hCG调节原代培养小鼠睾丸Leydig细胞Atrn mRNA和Atrn蛋白的表达及与睾酮分泌的相关性  

HCG- stimulated expression of Atrn mRNA and Atrn protein in primary cultured mouse Leydig cells and their relationships to testosterone production

作　　者：明钰[1] 熊承良[2] 

机构地区：[1]武汉轻工大学医学技术与护理学院,湖北武汉430023 [2]华中科技大学同济医学院计划生育研究所,湖北武汉430022

出　　处：《遵义医学院学报》2014年第1期86-88,93,共4页Journal of Zunyi Medical University

基　　金：国家自然科学基金资助项目(NO:30570679)

摘　　要：目的探讨hCG刺激的Leydig细胞睾酮分泌与Atrn mRNA和Atrn蛋白表达的关系。方法将原代培养的Leydig细胞分为5组,分别在培养液中加入0、0.1、1、10、100 ng/mL hCG,培养24 h后吸取培养液用于测定睾酮分泌量。同时提取细胞mRNA和总蛋白,用实时定量RT-PCR和Western blot方法分别检测Leydig细胞中Atrn mRNA和Atrn蛋白表达量。结果刺激24 h后,与0 ng/mLhCG剂量组相比较,0.1,1,10,100 ng/mL hCG剂量组睾酮生成量都明显增加,统计学差异较大(P<0.01)。Leydig细胞Atrn mRNA表达量增加(0.1 ng/mL hCG组P<0.05,其余剂量组P<0.01)。睾酮生成量与Atrn蛋白表达量(r=0.987,P<0.01)和Atrn mRNA表达量(r=0.982,P<0.01)呈正相关。结论原代培养的Leydig细胞在hCG刺激下,睾酮生成量、Atrn mRNA和Atrn蛋白表达量都呈现hCG剂量依赖性的增加。Objective To investigate the hCG -stimulated expression of Atrn mRNA and Atrn protein in primary cultured mouse Leydig cells and their relationships to testosterone production. Methods The primary cultured mouse Leydig cells were divided into five groups. The cells were stimulated by hCG at a concentration of 0, 0.1, 1, 10, 100 ng/ml separately. Culture media were collected and used for the assay of testosterone after 24h incu- bation. The treated Leydig cells were used for the measurement of mRNA and protein for Atrn by real - time RT -PCR and Western blot. Results Treatment of the Leydig cell cultures with different doses of hCG （0 - l0 ng/ ml） produced a dose - response induction curve of testosterone secretion, Atrn mRNA and Atrn protein expres- sion. Multiple correlation analysis showed that there was a positive correlation between the protein expression lev- els of Atrn and level of testosterone （ r = 0. 987, P 〈 0.01 ）, and a positive correlation between the gene expres- sion of Atrn and testosterone levels （ r = 0. 982, P 〈 0.01 ）. Conclusion The expression of Atrn mRNA and Atrn protein as well as the testosterone production were significantly stimulated by hCG at different doses of hCG （0 10 ng/ml） in Leydig cells.

关 键 词：小鼠 HCG ATTRACTIN LEYDIG细胞 睾酮 

分 类 号：Q492.4[生物学—生理学] Q813.1