Geng Xu-Jing, Zhao Dong-Mei, Mao Gen-Hong, Tan Li
Reproductive Medical Center, The Second Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Reproduction. 2017 Sep;154(3):229-236. doi: 10.1530/REP-17-0234. Epub 2017 Jun 13.
Leydig cells are essential for male reproductive development throughout life. Production of androgens as well as intermediate steroids is tightly regulated. Although microRNAs (miRNAs) are suggested to play important roles in spermatogenesis, little is currently known regarding the regulation of steroidogenesis by miRNAs in Leydig cells. Here, we found that miR-150 was predominantly expressed in Leydig cells within mouse testis. Therefore, we determined steroidogenesis of the Leydig cells in which miR-150 was knocked down or overexpressed using miR-150 antagomir and agomir, respectively. Compared with negative control group, a significant increase of STAR expression was observed in miR-150 antagomir-treated Leydig cells. Conversely, STAR expression was significantly reduced in miR-150 agomir-transfected Leydig cells. Production of sex-steroid precursors and testosterone of Leydig cells was also negatively controlled by miR-150. We further identified as a target of miR-150 using luciferase reporter assay. Finally, we confirmed that miR-150 was necessary for steroidogenesis and spermatogenesis via intratesticular injection of miR-150 antagomir or agomir. Taken together, our studies suggest that miR-150 negatively regulates the expression of STAR and steroidogenesis of Leydig cells in mice.
睾丸间质细胞对于男性一生的生殖发育至关重要。雄激素以及中间类固醇的产生受到严格调控。尽管微小RNA(miRNA)被认为在精子发生中发挥重要作用,但目前关于miRNA对睾丸间质细胞类固醇生成的调控知之甚少。在此,我们发现miR-150在小鼠睾丸的间质细胞中大量表达。因此,我们分别使用miR-150拮抗剂和激动剂,测定了miR-150敲低或过表达的间质细胞的类固醇生成情况。与阴性对照组相比,在经miR-150拮抗剂处理的间质细胞中观察到类固醇生成急性调节蛋白(STAR)表达显著增加。相反,在转染了miR-150激动剂的间质细胞中,STAR表达显著降低。间质细胞的性类固醇前体和睾酮的产生也受到miR-150的负调控。我们通过荧光素酶报告基因检测进一步确定[具体内容缺失]为miR-150的一个靶点。最后,我们通过睾丸内注射miR-150拮抗剂或激动剂证实,miR-150对于类固醇生成和精子发生是必需的。综上所述,我们的研究表明,miR-150负向调节小鼠间质细胞中STAR的表达和类固醇生成。