大鼠睾丸间质细胞中促性腺激素释放激素激动剂调控睾酮合成的机制
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国家自然科学基金项目(30770801)


Mechanism of GnRH agonist-induced testosterone synthesis in rat Leydig cells
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    目的:研究促性腺激素释放激素(GnRH)激动剂(GnRHa)对大鼠睾丸间质细胞睾酮合成的调控机制-方法:原代培养大鼠睾丸间质细胞,GnRHa (10-7 mol/L)分别处理间质细胞6-12-24-36-48 h后,Western blot方法分析3β-羟基类固醇脱氢酶(3β-HSD)蛋白的变化;MEK1/2抑制剂PD98059 (50 μmol/L) 和GnRHa共作用于间质细胞后,分析3β-HSD蛋白表达和睾酮水平的变化-结果:GnRHa处理间质细胞不同时间后,结果显示3β-HSD蛋白的表达对GnRHa的刺激具有时间依赖性-GnRHa处理12 h后,3β-HSD蛋白表达与对照组相比显著上升45% (P < 0.05),24 h达到最高水平,升高1.0倍(P < 0.05);48 h后3β-HSD恢复至正常水平-加入PD98059后,显著抑制了GnRHa对3β-HSD的刺激作用,3β-HSD水平下降了61% (与GnRHa组相比,P < 0.05),睾酮水平也随之显著下降45% (与GnRHa组相比,P < 0.05)-结论:GnRHa可能通过细胞外信号调节激酶(ERK1/2)途径调控大鼠睾丸间质细胞3β-HSD的表达,进而促进睾酮的合成-

    Abstract:

    Objective:To study regulation mechanism of GnRH agonist(GnRHa) on testosterone synthesis in rat Leydig cells. Methods:The primary rat Leydig cells were cultured 24 h in vitro, and then treated with GnRHa (10-7 mol/L) at varied time courses (6,12,24,36,48 h);the expression of 3β-HSD protein was analyzed by Western blot. After stimulation with GnRHa and PD98059 (50 μmol/L),the specific inhibitor of ERK1/2,in Leydig cells,the expression of 3β-HSD and the level of testosterone were also investigated. Results:Treatment with GnRHa resulted in 3β-HSD expression in a time-dependent manner. 3β-HSD expression was significantly increased 45% over control after 12 h of treatment with GnRHa (P < 0.05),maximally elevated 1.0-fold over control at 24 h (P < 0.05),and then returned to basic level after 48 h. We also found the activation of 3β-HSD induced by GnRHa was markedly reversed in the presence of PD98059 (61% compared with GnRHa group,P < 0.05),following the significant decrease of testosterone (45% compared with GnRHa group,P < 0.05). Conclusion:The results suggest that GnRHa regulates the expression of 3β-HSD through ERK1/2,and further induces testosterone synthesis.

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柳海燕,陶晓倩,时姗姗,张艳梅,卢坤刚,韩雪峰,崔英霞,戈一峰,黄宇烽,李晓军,姚〓兵.大鼠睾丸间质细胞中促性腺激素释放激素激动剂调控睾酮合成的机制[J].南京医科大学学报(自然科学版),2009,29(10):1337-1341

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  • 收稿日期:2009-04-07
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