UT-B基因敲除小鼠睾丸间质细胞睾酮分泌水平变化及其对性成熟的影响  

Change of testosterone secretion level of Leydig cells of UT-B knockout mice and its influence in sexual maturity

作　　者：赵丽晶[1] 路明 韩雪[1] 张睿[1] 李若文 赵雪俭[1] 杨宝学[1,4] 郭丽荣[1] 

机构地区：[1]吉林大学白求恩医学院病理生理学系,吉林长春130021 [2]黑龙江省五常市人民医院普外科,黑龙江五常150200 [3]吉林亚泰生物药业股份有限公司 [4]北京大学医学部药理学系,北京100191

出　　处：《吉林大学学报（医学版）》2013年第3期448-452,F0002,共6页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金青年基金资助课题(81000271)

摘　　要：目的:利用尿素通道蛋白B(UT-B)基因敲除小鼠模型探讨UT-B基因对雄性小鼠睾丸间质Leydig细胞分泌睾酮能力的影响。方法:饲养、繁殖并鉴定UT-B基因敲除(UT-B-/-)小鼠;选取4周龄遗传背景相同的野生型(UT-B+/+)雄性小鼠和UT-B-/-雄性小鼠各5只,采用放射免疫法检测2种小鼠血清睾酮和黄体生成素(LH)水平;分离培养并鉴定Leydig细胞,测定培养液上清中睾酮的水平。结果:成功获得UT-B-/-雄性小鼠和UT-B+/+雄性小鼠;成功分离培养原代Leydig细胞并经3β-HSD染色方法鉴定纯度达80%以上;4周龄UTB-/-雄性小鼠血清睾酮水平[(7.29±1.27)×10-2μg.L-1]明显高于UT-B+/+雄性小鼠[(5.53±1.58)×10-2μg.L-1](P<0.05),2种小鼠血清LH水平比较差异无统计学意义(P>0.05);UTB-/-雄性小鼠Leydig细胞培养上清中睾酮水平[(17.300±1.179)nmol.L-1]明显高于UT-B+/+雄性小鼠[(12.300±0.916)nmol.L-1](P<0.01)。结论:4周龄UT-B-/-雄性小鼠Leydig细胞分泌睾酮的能力明显高于UT-B+/+小鼠,这可能是UT-B-/-雄性小鼠比UT-B+/+小鼠生殖育龄提前的原因之一。Objective To investigate the effect of urea transporter B（UT-B） gene on testosterone secreted by Leydig cells of male mice by using UT-B gene knockout mouse models.Methods The UT-B gene knockout（UT-B-/-） mice were fed,bred and identified.5 UT-B wild type（UT-B＋/＋） male mice and 5 UT-B-/-male mice from the same genetic background aged 4 weeks were selected.The levels of the serum testosterone and luteotropic hormone（LH） of two kinds of mice were detected by radioimmunoassay;the Leydig cells of two kinds of mice were isolated and cultured and then identified;the level of testosterone in culture supernatants was detected.Results The UT-B＋/＋ male mice and UT-B-/-male mice were obtained successfully;the primary Leydig cells were isolated and cultured and the cell purity was up to 80% identified by 3β-HSD staining method.The serum testosterone level of UT-B-/-male mice aged 4 weeks [（7.29±1.27）×10-2 μg·L-1]was higher than that of UT-B＋/＋ male mice[（5.53±1.58）×10-2 μg·L-1]（P0.05）,but there was no significant difference of serum LH levels between two kinds of mice（P0.05）.The testosterone level in the Leydig cells supernatants of UT-B-/-male mice[（17.300±1.179） nmol·L-1] was higher than that of UT-B＋/＋ male mice[（12.300±0.916） nmol·L-1]（P0.01）.Conclusion The ability of Leydig cells secreting testosterone of UT-B-/-male mice aged 4 week is stronger than that of UT-B＋/＋ mice,which might be one of the reasons that the reproductive age of UT-B-/-male mice is earlier than that of UT-B＋/＋ male mice.

关 键 词：尿素通道蛋白B 睾丸间质细胞 睾酮 基因敲除 

分 类 号：R318.15[医药卫生—生物医学工程]