摘要
目的研究一氧化氮(NO)供体(硝普钠,SNP)对大鼠睾丸间质细胞分泌睾酮的影响及其机制。方法大鼠睾丸间质细胞原代培养12 h后,用不同浓度的SNP(0,10-8,10-6,10-4mol/L)分别处理4、8、12 h,用化学发光法检测细胞上清睾酮(T)水平的变化;western blot法分析间质细胞内3β-HSD(3β-羟基类固醇脱氢酶)和annexin 5(膜联蛋白5)的变化。结果10-4mol/L的SNP处理间质细胞8 h后,睾酮水平较对照组上升23%(P<0.05);10-6mol/L的SNP处理12 h后,睾酮水平较对照组上升28%(P<0.05),10-4mol/L的SNP处理12 h,较对照组升高37%(P<0.05)。western blot结果显示,SNP的刺激对3β-HSD表达无显著影响。不同浓度SNP刺激间质细胞4 h和8 h后,annexin 5表达量与对照组比较无显著差异(P>0.05);10-6mol/L的SNP刺激12 h后,annexin 5表达量达到最高水平,较对照组升高55%(P<0.05);10-4mol/L的SNP处理12 h,annexin 5表达量较对照组升高48%(P<0.05)。结论SNP在低浓度范围内促进大鼠睾丸间质细胞分泌睾酮,且睾酮的分泌对SNP具有时间和剂量的依赖性;SNP刺激睾酮分泌的过程并非通过3β-HSD介导,但可能与annexin 5表达有关。
Objective To study the effect of NO donor （sodium nitroprusside, SNP） on testosterone secretion in rat Leydig cells. Methods The primary rat Leydig cells were cultured in vitro for 12 h, and then treated with different concentrations of SNP for various time courses （4 h, 8 h, and 12 h）. The level of testosterone was detected by chemiluminescence, and the concentrations of 3β- hydroxysteroid dehydrogenase enzyme （3β-HSD） and annexin 5 were analyzed by Western blotting. Results After treatment with 10^-4 mol/L SNP for 8 h, with 10^-6 mol/L SNP for 12 h, or with 10^-4 mol/L SNP for 12 h, the level of testosterone was increased by 23%, 28%, and 37% respectively, compared with that of the control （P 〈 0.05 ）. Western blot analysis showed that, when Leydig cells were stimulated by SNP, 3β-HSD expression had no significant difference （P 〉 0.05）, compared with that in the control. After Leydig cells were treated with different concentrations of SNP for 4 h or 8 h, the annexin 5 level also had no difference compared with that in the control （P 〉 0.05）. However, after Leydig cells were stimulated by 10^ -6 mol/L SNP for 12 h, the annexin 5 was increased by 55% compared with that of the control （P 〈 0.05）, and the expression of annexin 5 also increased by 48% in the cells treated with 10^-4 mol/L SNP （P 〈 0.05 ）. Conclusions The results suggest the low concentration of SNP induces testosterone secretion in rat Leydig cells in a dose-and time-dependent manner. 3β-HSD is not involved in the process of testosterone secretion induced by SNP,but the process may be related to the expression of annexin 5.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2010年第2期139-142,共4页
Chinese Journal of Clinical Laboratory Science
基金
国家自然科学基金项目(编号:30770801)
