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Expression of GDNF and AR in Testicular Peritubular Cells of Surgery-induced Cryptorchidism Mice
- Journal of Sun Yat-sen University(Medical Sciences) Vol. 45, Issue 1, Pages: 85-92(2024)
- 作者机构：
1.安徽医科大学附属省儿童医院泌尿外科，安徽 合肥 230051
2.安徽医科大学第五临床医学院泌尿外科，安徽 合肥 230051
- 作者简介：
CHAO Min； E-mail： cm0654@sina.com
- 基金信息：
DOI：10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).20240004.020
CLC： R726.9Published：20 January 2024，
Received：12 September 2023，
Accepted：03 December 2023
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吴飞,潮敏,张殷等.神经胶质细胞系衍生神经营养因子和雄激素受体在手术诱导隐睾小鼠睾丸管周细胞中的表达[J].中山大学学报（医学科学版）,2024,45(01):85-92.
WU Fei,CHAO Min,ZHANG Yin,et al.Expression of GDNF and AR in Testicular Peritubular Cells of Surgery-induced Cryptorchidism Mice[J].Journal of Sun Yat-sen University(Medical Sciences),2024,45(01):85-92.
吴飞,潮敏,张殷等.神经胶质细胞系衍生神经营养因子和雄激素受体在手术诱导隐睾小鼠睾丸管周细胞中的表达[J].中山大学学报（医学科学版）,2024,45(01):85-92. DOI： 10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).20240004.020.
WU Fei,CHAO Min,ZHANG Yin,et al.Expression of GDNF and AR in Testicular Peritubular Cells of Surgery-induced Cryptorchidism Mice[J].Journal of Sun Yat-sen University(Medical Sciences),2024,45(01):85-92. DOI： 10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).20240004.020.
摘要
目的
2
探讨神经胶质细胞系衍生神经营养因子（GDNF）和雄激素受体（AR）在隐睾症小鼠睾丸管周细胞中的表达水平及对隐睾症导致生精功能障碍的理论意义。
方法
2
30只5周龄雄性ICR小鼠采用随机数字表法随机分配至6组中，随机抽取3组15只小鼠进行手术诱导隐睾，其余3组为作为对照组进行假手术处理。分别于4 d、7 d、14 d后取各组睾丸组织，然后测量睾丸体积、观察睾丸组织病理，提取各组睾丸管周细胞后利用免疫荧光、Real-Time PCR和蛋白质印记法检测AR和GDNF的mRNA和蛋白的表达。
结果
2
对照组4 d、7 d、14 d小鼠的睾丸体积分别为（125.58±19.22）mm
3
、（123.45±20.12）mm
3
、（140.09±13.62）mm
3
；睾丸各级生精细胞排列整齐、层次清楚，可见较多精子细胞，生精小管周围管周细胞形态规则，呈梭形围绕小管周围，细胞厚度均一；ARmRNA的表达量分别为1.00±0.05、1.06±0.07、1.19±0.13GDNFmRNA的表达量分别为1.00±0.04、1.09±0.05、1.10±0.07；AR蛋白的表达量分别为1.01±0.01、0.79±0.02、1.01±0.04；GDNF蛋白的浓度分别为（18.68±0.43）pg/mL、（14.39±0.36）pg/mL、（16.88±0.37）pg/mL。隐睾组4 d、7 d、14 d小鼠的睾丸体积分别为（115.64±3.91）mm
3
、（69.51±14.97）mm
3
、（44.86±5.56）mm
3
；睾丸各级生精细胞排列紊乱、层次不清、结构破坏，曲细精管周围管周细胞萎缩、弯曲断裂；ARmRNA的表达量分别为0.76±0.06、0.53±0.04、0.29±0.02；GDNFmRNA的表达量分别为0.72±0.05、0.42±0.02、0.30±0.03；AR蛋白的表达量分别为0.54±0.02、0.98±0.04、0.31±0.01；GDNF蛋白的浓度分别为（8.50±0.34）pg/mL、（17.44±0.32）pg/mL、（6.83±0.34）pg/mL。上述指标与对照组相比，除了4 d的睾丸体积差异无统计学意义（
P
>
0.05），其他均有统计学差异（
P
<
0.05）。对照组中3个时间点的睾丸体积、AR和GDNF的mRNA、蛋白表达量的差异无统计学意义（
P
>
0.05），隐睾组3个时间点的睾丸体积、AR和GDNF的mRNA、蛋白表达量呈逐渐下降趋势且各组之间的差异均有统计学意义（
P
<
0.05）。
结论
2
在手术诱导隐睾小鼠中，睾丸管周细胞的AR和GDNF的表达水平随着诱导时间的延长呈现显著下降。AR和GDNF在隐睾症介导睾丸管周细胞功能的损伤中有重要作用。本研究为阐明隐睾症导致生精功能障碍的机制研究提供理论基础。
Abstract
Objective
2
To investigate the expression of glial cell line-derived neurotrophic factor （GDNF） and androgen receptor （AR） in testicular peritubular cells （TPCs） of cryptorchidism mouse models and explore the theoretical significance of cryptorchidism-induced spermatogenesis dysfunction.
Methods
2
A total of 30 five-week-old male ICR rats were divided randomly by using random number table method into 6 groups. Cryptorchidism was surgically induced in 3 randomly selected groups and the other 3 groups underwent sham surgery as the control groups. On days 4， 7 and 14 after surgery， we harvested the mice testes of the 3 groups and their corresponding control groups， then measured the testicular volumes， analyzed the testicular histopathology and detected the mRNA and protein expression levels of AR and GDNF in TPCs by immunofluorescence， real-time PCR and Western blot.
Results
2
In normal control groups， on days 4， 7 and 14 after surgery， the testicular volumes were （125.58±19.22） mm
3
，（123.45±20.12） mm
3
， （140.09±13.62） mm
3
， respectively. Clear layers of spermatogenic cells were well arranged and abundant sperm cells were found. Peritubular cells were morphologically homogeneous， with slim-spindle appearance and normal cell thickness. The mRNA expression levels of AR were 1.00±0.05， 1.06±0.07 and 1.19±0.13； GDNF mRNA 1.00±0.04， 1.09±0.05， and 1.10±0.07. The protein expression levels of AR were 1.01±0.01， 0.79±0.02 and 1.01±0.04； GDNF protein （18.68±0.43） pg/mL， （14.39±0.36） pg/mL and （16.88±0.37） pg/mL. In cryptorchidism groups， on days 4， 7 and 14 after surgery， the testicular volumes were （115.64±3.91） mm
3
， （69.51±14.97） mm
3
and （44.86±5.56） mm
3
， respectively. Spermatogenic cells were disorganized， seminiferous tubules were disrupted， peritubular cells shrank， bent and fractured. The mRNA expression levels of AR were 0.76±0.06， 0.53±0.04， and 0.29±0.02； GDNF mRNA 0.72±0.05， 0.42±0.02 and 0.30±0.03. The protein expression levels of AR were 0.54±0.02， 0.98±0.04 and 0.31±0.01； GDNF protein （8.50±0.34） pg/mL， （17.44±0.32） pg/mL and （6.83±0.34） pg/mL. Statistically significant differences （
P
<
0.05） were found in 7-day and 14-day testicular volumes between control and cryptorchidism groups but not in the 4-day testicular volume （
P
>
0.05）. Testicular volumes， AR and GDNF mRNA and protein expression in control groups had no statistically significant difference （
P
>
0.05）， while those in cryptorchidism groups showed a trend of gradual decline in the amount and the differences between groups were statistically significant （
P
<
0.05）.
Conclusions
2
In surgery-induced cryptorchidism mice， after the induction， the expression of AR and GDNF in TPCs showed a gradual decrease over time. AR and GDNF play a major role in mediating the TPCs damage in cryptorchidism. This study provides a theoretical basis for mechanism researches of cryptorchidism-induced spermatogenesis dysfunction.
关键词
隐睾症管周细胞胶质细胞系衍生神经营养因子雄激素受体
Keywords
cryptorchidismtesticular peritubular cells （TPCs）glial cell line-derived neurotrophic factor （GDNF）androgen receptor （AR）
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