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.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
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)
