Lv Dongliang, Zhao Mengjie, Ni Jie, Liu Weidong, Ren Yijie, Zhu Dawei, Hu Jianhong
Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China.
Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China.
Theriogenology. 2023 Jan 1;195:138-148. doi: 10.1016/j.theriogenology.2022.10.017. Epub 2022 Oct 18.
Damage to Sertoli cell junction proteins caused by inflammation can lead to male infertility. Nerve growth factor (NGF) plays an important role in reproductive and inflammatory disease; however, whether and how NGF regulates Sertoli cell function remains unclear. Here, we aimed to assess the effect of NGF on the growth of Sertoli cells isolated from the testes of dairy goats and evaluate if NGF has a protective effect on these cells. We confirmed that Sertoli cell viability, proliferation, and ATP content increased following NGF treatment. In addition, qPCR results suggested that Sertoli cell apoptosis was inhibited after NGF treatment. To investigate the protective effect of NGF on Sertoli cells under pathological inflammatory conditions, LPS was used to induce inflammatory response in Sertoli cells. Post-treatment, the entangled filamentous pseudopodia of the cells loosened and no longer spanned adjacent cells. The expression of several junction proteins (ZO-1, occludin, CX-43, β-catenin, and N-cadherin), which was down-regulated after inflammatory response induction, was restored following NGF treatment. LPS-induced changes in cytotoxicity and transepithelial electrical resistance were reversed and the intercellular connections became tighter after NGF treatment. We further demonstrated that NGF prevented the inflammatory response of Sertoli cells via the PI3K/AKT/NFκB signaling pathway, similar to the effect of the PI3K-inhibitor, LY294002, which is modified by the PI3K activator, 740Y-P. These results provide insights for devising strategies for protecting the male reproductive system and curing or preventing associated pathological conditions.
炎症导致的支持细胞连接蛋白损伤可导致男性不育。神经生长因子(NGF)在生殖和炎症性疾病中起重要作用;然而,NGF是否以及如何调节支持细胞功能仍不清楚。在此,我们旨在评估NGF对从奶山羊睾丸分离的支持细胞生长的影响,并评估NGF是否对这些细胞具有保护作用。我们证实,NGF处理后支持细胞的活力、增殖和ATP含量增加。此外,qPCR结果表明,NGF处理后支持细胞凋亡受到抑制。为了研究NGF在病理性炎症条件下对支持细胞的保护作用,使用脂多糖(LPS)诱导支持细胞产生炎症反应。处理后,细胞缠结的丝状伪足松开,不再跨越相邻细胞。几种连接蛋白(ZO-1、闭合蛋白、CX-43、β-连环蛋白和N-钙黏蛋白)的表达在炎症反应诱导后下调,在NGF处理后恢复。LPS诱导的细胞毒性和跨上皮电阻变化被逆转,NGF处理后细胞间连接变得更紧密。我们进一步证明,NGF通过PI3K/AKT/NFκB信号通路阻止支持细胞的炎症反应,类似于PI3K抑制剂LY294002的作用,而LY294002的作用被PI3K激活剂740Y-P修饰。这些结果为制定保护男性生殖系统以及治疗或预防相关病理状况的策略提供了思路。