Effects of fetal and neonatal exposure of male mice to diethylstilbestrol as endocrine disrupter on the reproductive and neuroendocrine system

雄性小鼠胎儿和新生儿暴露于己烯雌酚作为内分泌干扰物对生殖和神经内分泌系统的影响

• 批准号: 12836014
• 负责人: HOSHI Nobuhiko
• 金额: $ 2.05万
• 依托单位: Kitasato University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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/grant/KAKENHI-PROJECT-12836014/
• 关键词: Endcrine disrupter chemicals; Sex determination; Sex differentiation; diethylstilbestrol; spermatogenesis; mouse; hypothalamus- hypophysis-gonadal axis; 内分泌撹乱化学物質; 脳-下垂体-性腺系

The purpose of this study is to determine the effect of fetal and neonatal exposure to diethylstilbestrol (DBS), as a representative endocrine disrupter, on the reproductive function of male mice. Various doses (0.001-100 μg) of DES or a vehicle (control) were administered to the neonatal group on day 3 (single) and on every 2 days from day 3 (N-long-term), and to the fetal group every 2 days from 7 dpc (F-long-term). Endocrinological and morphological changes were evaluated at 25 days of age (immediately before the onset of puberty), 4, 6, 8, and 12 weeks of age. We established a non-RI solid phase immunoassay method, which enables multiple microassay (LH, FSH, PRL, testosterone, inhibin) of the same sample with high sensitivity and specificity. In single administration of 10 and 100 μg of DES at 4 weeks, N-long-term administration at 1.0 μg for 25 days and 10 μg for 6 weeks, aid F-long-term administration at 10 μg for 6 weeks, testis weights were significantly reduced, and no spermat … More ogenesis and immature seminiferous tubules were observed. DES injections caused dose-dependent retardation of pituitary LH, FSH. Using the morphometric immunodetection method for incorporated thymidine analog BrdU, the high-dose groups (10 ＆ 100 μg) showed a marked disruption of sperm production and significant suppression of cellular proliferation in testes. Immunohistochemical analysis in each group showed that immunoreactive neurons for GnRH were generally fewer in number and had thinner neurites in the high-dose groups Immunoreactivity of Era and StAR (steroidogenic acute regulatory protein) in testes was decreased. Thus our findings indicate that 1) single neonatal exposure of male mice to 10 μg of DES can advance the spermatogenesis at 6 weeks of age; 2) long-term exposure of these mice, particularly from fetus, could not recover the spermatogenesis even under 1.0 μg of DES; and 3) there is threshold dose of DES, and these delay and damage in the reproduction system due to reduction of pituitary gonadotropin production. Less

本研究的目的是确定胎儿和新生儿接触内分泌干扰物己烯雌酚（DBS）对雄性小鼠生殖功能的影响。新生儿组在第3天（单天）和第3天（n -长期）每2天给药一次（0.001 ~ 100 μg），胎儿组从第7天开始每2天给药一次（f -长期）。在25日龄（青春期开始前）、4、6、8和12周龄时评估内分泌和形态学变化。我们建立了一种非ri固相免疫分析方法，可以对同一样品进行多种微量分析（LH、FSH、PRL、睾酮、抑制素），具有较高的灵敏度和特异性。单次给药10和100 μg（4周）、n -长期给药1.0 μg（25天）和10 μg（6周）、f -长期给药10 μg（6周）各组小鼠睾丸重量显著降低，无精子发生，精子小管发育不成熟。DES注射剂对垂体LH、FSH的抑制呈剂量依赖性。结合胸腺嘧啶类似物BrdU的形态免疫检测方法显示，高剂量组（10和100 μg）明显破坏精子产生并显著抑制睾丸细胞增殖。各组免疫组化分析显示，高剂量组GnRH免疫反应神经元数量普遍减少，神经突变细。结果表明：1)雄鼠单次暴露于10 μg DES可促进6周龄时的精子发生；2)长期暴露于小鼠，尤其是从胎儿处，即使在1.0 μg的DES浓度下，也不能恢复小鼠的精子发生能力；(3) DES存在阈值剂量，并且由于垂体促性腺激素的产生减少而延迟和损害生殖系统。少

项目成果

Sugawara T et al.: "Sp1 and SF-1 interact and cooperate in the regulation of human steroidogenic acute regulatory protein gene expression"Endocrinology. 141. 2895-2903 (2000)

Sukawara T 等人：“Sp1 和 SF-1 在人类类固醇生成急性调节蛋白基因表达的调节中相互作用和配合”内分泌学。

Yamamoto R: "Lens culinaris agglutinin-reactive α-fetoprotein, an alternative variant to α-fetoprotein in prenatal screening for Down's syndrome"Hum Reprod. 16・11. 2438-2444 (2001)

Yamamoto R：“Lens culinaris 凝集素反应性 α-胎蛋白，唐氏综合症产前筛查中的 α-胎蛋白的替代变体”Hum Reprod 16・11 (2001)。

Sugawara T: "The effect of the aryihydrocarbon receptor on the human steroidogenie acute regulatory gene promoter activity"Steroid Biochem Mol Biol. 78. 253-260 (2001)

Sukawara T：“芳基烃受体对人类类固醇生成急性调节基因启动子活性的影响”Steroid Biochem Mol Biol。

Yamamoto R et al.: "Lens culinaris agglutinin-reactive α-fetoprotein, an alternative variant to a-fetoprotein in prenatal screening for Down's syndrome"Hum Reprod. 16. 2438-2444 (2001)

Yamamoto R 等人：“Lens culinaris 凝集素反应性 α-胎蛋白，唐氏综合症产前筛查中甲胎蛋白的替代变体”Hum Reprod. 16. 2438-2444 (2001)

Sugawara T et al.: "RIP 140 modulates transcription of the steroidogenic acute regulatory protein gene through interactions with both SF-1 and DAX-1"Endocrinology. 142. 3570-3577 (2001)

Sukawara T 等人：“RIP 140 通过与 SF-1 和 DAX-1 的相互作用调节类固醇生成急性调节蛋白基因的转录”内分泌学。