Florea Adrian, Puică Constantin, Hamed Sami, Tilinca Mariana, Matei Horea
Department of Cell and Molecular Biology, Faculty of Medicine, "Iuliu Haţieganu" University of Medicine and Pharmacy, 6 Louis Pasteur St., 400349 Cluj-Napoca, Romania.
Department of Physiology, Institute of Biological Research, 48 Gheorgh Bilaşcu St., 400015 Cluj-Napoca, Romania.
Micron. 2017 Nov;102:1-14. doi: 10.1016/j.micron.2017.07.012. Epub 2017 Aug 16.
We tested here the ability of bee venom (BV) to interfere with spermatogenesis in rats in two experimental conditions. The histopathological changes were assessed with brightfield microscopy using a novel staining technique, based on methylene blue, orange G and ponceau xylidine. Transmission electron microscopy was also used to identify fine subcellular changes. BV injection for 30days in daily doses of 700μg BV/kg resulted in reducing testicular weight, along with significant larger diameters of seminiferous tubules and reduced number of Sertoli cells (SCs). SCs were vacuolated, detached from the basement membrane, many necrosed, leading to the basement membrane denudation. Germ cells layers were separated by empty spaces conferring a rarefied aspect to the tissue, and spermatids were detached into lumen. Thus, the seminiferous epithelium was significantly thinned. Many Leydig cells (LCs) were in a necrotic state, with disrupted plasma membrane and without smooth endoplasmic reticulum. The acute treatment with a single LD50 of 62mgBV/kg, was followed by focal disruptions of the basement membrane and localized areas of necrosis, mainly affecting the SCs. Most of the observed SCs as well as some spermatogonia were highly vacuoled, empty spaces being observed within the epithelium. The SCs count was significantly decreased. Spermatids had also the tendency of separation from the SCs, and the significant larger diameter of the tubules found was associated with a thicker epithelium. Many LCs were necrosed, with disrupted plasma membrane, swollen mitochondria, no endoplasmic reticulum and implicitly showing rarefied cytoplasm. We concluded that BV was a testicular toxicant affecting both the LCs and the seminiferous tubules. The SCs cells represented the primary target site of BV whose effects were next extended upon the germ cells. In all cells, BV triggered unspecific degenerative changes that could impaire spermatogenesis. The present study also proposes an alternative staining technique very useful in assessing the histopathological aspects of spermatogenesis.
我们在此测试了蜂毒（BV）在两种实验条件下干扰大鼠精子发生的能力。使用基于亚甲蓝、橘黄G和丽春红二甲苯黄的新型染色技术，通过明场显微镜评估组织病理学变化。还使用透射电子显微镜来识别细微的亚细胞变化。以每日700μg BV/kg的剂量注射BV 30天，导致睾丸重量减轻，同时生精小管直径显著增大，支持细胞（SCs）数量减少。SCs出现空泡化，从基底膜脱离，许多发生坏死，导致基底膜剥脱。生殖细胞层被空隙分隔，使组织呈现稀疏外观，精子细胞脱落在管腔中。因此，生精上皮显著变薄。许多睾丸间质细胞（LCs）处于坏死状态，细胞膜破裂，没有滑面内质网。单次注射62mg BV/kg的半数致死剂量进行急性处理后，基底膜出现局灶性破坏和局部坏死区域，主要影响SCs。观察到的大多数SCs以及一些精原细胞高度空泡化，上皮内可见空隙。SCs计数显著减少。精子细胞也有与SCs分离的趋势，发现的小管直径显著增大与上皮增厚有关。许多LCs坏死，细胞膜破裂，线粒体肿胀，没有内质网，细胞质明显稀疏。我们得出结论，BV是一种睾丸毒物，会影响LCs和生精小管。SCs细胞是BV的主要靶位点，其作用随后扩展到生殖细胞。在所有细胞中，BV引发非特异性退行性变化，可能损害精子发生。本研究还提出了一种替代染色技术，对评估精子发生的组织病理学方面非常有用。
