己烯雌酚促进实验动物卵泡发育同步化及其机制的初步研究

doi:10.3969/j.issn.1674-5817.2013.02.001

实验动物与比较医学 ›› 2013, Vol. 33 ›› Issue (2): 79-83.DOI: 10.3969/j.issn.1674-5817.2013.02.001

• 论著 • 下一篇

己烯雌酚促进实验动物卵泡发育同步化及其机制的初步研究

顾艳琼, 石燕, 王健, 孙兆贵

上海市计划生育科学研究所, 国家人口和计划生育委员会计划生育药具重点实验室, 上海市 200032

收稿日期:2012-06-07 出版日期:2013-04-25 发布日期:2013-04-25
作者简介:顾艳琼(1980-), 硕士。E-mail: yanqionggu@163.com
基金资助:
国家自然科学基金(81070490, 81170569)及上海市科学技术委员会项目(10140901900和10140901502)资助

Effects and Preliminary Mechanisms for Diethylstilbestrol on Follicular Development in Prepubertal Mice and Rats

GU Yan-qiong, SHI Yan, WANG Jian, SUN Zhao-gui

Key Laboratory of Contraceptive Drugs & Devices of NPFPC,Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China

Received:2012-06-07 Online:2013-04-25 Published:2013-04-25

摘要/Abstract

摘要： 目的观察人工合成的非甾体化合物己烯雌酚(DES)对性成熟前小鼠和大鼠卵巢中卵泡发育的影响,以及出现有卵泡同步化机理。方法取性成熟前雌性小鼠和大鼠,腹腔注射一定剂量的DES,然后取卵巢经连续切片和HE染色后,统计整个卵巢内三级卵泡数量。同时,取卵巢利用免疫组织化学和免疫印迹方法,分别检测泛素末端水解酶L1(UCH-L1),c-Jun结合蛋白1(Jab1)和细胞周期素依赖激酶抑制因子p27^Kip1在小鼠卵巢中的定位和含量。结果 UCH-L1定位在卵巢中各级卵泡的卵母细胞中,Jab1分布于次级和三级卵泡的卵母细胞核中,而p27^Kip1广泛分布于卵巢各类细胞中。免疫组化结果显示,仅就卵母细胞而言,由于DES的处理,UCH-L1和Jab1的表达信号增强,而p27^Kip1的信号变弱,与肿瘤细胞中的情况类似。然而,免疫印迹分析结果表明,以上3种蛋白在卵巢中相对β-actin的含量因DES处理而均呈现一定程度的上升变化趋势。青春前期的小鼠,在1~4周龄期间,卵巢中UCH-L1和p27^Kip1的蛋白量随着周龄增加的趋势,而Jab1的变化趋势不明显。结论腹腔注射DES或导致卵巢内卵泡发育同步化,其中涉及的机理可能是,UCH-L1及其关联蛋白Jab1导致p27^Kip1 在卵母细胞核中的含量下降,进而导致了多个卵泡的卵母细胞同时发育启动。

关键词: 己烯雌酚(DES), 青春期, UCH-L1, 卵泡同步发育

Abstract: Objective To observe the effects of the synthetic non-steroidal compound diethylstilbestrol on the ovarian folliclar development in the prepubertal mice and rats, and the involved mechanisms in the follicular synchronization resulted from the diethylstilbestrol treatment. Methods The prepubertal mice and rats were treated by intraperitoneal injection of a certain dose of diethylstilbestrol, and then the ovaries were collected to count the number of tertiary follicles in the ovary by serial sectioning and HE staining. On the other hand, the immunohistochemistry and immunoblotting methods were used to detect the distribution and quantity of Ubiquitin Carboxyl Terminal Hydrolases L1 (UCH-L1), c-Jun activation domain binding protein1 (Jab1), and cyclin dependent kinase inhibitive factor (p27^Kip1) in the ovaries, separately. Results Immunohistochemical results showed that the localization of UCH-L1 was only restricted into oocytes, Jab1 was detected in the oocyte nuclei of secondary follicles and tertiary follicles, and p27^Kip1 was distributed extensively in the ovary. Only considering the oocyte, in response to DES treatment, UCH-L1 and Jab1 staining signals were enhanced, and p27^Kip1 signal was become weak in the nucleus, similar to the situation in the tumor cells. However, the protein levels of all UCH-L1, Jab1 and p27^Kip1 in ovaries were showed increasing tendency in the DES-treated group to some degrees compared with naive group. In similarly rising tendency, the protein levels of UCH-L1 and p27^Kip1 were changed with the age in the ovaries of 1-4 week old mice though Jab1 was found no obvious change. Conclusion Intraperitoneal injection of diethylstilbestrol caused ovarian follicular growth synchronization, and involved mechanisms may be that UCH-L1 and its associated protein Jab1 down-regulated p27^Kip1 in the oocytes nucleus during the diethylstilbestrol-induced follicular growth synchronization.

Key words: Diethylstilbestrol, Puberty, UCH-L1, Follicle synchronization

中图分类号:

Q95-33

顾艳琼, 石燕, 王健, 孙兆贵. 己烯雌酚促进实验动物卵泡发育同步化及其机制的初步研究[J]. 实验动物与比较医学, 2013, 33(2): 79-83.

GU Yan-qiong, SHI Yan, WANG Jian, SUN Zhao-gui. Effects and Preliminary Mechanisms for Diethylstilbestrol on Follicular Development in Prepubertal Mice and Rats[J]. Laboratory Animal and Comparative Medicine, 2013, 33(2): 79-83.

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己烯雌酚促进实验动物卵泡发育同步化及其机制的初步研究

Effects and Preliminary Mechanisms for Diethylstilbestrol on Follicular Development in Prepubertal Mice and Rats

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