Department of Urology, Shandong Provincial Hospital, Shandong University, Jinan, Shandong, China.
Department of Urology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, China.
Syst Biol Reprod Med. 2024 Dec;70(1):289-298. doi: 10.1080/19396368.2024.2403383. Epub 2024 Oct 3.
Cryptorchidism, a condition where the testis fails to fully descend into the scrotum during development, is associated with elevated environmental temperatures and pressures, leading to male infertility and germ cell tumors. Factors such as oxidative stress and high temperatures contribute to infertility in cryptorchidism. This study aims to explore how external pressure affects Sertoli cells and discover new mechanisms affecting spermatogenesis in cryptorchidism. Sertoli cells were subjected to various pressure levels (0 mmHg, 25 mmHg, 50 mmHg, 100 mmHg) and durations (0 h, 2 h, 4 h) using an enzyme-linked immunosorbent assay (ELISA) to measure androgen binding protein (ABP) and inhibin B (INH B) secretion. Cell morphology changes were observed using immunofluorescence; apoptosis rates were measured with terminal-deoxynucleotidyl transferase mediated nick end labelling (TUNEL) assay and flow cytometry; ultrastructural variations were examined transmission electron microscopy; and the expression of apoptosis-related proteins (, , , and ) was analyzed through immunohistochemistry, real-time polymerase chain reaction (real-time PCR), and western blotting. The results showed that elevated pressure suppressed ABP and INH B secretion from Sertoli cells. Structural changes were observed under pressure, including cytoskeleton loosening and nuclear fragmentation. Apoptosis rates increased with higher pressure levels. Ultrastructural analysis revealed chromatin changes, apoptotic bodies, and mitochondrial alterations. Increased expressions of and were detected, along with elevated levels of and . The inhibitor blocked pressure-induced apoptosis and activation, while the cytochrome C inhibitor did not show the same effect. Our findings suggested that external pressure induces apoptosis of Sertoli cells the / signaling pathway, potentially contributing to male infertility associated with cryptorchidism.
隐睾症是一种睾丸在发育过程中未能完全下降到阴囊的疾病，与环境温度和压力升高有关，导致男性不育和生殖细胞肿瘤。氧化应激和高温等因素导致隐睾症不育。本研究旨在探讨外部压力如何影响支持细胞，并发现影响隐睾症生精的新机制。使用酶联免疫吸附试验（ELISA）使支持细胞承受不同的压力水平（0mmHg、25mmHg、50mmHg、100mmHg）和持续时间（0h、2h、4h），以测量雄激素结合蛋白（ABP）和抑制素 B（INH B）的分泌。使用免疫荧光观察细胞形态变化；使用末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）测定和流式细胞术测量细胞凋亡率；使用透射电子显微镜检查超微结构变化；通过免疫组织化学、实时聚合酶链反应（real-time PCR）和蛋白质印迹分析凋亡相关蛋白（、、、和）的表达。结果表明，升高的压力抑制了支持细胞中 ABP 和 INH B 的分泌。在压力下观察到结构变化，包括细胞骨架松动和核碎片。随着压力水平的升高，细胞凋亡率增加。超微结构分析显示染色质变化、凋亡小体和线粒体改变。检测到 和 的表达增加，同时 和 的水平升高。 抑制剂阻断了压力诱导的凋亡和 激活，而细胞色素 C 抑制剂则没有相同的作用。我们的研究结果表明，外部压力通过 / 信号通路诱导支持细胞凋亡，可能导致与隐睾症相关的男性不育。
