Histopathological changes and pathological basis of the cryptorchidtestis in rats by maternal exposure to flutamide

Peng Jinpu; Zhang Deying; Cao Xining; Zhou Yue; Shen Lianju; Long Chunlan; He Dawei; Lin Tao; Wei Guanghui

doi:10.13406/j.cnki.cyxb.001153

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Histopathological changes and pathological basis of the cryptorchidtestis in rats by maternal exposure to flutamide

更新时间：2024-01-17

- Histopathological changes and pathological basis of the cryptorchidtestis in rats by maternal exposure to flutamide
- Journal of Chongqing Medical University Vol. 42, Issue 8, Pages: 953-958(2017)
- 作者机构：
  
  重庆医科大学附属儿童医院泌尿外科儿童发育疾病研究教育部重点实验室儿科学重庆市重点实验室重庆市儿童发育重大疾病诊治与预防国际科技合作基地
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.001153
  CLC： R-332;R726.9
- Published：2017，
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[1]彭金普,张德迎,曹希宁,周玥,沈炼桔,龙春兰,何大维,林涛,魏光辉.氟他胺致大鼠隐睾模型的睾丸组织形态改变及其病理基础[J].重庆医科大学学报,2017,42(08):953-958.

Peng Jinpu, Zhang Deying, Cao Xining, et al. Histopathological changes and pathological basis of the cryptorchidtestis in rats by maternal exposure to flutamide. [J]. Journal of Chongqing Medical University 42(8):953-958(2017)
[1]彭金普,张德迎,曹希宁,周玥,沈炼桔,龙春兰,何大维,林涛,魏光辉.氟他胺致大鼠隐睾模型的睾丸组织形态改变及其病理基础[J].重庆医科大学学报,2017,42(08):953-958. DOI： 10.13406/j.cnki.cyxb.001153.

Peng Jinpu, Zhang Deying, Cao Xining, et al. Histopathological changes and pathological basis of the cryptorchidtestis in rats by maternal exposure to flutamide. [J]. Journal of Chongqing Medical University 42(8):953-958(2017) DOI： 10.13406/j.cnki.cyxb.001153.

摘要

目的:研究抗雄激素受体氟他胺（Flutamide

Flu）孕中、晚期诱导子代雄鼠发生隐睾

及其病理组织学损害

探讨其不育的病理学基础

为临床治疗隐睾所致非梗阻性不育提供依据。方法:将20只SD（sprague dawley）孕鼠随机分为Flu隐睾组（n=10）、正常对照组（n=10）

Flu隐睾组于妊娠12

1 d给予Flu 25 mg/（kg·d）皮下注射

正常对照组不予任何处理

取各组出生后第60天（postnatal day 60

PND60）的雄性SD大鼠睾丸组织（Flu隐睾组只取隐睾睾丸）

HE染色观察睾丸组织形态学的差异

透射电镜观察睾丸支持细胞间的紧密连接结构

TUNEL凋亡染色检测生精细胞凋亡情况

取附睾尾进行精子计数及形态观测

免疫组织化学和Western blot检测睾丸组织中生殖细胞增殖分化指标视黄酸刺激因子8（stimulated by retinoic acid gene 8

Stra8）、联会复合体蛋白3（synaptonemal complex protein 3

SCP3）的表达

采用Q-PCR检测Stra8基因水平。结果:收集正常对照组正常睾丸30只与Flu隐睾组隐睾睾丸22只

HE染色显示隐睾睾丸管腔明显缩窄、生精细胞发育迟缓且排列紊乱、管腔中央无精子形成;TUNEL凋亡检测证实隐睾组有大量生精细胞发生凋亡

精子计数结果为隐睾组（1.99±0.13）×108个/m L

远低于正常对照组[（5.53±0.17）×108个/m L

P=0.000];透射电镜（transmission electron microscope

TEM）可观察到SD大鼠隐睾支持细胞间的紧密连接结构疏松;免疫组织化学显示Stra8在隐睾组织中表达较正常对照组少

SCP 3在正常对照组睾丸生精小管的各级生精细胞胞核中均有表达

而在隐睾组睾丸中表达很弱;Western blot结果显示

Flu隐睾组Stra8蛋白表达量（0.34±0.05）明显低于正常对照组（0.96±0.09）

P=0.002;Flu隐睾组SCP3蛋白表达量（0.39±0.03）也明显低于正常对照组（0.97±0.07）

P=0.001。Q-PCR结果显示

Flu组SD大鼠睾丸组织内Stra8 m RNA的表达（0.765±0.015）较正常对照组（1.00±0.01）低

P=0.01。结论:Flu诱导的SD大鼠隐睾模型中

睾丸呈明显病理形态学改变

支持细胞间紧密连接结构破坏

Stra8及SCP3表达明显下调

生精细胞凋亡增多

精子数量及质量下降

这可能是导致生精细胞发育障碍的重要原因。

Abstract

Objective:To explore Flutamide(Flu)induced cryptorchidism in pregnant rats and its histopathological changes and pathological basis;to provide references for treating cryptorchidism induced non obstructive infertility. Methods:Totally 20 SD pregnant rats were randomly divided into Flu cryptorchidism group(n=10)and normal control group(n=10). Flu was given 25 mg/(kg·d)by subcutaneous injection on pregnancy 12 to 21 days in Flu cryptorchidism group;nothing was given normal control group. When male SD rats were 60 days after birth

testis tissue were collected

and testicular histological morphology was observed by HE staining

Tight junction between Sertoli cells was detected by TEM;TUNEL staining was used to detect the apoptosis of spermatogenic cells;cauda epididymal was achieved for sperm count and morphology analysis;protein expressions of Stra8 and SCP3 were detected by immunohistochemistry and Western blot;m RNA expression of Stra8 was examined through Q-PCR. Results:Thirty normal testis and 22 undescended testis were collected. HE images showed the cryptorchid testis lumen decreased and narrowed significantly;spermatogenic cell retardated and arranged in disorder;no sperm formed in central lumen. The tight junctions of Sertoligerm cells were abnormal in cryptorchid testis. TUNEL apoptosis and sperm count with morphology observation results showed that there are a large number of spermatogenic cells apoptosis;the number of sperm of cryptorchidism group was decreased compared with that in normal group(P=0.000)[(1.99±0.13)×108 cell/m L vs.(5.53±0.17)×108 cell/m L]. The protein expressions of Stra8 and SCP3 were significantly lower in the cryptorchid testicular tissue than in the normal control group. Q-PCR showed that m RNA expression of Stra8 was significantly lower in cryptorchidism group than in the normal group[(0.765±0.015)vs.(1.00±0.01)]

P=0.010. Conclusion:In SD rat cryptorchidism model induced by Flu

there is an obvious pathological change in the testis

the tight junctions between Sertoli cells are destroyed

expression of Stra8 and SCP3 is significantly decreased

which may be very important during the process of spermatogenic impairment in cryptorchidism.

关键词

氟他胺大鼠隐睾睾丸病理改变

Keywords

flutamideratcryptorchidismtestishistopathological change

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