Nabighadim Amirreza, Jafarnezhad-Ansariha Fahimeh, Majidi Zolbin Masoumeh, Daryabari Seyedeh Sima, Fendereski Kiarad, Kajbafzadeh Abdol Mohammad
Pediatric Urology and Regenerative Medicine Research Center, Children's Medical Center, Tehran University of Medical Sciences, Tehran, Iran.
Cent European J Urol. 2021;74(1):99-108. doi: 10.5173/ceju.2020.0175. Epub 2020 Dec 31.
Approximately 15% of couples in the reproductive age are struggling with infertility which, in nearly half of them, is caused by male factors.
The present study comprised of two groups of sixteen C57BL/6 mice; each mouse received either an intraperitoneal injection of 30 mg/kg of an alkylating agent or the same amount of distilled water. Testes were harvested 30 days following the injection. Morphometric analysis of hematoxylin and eosin (H&E) stained slides including mean tubular area, diameter and intratubular particles were performed. Spermatogenesis rate was assessed by spermatogonial markers including promyelocytic leukemia zinc finger protein (PLZF) and neurogenin-3 (NGN3). Moreover, the expression rate of Wilms Tumor-1 (WT-1), A-Kinase Anchoring Protein 4 (AKAP4) and adenosine deaminase domain containing 1 (ADAD1) genes were evaluated via real-time polymerase chain reaction (RT-PCR).
The body weight gradually increased in both groups after a period of 30 days, however, the increase was significantly (p-value = 0.023) lower in the chemically treated group. All the morphometric parameters were considerably decreased in the azoospermic mice. Also, promyelocytic leukemia zinc finger protein and neurogenin-3 expression dramatically declined (p-value <0.001 for both markers). In comparison with the negative control group, the expression rates of A-Kinase Anchoring Protein 4 and adenosine deaminase domain containing 1, two genes participating in the sperm structure, were remarkably reduced in the intervention group (p-value <0.001); however, our investigations demonstrated that the azoospermia model could induce a 5-fold upregulation in Wilms Tumor-1 gene expression.
Development of an azoospermia model can upregulate Wilms Tumor-1 gene expression in a higher rate after 30 days; however, expression of the testis-specific genes, A-Kinase Anchoring Protein 4 and adenosine deaminase domain containing 1, decreased after the intervention. To the best of our knowledge, this upregulation could be related to spermatogenesis recovery after the follow-up period.
约15%的育龄夫妇正面临不孕问题，其中近一半是由男性因素导致的。
本研究包括两组，每组16只C57BL/6小鼠；每只小鼠腹腔注射30mg/kg的烷化剂或等量的蒸馏水。注射后30天采集睾丸。对苏木精和伊红（H&E）染色的切片进行形态计量分析，包括平均管腔面积、直径和管腔内颗粒。通过精原细胞标志物，包括早幼粒细胞白血病锌指蛋白（PLZF）和神经生成素-3（NGN3）评估精子发生率。此外，通过实时聚合酶链反应（RT-PCR）评估威尔姆斯瘤-1（WT-1）、A激酶锚定蛋白4（AKAP4）和含腺苷脱氨酶结构域1（ADAD1）基因的表达率。
30天后两组体重均逐渐增加，但化学处理组的增加显著较低（p值=0.023）。无精子症小鼠的所有形态计量参数均显著降低。此外，早幼粒细胞白血病锌指蛋白和神经生成素-3的表达显著下降（两种标志物的p值均<0.001）。与阴性对照组相比，参与精子结构的两个基因A激酶锚定蛋白4和含腺苷脱氨酶结构域1在干预组中的表达率显著降低（p值<0.001）；然而，我们的研究表明，无精子症模型可使威尔姆斯瘤-1基因表达上调5倍。
无精子症模型在30天后可使威尔姆斯瘤-1基因表达以更高的速率上调；然而，干预后睾丸特异性基因A激酶锚定蛋白4和含腺苷脱氨酶结构域1的表达降低。据我们所知，这种上调可能与随访期后精子发生的恢复有关。
