Department of Urology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, China.
Department of Urology, Shanghai General Hospital Affiliated to Nanjing Medical University, Shanghai 200080, China.
Toxicology. 2022 May 30;474:153213. doi: 10.1016/j.tox.2022.153213. Epub 2022 May 20.
Dibutyl phthalate (DBP) is an endocrine disruptor, which causes male reproductive dysfunction in rodents. Previous researches demonstrated that DBP exposure impaired spermatogenesis, however, the molecular mechanism is largely uncovered. In this study, we demonstrated that prenatal exposure to DBP increased receptor activator of nuclear factor-κB ligand (RANKL) expression in seminiferous tubules, especially in Sertoli cells. Western blot and immunofluorescence assays showed that DBP induced up-regulation of RANKL expression in Sertoli cells. Furthermore, experimental data showed that DBP increased COX-2 and p-p65 expression in Sertoli cells and depleting COX-2 and p-p65 by specific inhibitor NS3-98 and BAY117082 could partially abolish DBP induced up-regulation of RANKL. Moreover, the content of RANKL in Sertoli cells was significantly increased after DBP exposure by conducting enzyme linked immunosorbent assay (ELISA), which promoted spermatogonial stem cells (C18-4 cells) apoptosis in a paracrine manner. Together, our data demonstrated that a novel mechanism for DBP induced impairment of spermatogenesis by activating NF-κB/COX-2/RANKL signaling in Sertoli cells, and provided a diagnostic and therapeutic target.
邻苯二甲酸二丁酯（DBP）是一种内分泌干扰物，可导致啮齿动物雄性生殖功能障碍。先前的研究表明，DBP 暴露会损害精子发生，但分子机制在很大程度上尚未被揭示。在这项研究中，我们证明了产前暴露于 DBP 会增加生精小管中核因子-κB 受体激活剂配体（RANKL）的表达，尤其是在支持细胞中。Western blot 和免疫荧光分析表明，DBP 诱导支持细胞中 RANKL 的表达上调。此外，实验数据表明，DBP 增加了支持细胞中 COX-2 和 p-p65 的表达，并且通过特异性抑制剂 NS3-98 和 BAY117082 耗尽 COX-2 和 p-p65 可以部分消除 DBP 诱导的 RANKL 上调。此外，通过酶联免疫吸附测定（ELISA）进行 DBP 暴露后，支持细胞中 RANKL 的含量显着增加，这以旁分泌方式促进精原干细胞（C18-4 细胞）凋亡。总之，我们的数据表明，DBP 通过激活支持细胞中的 NF-κB/COX-2/RANKL 信号通路导致精子发生受损的一种新机制，并为诊断和治疗提供了一个靶点。
