低剂量棉酚与甾体激素联合应用对大鼠睾丸Cdc25A、cyclinB1及雄激素受体蛋白表达的影响  

Effect of low-dose gossypol acetic acid combined with steroid hormones on the expressions of Cdc25A,cyclin B1 and androgen receptor in adult rat testis

作　　者：张凤枝[1,2] 刘昉[1,2] 许增禄[1,2] 钱晓菁[1,2] 邵金辉[1,2] 黄卉[1,2] 

机构地区：[1]中国医学科学院基础医学研究所 [2]北京协和医学院基础医学院人体解剖学与组织胚胎学系,北京100005

出　　处：《解剖学报》2010年第3期440-445,共6页Acta Anatomica Sinica

摘　　要：目的探讨低剂量棉酚与甾体激素联合应用(棉甾联合用药)对大鼠睾丸Cdc25A、cyclinB1及雄激素受体(AR)蛋白表达的影响。方法采用大鼠喂服棉酚12.5mg/(kg.d)+激素[去氧孕烯125μg/(kg.d)+炔雌醇25μg/(kg.d)+十一酸睾丸酮100mg/(kg.d)]联合用药的方式,与单独喂服相同剂量的棉酚或激素的大鼠及喂服甲基纤维素溶剂的大鼠相对照,通过免疫组织化学染色和免疫印迹法,观察Cdc25A、CyclinB1及AR蛋白表达的变化。结果各时间点(6周、8周和10周)激素组和棉甾联合用药组Cdc25A在顶体和间质中的表达较对照组减弱,均有显著性差异(P<0.05)。用药6周和8周时,棉酚组、激素组和棉甾联合用药组Cdc25A在精母细胞中表达较对照组增强(P<0.05);用药10周后,只有激素组显著下降(P<0.05)。各时间点激素组和棉甾联合用药组中CyclinB1在间质细胞中的表达量明显降低(P<0.05),但在圆形精子细胞中的表达量无明显改变。各时间点激素组和棉甾联合用药组间质细胞的AR阳性出现在细胞核,而对照组定位于细胞质。Western blotting结果显示,各用药组大鼠睾丸组织中Cdc25A的表达量与免疫组织化学染色结果的变化趋势基本一致。CyclinB1的含量在各组中无明显差异。结论棉甾联合用药影响Cdc25A、cyclin B1和AR蛋白在生精细胞和Leydig细胞中的表达,进而影响减数分裂和圆形精子的变态过程;甾体激素的重要作用是抑制间质细胞的功能发挥。Objective To investigate the expressions of Cdc25A, cyclin B1 and androgen receptor （AR） by administering with combination regimen of low-dose gossypol acetic acid （GA） with steroid hormones [ desogestrel/ ethinylestradiol/testosterone undeeanote （DSG/E/TU）] in adult rat testis, Methods Adult male rats were randomly divided into four groups. Group GH ： rats were fed orally with GA [ 12. 5 mg/（ kg · day） ] and DSG [ 125μg/（ kg · day） ]/E [ 25 μg/（ kg · day） ]/TU [ 100mg/（ kg · day） ] ; Group G ： rats were administered with a single dose of GA [ 12.5 rag/（ kg · day） ] ; Group H ： DSG [ 125 p,g/（ kg · day ） ]/E [ 25 ~g/（ kg · day） ]/TU [ 100mg/（ kg · day ） ] were administered to rats ; Group C： rats only received vehicle （1% methyl cellulose）. The testes were excised at the 6th, 8th and lOth week, respectively. The expressions of Cde25A, cyclin B1 and AR are detected by immunohistochemical staining and Western blotting. Results Compared with the control group, the intensities of Cdc25A immunohistochemieal positive staining were decreased in Leydig cells and acrosomes at the 6th,8th and 10th week（ P 〈 0. 05 ） , and increased in spermatocytes in G, H and GH group at the 6th and 8th week （P 〈 0. 05）. The intensities of cyclin B1 immunohistochemical positive staining were decreased in Leydig cells in H and GH group at the 6th,8th and 10th week（P 〈 0. 05 ）. AR positive staining appeared in the nuclei of Leydig cells in H and GH group instead of the cytoplasm. Western blotting showed that the expression of Cdc25A was similar with immunohistoehemistry results generally. The expression of cyclin B1 in rat testis in each group was steady. Conclusion The expressions of Cdc25A, cyclin B1 and AR were altered by administering with combination regimen of low-dose gossypol acetic acid with steroid hormones in the testis of adult rat, by which the meiosis and the spermiogenesis of the spermatogenesis process might be disturbed. Steroid hormones migh

关 键 词：棉酚 甾体激素 CDC25A CYCLINB1 雄激素受体 生精细胞 免疫组织化学 免疫印迹法 大鼠 

分 类 号：R979.2[医药卫生—药品]