外源雄激素促进绵羊附睾GPx5基因的表达

Promoting effect of exogenous androgen on the expression of GPx5 gene in sheep epididymis

为研究外源雄激素丙酸睾酮对绵羊附睾特异表达基因谷胱甘肽过氧化酶5(GPx5)的影响,应用酶联免疫吸附(ELISA)法检测血清和附睾头部内睾酮(T)、二氢睾酮(DHT)的含量,及附睾头部雄激素受体(AR)蛋白表达量;应用实时荧光定量PCR(RT-PCR)技术检测了附睾头部GPx5-mRNA和AR-mRNA相对表达量;并结合免疫印迹(WB)法检测了附睾头部GPx5蛋白相对表达量。结果表明:丙酸睾酮处理(TPT)组与对照组相比绵羊血清DHT含量,附睾头部T含量、DHT含量均显著提高,分别是对照组的0.5、6.0、0.8倍;GPx5-mRNA和ARmRNA相对表达量显著提高,分别是对照组的0.75和0.77倍(P<0.05);AR及GPx5蛋白表达量显著提高,分别是对照组的0.63和0.58(P<0.05)。因此,外源雄激素丙酸睾酮可大量提高绵羊附睾头部GPx5基因的表达,对绵羊附睾头部GPx5基因的表达具有促进作用。

To explore the effect of exogenous androgen testosterone propionate on the expression of epididymis caputspecific glutathione peroxidase 5gene（GPx5）in sheep,Enzyme-linked immunosorbent assay was adopted to detect the content of dihydrotestosterone（DHT）and testosterone（T）in serum and epididymal caput,and the expression level of androgen receptor（AR）protein in epididymal caput.Moreover,the relative expressions of GPx5-mRNA and AR-mRNA were measured by Real-time fluorescent quantitative PCR（RT-PCR）;and GPx5 protein expression were analyzed by Western blotting.The results showed that：DHT in serum and T,DHT in the caput epididymidis were significantly increased to 0.5,6.0and 0.8folds（P〈0.05）,respectively;GPx5-mRNA and AR-mRNA expression in epididymal caput were significantly upregulated to 0.75 and 0.77folds（P〈0.05）;the expression levels of GPx5 and AR proteins were also increased to 0.63 and 0.58folds（P〈0.05）in testosterone propionate treatment group compared with the control group.In conclusion,exogenous androgen testosterone propionate could promote GPx5 gene expression in sheep epididymal caput.