Dou Ke, Li Hong, Lu Yi-ping, Wei Qiang, Yang Yu-ru, Dong Qiang
Department of Urology, West China Hospital, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2009 Mar;40(2):284-7, 301.
To prepare the cDNA probe of the steroidogenic acute regulatory (StAR) gene, and to investigate the StAR mRNA expression in stressed C57BL/6 mice leydig cells.
cDNA fragment encoding mouse StAR was amplified by RT-PCR from the total RNA prepared from the testis, and then the RT-PCR product was cloned into pCR2. 1-TOPO vector. After StAR gene cDNA was sequenced, the Dig-labeled cRNA probes for mouse StAR gene were prepared by in vitro transcription from cDNA fragment. With the specific cRNA probes, in situ hybridization analysis was conducted in stressed mice testis and controls.
The results demonstrated that StAR mRNA levels were significantly lower in stressed leydig cells than that in controls (P < 0.05).
The decreased StAR mRNA levels induced by stress may result in a reduced production of StAR protein, which compromised the transportation of cholesterol, the substrate for androgen biosynthesis, into mitochondria, resulting in a poor T production in leydig cells and finally a declined T levels.
制备类固醇生成急性调节蛋白（StAR）基因的cDNA探针，并研究应激状态下C57BL/6小鼠睾丸间质细胞中StAR mRNA的表达。
从睾丸制备的总RNA中通过RT-PCR扩增编码小鼠StAR的cDNA片段，然后将RT-PCR产物克隆到pCR2.1-TOPO载体中。对StAR基因cDNA进行测序后，通过从cDNA片段进行体外转录制备用于小鼠StAR基因的地高辛标记的cRNA探针。使用特异性cRNA探针，对应激小鼠睾丸和对照进行原位杂交分析。
结果表明，应激状态下睾丸间质细胞中StAR mRNA水平显著低于对照组（P<0.05）。
应激诱导的StAR mRNA水平降低可能导致StAR蛋白产生减少，这损害了雄激素生物合成底物胆固醇向线粒体的转运，导致睾丸间质细胞中睾酮生成减少，最终导致睾酮水平下降。
