小鼠睾丸组织中核受体4A1的表达变化及体外对睾丸间质细胞睾酮合成的影响  

Expression changes of nuclear receptor 4A1 in mouse testis and its effect on testosterone synthesis of Leydig cells in vitro

作　　者：谷亚龙 张新东[1] 金保方[1] 

机构地区：[1]南京中医药大学男科研究所,南京210046

出　　处：《山东医药》2015年第15期17-19,共3页Shandong Medical Journal

基　　金：国家自然科学基金青年基金资助项目(81302969);国家自然科学基金资助项目(81273760);国家自然科学基金资助项目(81473678);南京中医药大学青年自然科学基金项目(12XZR13)

摘　　要：目的观察核受体(NR)4A1在不同鼠龄小鼠睾丸组织中的表达变化及NR4A1对体外小鼠睾丸间质细胞睾酮合成的影响,并探讨其机制。方法 1周龄、4周龄、12周龄、12月龄雄性小鼠各6只,分别为幼年组、性发育前组、性成熟组及老年组。饲养3 d,适应环境后,各组小鼠全部脱颈处死,取双侧新鲜睾丸组织,采用real-time PCR法检测NR4A1 mRNA,Western blotting法检测NR4A1蛋白。体外培养小鼠睾丸间质细胞系MLTC-1,分别将NR4A1干涉腺病毒(Ad-si NR4A1)及空载腺病毒(Ad-CMV)转染MLTC-1。采用放射免疫法检测MLTC-1细胞培养液上清中的睾酮,real-time PCR法检测MLTC-1中的类固醇快速转运蛋白(St AR)、3β-羟类固醇脱氢酶(HSD3B)mRNA,Western blotting法检测St AR、HSD3B蛋白。结果幼年组、性发育前组、性成熟组、老年组小鼠睾丸组织中NR4A1 mRNA相对表达量以性发育前组最高,与余3组相比,P均<0.05;NR4A1蛋白相对表达量以性成熟期组最高,与余3组相比,P均<0.05。转染Ad-si NR4A1、Ad-CMV的MLTC-1中睾酮水平分别为(2.55±0.23)、(4.50±0.23)nmol/L,两者相比,P<0.05。转染Ad-si NR4A1的MLTC-1中St AR、HSD3B mRNA及其蛋白相对表达量与转染Ad-CMV的细胞相比,P均<0.05。结论不同鼠龄小鼠睾丸组织中NR4A1的表达有所差别;NR4A1可能通过调控St AR、HSD3B表达从而参与睾酮的合成。Objective To observe the expression changes of nuclear receptor （ NR） 4A1 in mouse testis and its effect on testosterone synthesis of Leydig cells in vitro, and to explore its mechanism.Methods The male mice of 1 week old, 4 weeks old, 12 weeks old and 12 months old were respectively divided into the infancy group, before sexual development group, sexual maturity group and the older group with 6 mice in each.After being fed for three days and adapting to the en-vironment, all the mice in each group were sacrificed.We took the fresh bilateral testicular tissue to detect NR4A1 mRNA by using real-time PCR, and NR4A1 protein by using Western blotting.We cultured the mouse Leydig cells MLTC-1 in vitro, and then used NR4A1 interference adenovirus （ Ad-siNR4A1 ） and no-load adenovirus （ Ad-CMV ） to transfect MLTC-1.The radioimmunoassay was used to measure the testosterone level in medium.real-time PCR and Western blotting were applied to determine the mRNA and protein expression of steroidogenic acute regulatory protein （ StAR ） and 3β-hydroxysteroid dehydrogenase （HSD3B）.Results The NR4A1 mRNA expression was the highest in the before sexual de-velopment group among the infancy group, before sexual development group, sexual maturity group and the older group;and significant difference was found as compared with the other three groups （all P〈0.05）.NR4A1 protein expression was the highest in the sexual maturity group as compared with that of the other three groups （ all P〈0.05） .The testoster-one levels in MLTC-1 transfected with Ad-siNR4A1 or Ad-CMV were respectively （2.55 ±0.23）, （4.50 ±0.23） nmol/L （all P〈0.05）.The StAR and HSD3B mRNA and protein expression levels in the MLTC-1 transfected with Ad-siNR4A1＆amp;nbsp;were relatively low as compared with those in the Ad-CMV-transfected MLTC-1 （all P〈0.05）.Conclusion NR4A1 ex-pression is different in testis tissues of mice with different age.NR4A1 is involved in the synthesis of testosterone possibly by regulating the

关 键 词：核受体4A1 睾丸 睾丸间质细胞 睾酮 3β-羟类固醇脱氢酶 类固醇快速转运蛋白 

分 类 号：R697[医药卫生—泌尿科学]