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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 effect of combination of low dose gossypol and steroid hormone (MCS) on the expression of Cdc25A, cyclinB1 and androgen receptor (AR) in rat testes. Methods The rats were fed with gossypol 12.5 mg / (kg · d) + hormones [desogestrel 125 μg / (kg · d) + ethinyl estradiol 25 μg / (kg · d) + testosterone undecanoate 100 mg / (kg .d)] combined with the way, with the same dose of gossypol or hormone alone fed rats and rats fed methylcellulose solvent contrast, by immunohistochemical staining and Western blotting to observe the Cdc25A, Changes of CyclinB1 and AR protein expression. Results The expression of Cdc25A in the acrosome and interstitium at different time points (6, 8 and 10 weeks) was significantly lower than that in the control group (P <0.05). At 6 weeks and 8 weeks after treatment, the expression of Cdc25A in gossypol, hormones and gonaditis groups was higher than that in control group (P <0.05). After 10 weeks of treatment, only hormone group was significantly decreased (P < 0.05). At each time point, the expression of CyclinB1 in the interstitial cells was significantly decreased (P <0.05), but there was no significant change in the round sperm cells. At each time point, the AR positive of interstitial cells in the hormone group and the gonadal drug combination group appeared in the nucleus, while the control group was located in the cytoplasm. Western blotting results showed that the expression levels of Cdc25A in testis tissue of rats in each treatment group were consistent with those of immunohistochemical staining. CyclinB1 content in each group no significant difference. Conclusion The combination of gonadal injection and Cdc25A, cyclin B1 and AR protein expression in spermatogenic cells and Leydig cells, thereby affecting meiotic and round sperm metamorphosis; steroid hormone’s important role is to inhibit the function of interstitial cells Play