摘要
目的探究先天性低促性腺激素性性腺功能减退症(CHH)中成纤维细胞生长因子受体1(FGFR1)杂合缺失对生殖功能的作用。方法构建FGFR1杂合敲除小鼠模型(FGFR1^(+/-)),运用聚合酶链反应(PCR)与Sanger测序方法进行分析,验证目标外显子的敲除情况。采用免疫组化与Western blot,对蛋白表达进行检测。对小鼠的生殖能力、生殖功能(精子浓度、活力、前进性)展开测定,测量小鼠的BMI、睾丸质量、睾丸体积,以及各类性激素水平(包括T、E2、AMH、FSH、LH和INH-B),并进行统计学分析。结果研究构建了FGFR1杂合敲除的小鼠模型,PCR测序结果显示小鼠FGFR1外显子4、5被成功敲除,Western blot与免疫组化结果显示FGFR1蛋白表达减少。与野生型(WT)相比,FGFR1^(+/-)小鼠的精子活力更差(P<0.001),前进精子数量更少(P<0.01)。与WT小鼠相比,FGFR1^(+/-)小鼠的INH-B水平略有升高(P<0.05)。结论FGFR1杂合缺失影响小鼠精子活力,为深入探究先天性低促性腺激素性性腺功能减退症生殖功能障碍的发病机制提供了关键依据。
Objective To investigate the effect of heterozygous deletion of fibroblast growth factor receptor 1(FGFR1)on reproductive function in congenital hypogonadotropic hypogonadism(CHH).Methods A heterozygous FGFR1 knockout mouse model(FGFR1^(+/-))was established.Polymerase chain reaction(PCR)and Sanger sequencing were performed to confirm the deletion of the target exons.Immunohistochemistry and Western blot were used to detect FGFR1 protein expression.Reproductive capacity and function(including sperm concentration,motility,and progressive motility)were evaluated.Additionally,body mass index(BMI),testicular weight,testicular volume,and various sex hormone levels(including T,E2,AMH,FSH,LH,and INH-B)were measured and analyzed statistically.Results The FGFR1 heterozygous knockout mouse model was successfully constructed.PCR and sequencing results confirmed the deletion of exons 4 and 5 of the FGFR1 gene.Western blot and immunohistochemistry demonstrated reduced FGFR1 protein expression.Compared with wild-type(WT)mice,FGFR1^(+/-)mice exhibited significantly lower sperm motility(P<0.001)and fewer progressively motile sperm(P<0.01).Compared with WT mice,the INH-B level in FGFR1^(+/-)mice was slightly elevated(P<0.05).Conclusions Heterozygous deletion of FGFR1 impairs sperm motility in mice,providing critical evidence for further elucidating the pathogenesis of reproductive dysfunction in congenital hypogonadotropic hypogonadism.
作者
李晨阳
李婷
王曦
聂敏
伍学焱
茅江峰
韩钦
LI Chenyang;LI Ting;WANG Xi;NIE Min;WU Xueyan;MAO Jiangfeng*;HAN Qin(Department of Endocrinology,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100730;Center for Excellence in Tissue Engineering,Beijing Key Laboratory of Artificial Intelligence and Cell-based Medical Engineering for Interdisciplinary Innovation and Clinical Translation,Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100005,China)
出处
《基础医学与临床》
2026年第4期524-530,共7页
Basic and Clinical Medicine
基金
北京市自然科学基金(7212080)。
