度他雄胺对大鼠附睾Claudin1和β-catenin表达的影响  

Inhibitory effect of dutasteride on the expressions of epididymal Claudin1 and β-catenin in male rats

作　　者：谢淑武[1] 曲立娟[2] 周娴颖[1] 周洁芸[1] 李国停 毕继宏 郭湘洁[1] 李钊[1] 曹霖[1] 朱焰[1] 

机构地区：[1]上海市计划生育科学研究所生殖药理研究组国家卫生与计划生育委员会计划生育药具重点实验室,上海200032 [2]上海市第八人民医院检验科,上海200235 [3]广东医学院药学院,广东湛江524000

出　　处：《中华男科学杂志》2015年第1期17-22,共6页National Journal of Andrology

基　　金：国家自然科学基金(81100463)~~

摘　　要：目的：通过度他雄胺对大鼠附睾上皮连接关键蛋白Claudin1和β-catenin表达影响的研究,探讨度他雄胺抑制生育的可能分子机制,为男性不育治疗和男性避孕提供附睾靶点。方法：16只SD雄性大鼠（3月龄,250~300 g）,设度他雄胺40 mg/kg实验组和阴性对照组,每组8只,灌胃给药,1次/d,连续2周。给药结束后计算机辅助精子分析雄鼠精子活力及形态,计算交配雌鼠受孕率,ELISA法测定大鼠血清睾酮（T）和双氢睾酮（DHT）浓度,透射电镜分析附睾细胞紧密连接变化,RT-PCR分析实验组和对照组附睾上皮连接蛋白Claudin1和β-catenin基因表达变化;免疫组化和免疫印迹分析其蛋白表达变化。结果：度他雄胺可以显著抑制大鼠血清DHT浓度、精子活力和生育力,超微结构显示实验组附睾上皮细胞紧密连接出现空隙,附睾腔液体积增多。RT-PCR、免疫组化和免疫印迹显示度他雄胺可以显著下调Claudin1和β-catenin的表达。结论：度他雄胺可以显著抑制雄性大鼠生育,这可能与其降低大鼠血清DHT水平,进而抑制附睾上皮连接关键基因Claudin1和β-catenin表达和破坏附睾上皮紧密连接有关。Objective： To explore the molecular mechanism of dutasteride inhibiting fertility by studying its effects on the expressions of the epididymal epithelial junction proteins Claudin1 and β-catenin in rats. Methods： Sixteen 3-month-old SD male rats were equally divided into an experimental and a negative control group to be treated intragastrically with dutasteride at 40 mg / kg per day and the same dose of solvent,respectively,for 14 consecutive days. Then,the sperm motility and morphology of the rats were detected by computer-assisted sperm analysis,the serum levels of testosterone（ T） and dihydrotestosterone（ DHT） measured by ELISA,changes in the tight junction of epididymal cells observed under the transmission electron microscope,the protein and gene expressions of Claudin1 and β-catenin determined by RT-PCR and immunohistochemistry,and the conception rate of the mated female rats calculated. Results： Dutasteride significantly suppressed the serum DHT level,sperm motility,and fertility of the rats（ P 〈 0. 05）. Interspaces between epididymal epithelial cell tight junctions were observed,the volume of epididymal fluid obviously increased,and the expressions of Claudin1 and β-catenin gene and protein remarkably downregulated in the experimental rats（ P 〈 0. 05）. Conclusion：Dutasteride can significantly inhibit the fertility of male rats by reducing the serum DHT level,suppressing Claudin1 and β-catenin expressions,and damaging epididymal epithelial cell junctions.

关 键 词：度他雄胺 双氢睾酮 附睾上皮连接 Claudin1 Β-CATENIN 

分 类 号：Q492.4[生物学—生理学] R965.2[医药卫生—药理学]