The fungicide tebuconazole (TEB) poses risks to human and animal health via various exposure routes. It induces toxicity in multiple organs and disrupts reproductive health by affecting steroid hormone synthesis and fetal development. In this study, we investigated the impact of TEB on fetal testes using in vitro models, focusing on germ, Sertoli, and Leydig cells, and explored the mechanisms underlying cellular damage. The results revealed significant damage to germ cells and disruption of Leydig cell development. TEB exposure led to a decrease in germ cell numbers, as indicated by histological and immunostaining analyses. TEB induced the up- and down-regulation of the expression of fetal and adult Leydig cell markers, respectively. Additionally, TEB-treated fetal testes exhibited increased expression of oxidative-stress-related genes and proteins. However, co-treatment with the antioxidant N-acetylcysteine mitigated TEB-induced germ cell damage and prevented abnormal Leydig cell development. These findings suggest that administration of antioxidants can prevent the intratesticular damage typically caused by TEB exposure.

Humans are exposed to a multitude of endocrine disruptor chemicals (EDCs) that can interfere with the action of endogenous hormones and the normal development of reproductive organs. Bisphenol A (BPA) is one of the most common EDCs found in the environment. Here, we evaluated BPA toxicity on fetal testes using an in vitro organ culture system. Mouse fetal testes sampled at 15.5 days post coitus were cultured in a medium containing BPA for 5 days. The number of germ cells was reduced by BPA treatment, whereas the number of Sertoli cells was slightly increased by BPA at the highest dose (100 μM). Consistently, BPA treatment reduced the protein and gene expression levels of germ cell markers, but it increased the expression levels of Sertoli cell markers. The expression levels of fetal Leydig cell markers such as Cyp11a1, Thbs2, Cyp17a1, and Pdgf-α were significantly increased, whereas those of adult Leydig cell markers such as Hsd17b3, Ptgds, Sult1e1, Vcam1, and Hsd11b1 were decreased in the testes exposed to BPA. Generally, Notch signaling restricts Leydig cell differentiation from progenitor cells during fetal testis development. The expression levels of Notch1, Notch2, Notch3, Hes1, Ptch1, Jag1, Jag2, c-Myc, Hey1, and Hey2, which are involved in Notch signaling, were markedly higher in BPA-treated fetal testes than in the controls, indicating that BPA interrupts fetal Leydig cell development. BPA also disrupted steroidogenesis in the fetal testis organ culture system. In conclusion, our study showed that BPA inhibits fetal germ cell growth, Leydig cell development, and steroidogenesis.

OBJECTIVE: To study the morphological characteristics and expression of vascular endothelial growth factor (VEGF) in the fetal testes exposed to chronic intrauterine hypoxia during pathological pregnancy in different gestation periods.
METHODS: The testes from 48 male fetuses that had died in the antenatal or early neonatal period in mothers with pathological pregnancy were morphologically evaluated.
RESULTS: Chronic intrauterine hypoxia was shown to be a powerful damaging factor and leads to delayed gonadal development. Histological examination of testicular tissue showed a significant reduction in the number of tubular cells per vision field, a decrease in tubular diameter and area, with the simultaneously increased area of the stroma and a larger number of vessels. Immunohistochemical study revealed the pronounced cytoplasmic expression of VEGF in testicular tissue in different gestation periods in the spermatogenic epitheliocytes, vessels, Leydig interstitial cells, while the maximal expression of this receptor was observed at 19-25 weeks\' gestation, the degree of expression decreased at 26-29 weeks\' gestation.
CONCLUSIONS: Intrauterine hypoxia has a destabilizing effect on the processes of proliferation and differentiation of the spermatogenic epithelium, interstitial endocrinocytes, activates the processes of angiogenesis and the growth of connective tissue. All this can involve not only gonadal dysgenesis, but also future reproductive dysfunction. Hypoxia stimulates the expression of VEGF, whose receptors are present in almost all testicular cell populations. It can be assumed that VEGF can act as a paracrine regulator of Leydig cell activity, also as an inducer of angiogenesis, and thus play a certain role in the development of male fertility.
Цель исследования - изучить морфологические особенности и экспрессию васкулоэндотелиального фактора роста в яичках плодов, подвергшихся влиянию хронической внутриутробной гипоксии при патологическом течении беременности на разных сроках гестации. Материал и методы. Проведена морфологическая оценка яичек 48 плодов, умерших в антенатальный или ранний неонатальный период у матерей с патологическим течением беременности. Результаты. Установлено, что хроническая внутриутробная гипоксия является мощным повреждающим фактором и приводит к задержке развития гонад. При гистологическом исследовании ткани яичек отмечалось достоверное снижение количества клеток в канальцах в поле зрения, уменьшение диаметра и площади канальцев при одновременном увеличении площади стромы и количества сосудов. При иммуногистохимическом исследовании наблюдалась выраженная цитоплазматическая экспрессия VEGF в ткани яичек в разные сроки гестации в клетках сперматогенного эпителия, сосудах, интерстициальных клетках Лейдига, причем максимальная экспрессия данного рецептора отмечена в сроки 19-25 нед гестации, при сроке 26-29 нед степень выраженности экспрессии уменьшалась. Заключение. Внутриутробная гипоксия оказывает дестабилизирующее влияние на процессы пролиферации и дифференцировки сперматогенного эпителия, интерстициальных эндокриноцитов, активирует процессы ангиогенеза и рост соединительной ткани. Все это может повлечь не только аномальное формирование гонад, но и в дальнейшем нарушение репродуктивной функции. Гипоксия стимулирует экспрессию сосудистого эндотелиального фактора роста, рецепторы которого (VEGFR) присутствуют почти во всех популяциях тестикулярных клеток. Можно предположить, что VEGF, помимо индуктора ангиогенеза, может выступать в качестве паракринного регулятора активности клеток Лейдига и тем самым играть определенную роль в становлении мужской фертильности.