hCG对成年小鼠睾丸Leydig细胞中睾酮生成酶的调节作用

THE REGULATORY EFFECT OF hCG ON STEROIDOGENIC ENZYMES IN ADULT MOUSE LEYDIG CELLS

目的 从分子水平探讨 h CG对成年小鼠睾丸 L eydig细胞中胆固醇侧链裂解酶 (P45 0 scc)、17α-羟化酶 (P45 0 c17)和 3β-羟甾脱氢酶 (3βHSD)的调节作用。 方法 体外培养成年小鼠睾丸 L eydig细胞 ,分别测定培养1、8h h CG刺激组及对照组细胞培养上清中睾酮含量 ,同时运用半定量 RT- PCR及核酸内切酶酶切方法测定两组细胞中 P45 0 scc、P45 0 c17及 I、VI两型总的 3βHSD的 m RNA水平 ,并测定了 I、VI两型 3βHSD m RNA的比值。 结果 1.培养 1、8h刺激组睾酮含量均明显高于对照组 (1h P<0 .0 1,8h P<0 .0 0 5 ) ;2 .培养 8h,刺激组 P45 0 scc和P45 0 c17m RNA水平明显高于对照组 (P<0 .0 5 ) ;而培养 1、8h刺激组 I、VI两型总的 3βHSD m RNA水平和对照组相比均无明显差异 (P>0 .0 5 ) ,但 VI型 3βHSD m RNA和 I型 3βHSD m RNA的比值逐渐增高。 结论 h CG能在转录水平刺激 P45 0 scc、P45 0 c17及 VI型 3βHSD的表达 ,促进睾酮生成。

Objective To study the regulation role on molecular level of hCG on P450scc,P450c17 and 3βHSD in adult mouse Leydig cells. Methods Leydig cells of adult mice were cultured in vitro for 1 and 8 hours with or without hCG,then P450scc,P450c17 and I,VI total 3βHSD mRNA levels were measured respectively by the semi quantitative RT PCR as well as Scal restriction endonuclease incised methods.Meanwhile,the testosterone contents were measured in two groups. Results 1.In stimulated group Leydig cells cultured for 1 and 8 hours with hCG,the testosterone contents were higher significantly than those in control groups(1 hour P <0 01,8 hour P <0 005);2.After cultured for 8 hours,the P450scc and P450c17 mRNA levels in hCG stimulated cells were higher than those in control cells( P <0 05);the total level of 3βHSD mRNA had little difference with control group cells( P >0.05),but the ratio of type VI/I 3βHSD mRNA gradually increased. Conclusion hCG can stimulate the transcription of P450scc,P450c17 and type VI 3βHSD in adult mouse Leydig cells.