己烯雌酚促验进实动物卵泡发育同步化及其机制的初步研究

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LI（UCH-L1），c-Jun结合蛋白l（Jabl）和细胞周期素依赖激酶抑制因子p27KipI在小鼠卵巢中的定位和含量。结果UCH-LI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p27KipI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p27KipZ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b-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p27KiPI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p27KiPI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。

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

Objective To observe the effects of the synthetic non-steroidal compound diethylstil- bestrol on the ovarian folliclar development in the prepubertal mice and rats, and the involved mecha- nisms 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 proteinl （Jabl）, and cyclin dependent kinase inhibitive factor （p27KiPI） in the ovaries, separately. Results Immunohistochemical results showed that the localization of UCH-L1 was only restricted into oocytes, Jabl was detected in the oocyte nuclei of secondary follicles and tertiary follicles, and p27rap1 was distributed extensively in the ovary. Only considering the oocyte, in response to DES treatment, UCH-L1 and Jabl staining signals were enhanced, and p27KiPI signal was become weak in the nucleus, similar to the situation in the tumor cells. However, the protein levels of all UCH-L1, Jabl and p27~apl 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 p27rapt were changed with the age in the ovaries of 1-4 week old mice though Jabl 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 Jabl down-regulated p27KiPI in the oocytes nucleus during the diethylstilbestrol-induced follicular growth synchronization.