邻苯二甲酸二丁酯对亲代和子代斑马鱼生殖毒性的比较研究

doi:10.3969/j.issn.1674-5817.2012.05.003

实验动物与比较医学 ›› 2012, Vol. 32 ›› Issue (5): 383-389.DOI: 10.3969/j.issn.1674-5817.2012.05.003

• 华东地区第12 届实验动物科学学术交流会优秀论文选刊 • 上一篇下一篇

邻苯二甲酸二丁酯对亲代和子代斑马鱼生殖毒性的比较研究

徐浩¹, 王玉柱², 温成丽², 成倩倩², 田芳², 夏敏杰², 李润生², 李卫华², 丁训诚²

1.上海中医药大学附属龙华医院, 200032 上海;
2.上海市计划生育科学研究所, 200032 上海

收稿日期:2012-02-06 出版日期:2012-10-25 发布日期:2012-10-25
作者简介:徐浩(1981-),男,博士研究生,从事中医骨伤科学研究,E-mail:hoxu@163.com
基金资助:
上海市科学技术委员会(09140903200)

Reproductive Toxicity of Dibutyl Phthalate in Parent and Progeny Zebrafish

XU Hao¹, WANG Yu-zhu², WEN Cheng-li², CHENG Qian-qian², TIAN Fang², XIA Min-jie², LI Run-sheng², LI Wei-hua², DING Xun-cheng²

1. Longhua Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China;
2. Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China

Received:2012-02-06 Online:2012-10-25 Published:2012-10-25

摘要/Abstract

摘要： 目的探讨邻苯二甲酸二丁酯(DBP)对亲代(F₀)、子代(F₁)斑马鱼的生殖毒性。方法将48对成年斑马鱼随机分为4组: 空白对照组、溶剂对照组、625 µg/L DBP低剂量组和1 250 µg/L DBP高剂量组。连续染毒DBP 30 d后,将各组F₀代斑马鱼交配,记录产卵数、受精率以及72 h孵化率。将F₀和F₁代斑马鱼在清水中饲养180 d,进行F₁代斑马鱼交配试验,且将F₁代DBP高剂量组与对照组斑马鱼进行置换交配,记录产卵数、受精率及孵化率。进行F₀代和F₁代斑马鱼性腺组织病理学检查,并测定卵黄蛋白原(VTG) mRNA的表达水平。结果与对照组和DBP低剂量组相比,DBP高剂量组F₀代和F₁代斑马鱼的生殖能力显著下降;置换交配实验显示DBP对雄性斑马鱼的生殖能力影响更为显著。组织病理学检查显示F₀代雄鱼精巢中精子数量减少,间质细胞稍有增多,精母细胞增多。而F₁代雄鱼精巢发育受到明显抑制,出现生精小管畸形,精子数量明显减少,以及间质细胞增生。RT-PCR结果显示DBP各剂量组F₀代和F₁代雄性斑马鱼VTG基因无明显表达,提示DBP对斑马鱼无雌激素样效应。结论 DBP高剂量组染毒可影响F₀代和F₁代斑马鱼的正常发育,使产卵量减少,受精率显著下降。F₀代和F₁代雄性斑马鱼生精小管畸形、精巢中精子数量减少与间质细胞增生同时存在。

关键词: 斑马鱼, 邻苯二甲酸二丁酯(DBP), 生殖毒性, F₀, F₁

Abstract: Objective To study the reproductive toxicity effects of dibutyl phthalate (DBP) on F₀ (parent) and F₁(parent) zebrafish. Methods Fourty-eight pairs of adult zebrafish were randomly divided into 4 groups: blank control group, 0.01% acetone solvent control group, 625 µg/L DBP low-dose group, and 1250 µg/L DBP high-dose group. After 30-day exposure, F₀ zebrafish in each group were mated and the number of eggs, fertilization rate and 72-hours hatchability were recorded. After being kept for 180 days in clean water, F₀ and F₁ zebrafish in each group were again mated, and the replacement mating test were conducted. The Vetellogenin (VTG) mRNA of F₀ and F₁ zebrafish was measured by RT-PCT, and gonad histopathology was performed. Results The fertilization of F₀ and F₁ zebrafish in 1250 µg/L DBP group was significantly lower than that in blank control or in 625 µg/L DBP group; histological examination showed that DBP slightly inhibited ovarian development of the F₀ and F₁ zebrafish and slightly increased the cortical vesicular stage oocyte (Coc) and perinucleolar stage oocyte (Poc). In F₀ male zebrafish of 1250 µg/L DBP group, the sperm count in the testes was reduced, and the spermatocyte (sc) and Leydig cells were slightly increased. In F₁ male zebrafish of 1250 µg/L DBP group, the development of testes was inhibited significantly, manifested as the co-existence of abnormal seminiferous tubules, reduced sperm count and Leydig cell hyperplasia. Exposure to DBP could not induce the expression of VTG mRNA in either F₀ or F₁ male zebrafish, indicating that DBP has no estrogen-like effects on zebrafish. Conclusion Exposure to 1250 µg/L DBP can induce reproductive toxicity in F₀ and F₁zebrafish. The number of eggs and egg hatchability were reduced. Abnormal seminiferous tubules, reduced sperm count as well as Leydig cell hyperplasia were found.

Key words: Zebrafish, Dibutyl phthalate(DBP), Reproductive toxicity, F₀, F₁

中图分类号:

Q95-33

徐浩, 王玉柱, 温成丽, 成倩倩, 田芳, 夏敏杰, 李润生, 李卫华, 丁训诚. 邻苯二甲酸二丁酯对亲代和子代斑马鱼生殖毒性的比较研究[J]. 实验动物与比较医学, 2012, 32(5): 383-389.

XU Hao, WANG Yu-zhu, WEN Cheng-li, CHENG Qian-qian, TIAN Fang, XIA Min-jie, LI Run-sheng, LI Wei-hua, DING Xun-cheng. Reproductive Toxicity of Dibutyl Phthalate in Parent and Progeny Zebrafish[J]. Laboratory Animal and Comparative Medicine, 2012, 32(5): 383-389.

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邻苯二甲酸二丁酯对亲代和子代斑马鱼生殖毒性的比较研究

Reproductive Toxicity of Dibutyl Phthalate in Parent and Progeny Zebrafish

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