维生素D对完整大鼠睾丸组织结构及精子发生形态学指标的影响

I. Marakhovskyi, Y. Laryanovska, Yevheniia Korenieva, N. Smolienko, E. Chystiakova, І. О. Belkina, N. Velychko, K. Misiura, V. Bondarenko
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引用次数: 0

摘要

目前的问题之一是研究维生素D对人体的影响,而在男性中,它的作用与雄激素缺乏和生育能力低下的发病机制密切相关。需要特别注意确定胆钙化醇(D3)是否对完整个体的性腺和精子发生有负面影响,因为维生素D治疗用于有或没有维生素D缺乏症的生殖障碍。本研究旨在探讨维生素D对成年完整雄性大鼠性腺组织结构及精子发生形态学参数的影响。这项研究是在性活跃的成年雄性Wistar大鼠身上进行的。维生素D3分别以1000iu, 4000iu和10000iu的剂量口服。这些溶液是用种子油配制的。对照组是完整的大鼠。在精子发生和附睾精子成熟期间给予维生素D及其溶剂,之后测定睾丸的结构组织。性腺样品固定在10%福尔马林溶液中,留在增加强度的醇中,并包埋在石蜡中。除了显微镜观察外,还对苏木精和伊红染色的性腺切片进行了精子发生过程的形态计量学评估。显微制剂采用Granum l30(03)光镜检查,显微图像采用Granum DSM 310数码摄像机拍摄。照片在奔腾2.4 GHz计算机上使用俯视图进行处理。数据结果在标准软件包“Statistica 6.0”中进行统计处理,采用Student's t检验,采用单向方差分析的非参数模拟- Kruskal-Wallis准则。在大鼠睾丸切片中,精管呈横向或斜向分布,卵圆形或圆形。小管直径正常,管膜、蛋白、血管膜均正常。基底部包含生发上皮中最年轻的细胞-精原细胞。细胞具有明显的功能性活动。完整大鼠精子发生的形态计量学特征符合生理规范。在精子发生和附睾成熟精子的整个过程中引入溶剂对睾丸的组织结构没有影响。睾丸叶充满同心或扁平的精管剖面,它们彼此足够接近。小管的直径正常,小管的固有膜,以及蛋白质和血管膜与完整动物的一致。小管中可见3-4代不同发育阶段的生精细胞。然而,很少有小管出现局灶性生殖上皮破坏和小管腔内生殖细胞脱落。在服用所有剂量的维生素D后,没有观察到精管微观结构的显著变化。不仅精子发生,而且精子发生在大鼠的不同小管中——从精子到精子的细胞转化阶段。注射不同剂量维生素D3的大鼠精子发生过程的形态计量参数与未注射维生素D3的大鼠无差异。因此,使用这些剂量的维生素D显示其对12期减数分裂的精原细胞和小管数量的影响是安全的。在雄性大鼠体内使用胆酸酚68天后,其精子生成指数保持在未使用该物质的动物的水平。在完整的成年雄性大鼠中使用维生素D3不会对睾丸的组织学结构产生不利影响。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Influence of vitamin D on the histostructure of the testis and morphometric indications of spermatogenesis of intact rats
One of the current problems is the study of the effects of vitamin D on the body, and in men its action is closely related to the pathogenesis of androgen deficiency and hypofertility. Particular attention needs to be paid to determining whether cholecalciferol (D3) has a negative effect on the gonads and spermatogenesis of intact individuals, as vitamin D therapy is used in reproductive disorders with or without vitamin D deficiency. The aim of the study was to determine the effect of vitamin D on the histological structure of gonads and morphometric parameters of spermatogenesis of adult intact male rats. The studies were performed on adult sexually active male Wistar rats. Vitamin D3 was administered orally in doses of 1000 IU, 4000 IU and 10000 IU. The solutions were made on seed oil. The control was intact rats. Vitamin D and its solvent were administered throughout the period of spermatogenesis and the time of maturation of sperm in the epididymis, after which the structural organization of the testes was determined. Gonadal samples were fixed in 10 % formalin solution, leave in alcohols of increasing strength, and embedded in paraffin. In addition to survey microscopy, morphometric evaluation of the process of spermatogenesis was performed on sections of gonads stained with hematoxylin and eosin. Micropreparations were examined using a Granum L 30 (03) light microscope, and microscopic images were taken with a Granum DSM 310 digital video camera. Photographs were processed on a Pentium 2.4 GHz computer using Toup View. Statistical processing of data results was performed in the standard software package "Statistica 6.0" using Student's t-test and using a non-parametric analogue of one-way analysis of variance – Kruskal-Wallis criterion. In rat testicular sections, seminiferous tubules were located transversely or obliquely and were oval or round in shape. The diameter of the tubules is normal, the tubular membrane, as well as the protein and vascular membranes were normal. The basal department contains the youngest cells of the germinal epithelium – spermatogonia. Cells have a pronounced functional activity. Morphometric characteristics of spermatogenesis of intact rats corresponded to the physiological norm. The introduction of the solvent throughout the period of spermatogenesis and maturation of mature sperm in the epididymis did not affect the histoarchitectonics of the testicles. The testicular lobes are filled with concentric or flattened profiles of sections of the seminal tubules, which are close enough to each other. The diameter of the tubules is normal, the intrinsic membrane of the tubules, as well as the protein and vascular membranes corresponded to those in intact animals. 3-4 generations of spermatogenic cells, which were at different stages of development, can be seen in the tubules. However, few tubules with focal destruction of the germinal epithelium and exfoliation of germ cells in the lumen of the tubules have been observed. No significant changes in the microstructure of the seminal tubules were observed after administration of vitamin D at all doses studied. Not only spermatogenesis but also spermiogenesis is clearly traced in different tubules of rats – stages of cellular transformations from spermatid to sperm. Morphometric parameters of the process of spermatogenesis of rats receiving different doses of vitamin D3 do not differ from those of intact rats. Thus, the use of vitamin D in these doses revealed the safety of its effect on the number of spermatogonia and tubules with stage 12 meiosis. When cholecaciferol was used in male rats for 68 days, the spermatogenesis index remained at the level of animals that did not receive the test substance. The use of vitamin D3 in intact adult male rats does not adversely affect the histological structure of the testes.
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