Department of Endocrinology, Institute of Zoology and Biomedical Research, Jagiellonian University, Krakow, Poland.
Department of Endocrinology, Institute of Zoology and Biomedical Research, Jagiellonian University, Krakow, Poland.
Chemosphere. 2018 May;199:182-190. doi: 10.1016/j.chemosphere.2018.02.011. Epub 2018 Feb 5.
The study was designed to examine the effects of model plastic derived compounds, bisphenol A (BPA) and dibutyl phthalate (DBP), on juxtacrine communication in adult rat testis, by evaluating the expression of Notch pathway components. Testicular explant were exposed in vitro to BPA (5 × 10 M, 2.5 × 10 M, 5 × 10 M) or DBP (10 M, 10 M, 10 M) for 24 h. To determine the expression of Notch1, Dll4, Hey1, Hes1 and Hey5 real-time RT-PCR was used. Protein levels and localization of NOTCH1 receptor, its ligand DLL4 as well as HEY1, HES1 and HEY5 factors were detected by western blot analysis and immunohistochemistry, respectively. Upregulation of Notch1, Dll4 and Hey1 at the mRNA and protein level was demonstrated in testis explants after BPA and DBP treatment (p < 0.05; p < 0.01; p < 0.001). Hes5 expression decreased after BPA (p < 0.05; p < 0.01; p < 0.001), whereas Hes1 expression was not altered by either BPA or DBP. Tested chemicals altered immunoexpression of activated NOTCH1, DLL4, HEY1 and HES5 both in seminiferous epithelium and interstitial tissue, exerting differential effects on particular cell types. In conclusion, BPA and DBP affect Notch signaling pathway in rat testis, which indicates that juxtacrine communication is a potential target for the action of plastic derived compounds in male gonad.
这项研究旨在通过评估 Notch 通路成分的表达,研究模型塑料衍生化合物双酚 A(BPA)和邻苯二甲酸二丁酯(DBP)对成年大鼠睾丸旁细胞通讯的影响。将睾丸组织外植体在体外暴露于 BPA(5×10⁻⁶ M、2.5×10⁻⁶ M、5×10⁻⁶ M)或 DBP(10 M、10 M、10 M)中 24 小时。使用实时 RT-PCR 测定 Notch1、Dll4、Hey1、Hes1 和 Hey5 的表达。通过 Western blot 分析和免疫组织化学分别检测 NOTCH1 受体、其配体 DLL4 以及 HEY1、HES1 和 HEY5 因子的蛋白水平和定位。BPA 和 DBP 处理后,睾丸外植体中 Notch1、Dll4 和 Hey1 的 mRNA 和蛋白水平均上调(p<0.05;p<0.01;p<0.001)。BPA 后 Hes5 表达降低(p<0.05;p<0.01;p<0.001),而 BPA 或 DBP 均未改变 Hes1 表达。测试的化学物质改变了激活的 NOTCH1、DLL4、HEY1 和 HES5 在生精上皮和间质组织中的免疫表达,对特定细胞类型产生不同的影响。总之,BPA 和 DBP 影响大鼠睾丸中的 Notch 信号通路,表明旁分泌通讯是塑料衍生化合物在男性性腺中作用的潜在靶点。