher sensitivity than the LC50s and free living nematode C. elegans is becoming an effective model organism for testing chemical toxicity including reproductive disruption.

在这项研究中，我们研究开发了一种用于化学品生物学评价的多代毒性测定方法。采用线虫秀丽隐杆线虫作为模式生物。然后我们检查了五代以上的亚致死毒性、繁殖力和繁殖能力。结果，线虫对脊椎动物类固醇激素产生反应。尤其是雌二醇具有类似胆固醇的功效，可以支持线虫的繁殖。相反，睾酮和二乙基己雌酚抑制了秀丽隐杆线虫的繁殖率。一些化学物质也用同样的方法进行了评价。特别是，双酚 A 即使在 1 nM 的浓度下也会显着减少线虫的数量。活虫和卵数量的比较表明酚类具有孵化毒性。当我们使用磷酸化氨基酸的特异性抗体进行蛋白质印迹分析时，我们可以检测到雌二醇暴露的秀丽隐杆线虫提取物中一些磷酸化水平发生变化的蛋白质这些结果表明，该毒性测定的灵敏度比 LC50 高 100-1000 倍，并且自由生活的线虫秀丽隐杆线虫正在成为测试化学物质的有效模型生物毒性包括生殖障碍。

项目成果

Shinya Kohra, Nobuaki Tominaga, Yuji Takao, Masaki Nagae, Yasuhiro Ishibashi, Kazuo Ueda, Koji Arizono: "A rapid respiratory toxicity test using Caenorhabditis elegans with an oxygen electrode system"J.Health Science. 48(3). 269-272 (2002)

Shinya Kohra、Nobuaki Tominaga、Yuji Takao、Masaki Nagae、Yasuhiro Ishibashi、Kazuo Ueda、Koji Arizon：“使用带有氧电极系统的秀丽隐杆线虫进行快速呼吸道毒性测试”J.Health Science。

Tetsuya Matsuno, Kazuhiro Ura, Risa Sonoda, Yuji Kohara, Hiroko Uesugi, Koji Arizono, Taisen Iguchi, Nobuaki Tominaga: "Sensing of chemical substances using gene expression patterns in C. elegans"Sensors & Materials. 14(7). 395-406 (2002)

Tetsuya Matsuno、Kazuhiro Ura、Risa Sonoda、Yuji Kohara、Hiroko Uesugi、Koji Arizono、Taisen Iguchi、Nobuaki Tominaga：“利用线虫中的基因表达模式感知化学物质”传感器

Shinya Kohra, Nobuaki Tominaga, Yuji Takao, Masaki Nagae, Yasuhiro Ishibashi, Kazuo Ueda, Kqji Arizono: "A rapid respiratory toxicity test using Caenorhabditis elegans with an oxygen electrode system"Journal of Health Science. 48(3). 262-272 (2002)

Shinya Kohra、Nobuaki Tominaga、Yuji Takao、Masaki Nagae、Yasuhiro Ishibashi、Kazuo Ueda、Kqji Arizono：“使用带有氧电极系统的秀丽隐杆线虫进行快速呼吸道毒性测试”健康科学杂志。

S.Kohra et al.: "Effect of BisphenolA on the feeding behavior of Caenorhabditis elegans"Journal of Health Science. 48・1. 93-95 (2002)

S. Kohra 等人：“双酚 A 对秀丽隐杆线虫摄食行为的影响”《健康科学杂志》48・1（2002 年）。

Nobuaki Tominaga, Mineko Tomoeda, Shinya Kohra, Yuji Takao, Masaki Nagae, Kazuo Ueda, Yasuhiro Ishibashi, Toshinori Kai, Koji Arizono: "A convienient sublethal assay of alkylphenol and organotin compounds using the nematode Caenorhabditis elegans"Journal

Nobuaki Tominaga、Mineko Tomoeda、Shinya Kohra、Yuji Takao、Masaki Nagae、Kazuo Ueda、Yasuhiro Ishibashi、Toshinori Kai、Koji Arizono：“使用线虫秀丽隐杆线虫对烷基酚和有机锡化合物进行便捷的亚致死测定”期刊