Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Urogenital Stem Cell Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Andrologia. 2021 Aug;53(7):e14095. doi: 10.1111/and.14095. Epub 2021 May 7.
The current study aims to develop a validated animal model to predict successful spermatogenesis retrieval in azoospermia and oligospermia men. Thirty-two mice were equally divided into 4 groups: control, scrotal hyperthermia (15 times), scrotal hyperthermia group (10 times), scrotal hyperthermia group (5 times). In the scrotal hyperthermia groups, their scrotum exposed to water at a temperature of 43°C for 20 min every other day. Then, the mice were euthanised and sperm samples were collected for sperm parameters analysis, and blood samples were obtained for hormonal assay. The testis samples were taken for histopathology experiments, immunofluorescence staining and Western blot in order to examine the protein expression together with RNA extraction in order to examine the gene expression of germ cell markers. The results of sperm analysis and histopathology of testicular tissue as well as the results of gene expression and Western blot showed that hyperthermia can significantly impair spermatogenesis. In conclusion, we have developed a novel model of azoospermia and oligospermia in mouse, which uses a high temperature to suppress spermatogenesis process through demolition of germ cells subsequent cell cycle arrest and apoptosis. The model will contribute to understanding azoospermia in human, oligospermia pathophysiology and the development of treatment.
本研究旨在建立一种经过验证的动物模型，以预测无精子症和少精子症男性成功获取精子的情况。32 只小鼠被平均分为 4 组：对照组、阴囊高温（15 次）组、阴囊高温（10 次）组、阴囊高温（5 次）组。在阴囊高温组中，每隔一天将其阴囊暴露于 43°C 的水中 20 分钟。然后处死小鼠，收集精子样本进行精子参数分析，并采集血液样本进行激素测定。采集睾丸样本进行组织病理学实验、免疫荧光染色和 Western blot，以检查蛋白表达情况，并提取 RNA 以检查生殖细胞标志物的基因表达情况。精子分析和睾丸组织的组织病理学结果以及基因表达和 Western blot 的结果表明，高温可显著损害精子发生。总之，我们已经在小鼠中建立了一种新的无精子症和少精子症模型，该模型通过破坏生殖细胞来抑制精子发生过程，随后导致细胞周期停滞和细胞凋亡。该模型将有助于了解人类无精子症、少精子症的病理生理学以及治疗方法的开发。
