de Freitas André Teves Aquino Gonçalves, Ribeiro Mariana Antunes, Pinho Cristiane Figueiredo, Peixoto André Rebelo, Domeniconi Raquel Fantin, Scarano Wellerson R
Graduate Program in General and Applied Biology, Institute of Biosciences, UNESP, Botucatu, SP, Brazil.
Department of Anatomy, Institute of Biosciences, UNESP, Botucatu, SP, Brazil.
Toxicol In Vitro. 2016 Aug;34:1-7. doi: 10.1016/j.tiv.2016.02.017. Epub 2016 Feb 26.
The blood-testis barrier (BTB) is responsible for providing a protected environment and coordinating the spermatogenesis. Endocrine disruptors (EDs) might lead to infertility, interfering in the BTB structure and modulation. This study aimed to correlate the actions of two EDs, monobutyl phthalate (MBP) and bisphenol A (BPA) in different periods of exposure, in a low toxicity dose to the human Sertoli cells (HSeC) and its effects on the proteins of the BTB and regulatory proteins involved in its modulation. HSeC cells were exposed to MBP (10μM) and BPA (20μM) for 6 and 48h. Western Blot assay indicated that MBP was able to reduce the expression of occludin, ZO-1, N-cadherin and Androgen Receptor (AR), while BPA leads to a reduction of occludin, ZO-1, β-catenin and AR. TGF-β2 and F-actin were not modified. Phalloidin and Hematoxylin and Eosin assay revealed phenotically disruption in Sertoli cells adhesion, without changes in F-actin expression or localization. Our data suggested both EDs present potential for disrupting the structure and maintenance of the human BTB by AR dependent pathway.
血睾屏障（BTB）负责提供一个受保护的环境并协调精子发生过程。内分泌干扰物（EDs）可能会干扰BTB的结构和调节，从而导致不育。本研究旨在关联两种内分泌干扰物邻苯二甲酸单丁酯（MBP）和双酚A（BPA）在不同暴露时期、以低毒性剂量作用于人类支持细胞（HSeC）时的作用，及其对BTB相关蛋白和参与其调节的调控蛋白的影响。将HSeC细胞分别暴露于MBP（10μM）和BPA（20μM）中6小时和48小时。蛋白质免疫印迹分析表明，MBP能够降低闭合蛋白、紧密连接蛋白1（ZO-1）、N-钙黏蛋白和雄激素受体（AR）的表达，而BPA则导致闭合蛋白、ZO-1、β-连环蛋白和AR的表达降低。转化生长因子-β2（TGF-β2）和F-肌动蛋白未发生改变。鬼笔环肽和苏木精-伊红染色分析显示支持细胞黏附出现表型破坏，而F-肌动蛋白的表达或定位没有变化。我们的数据表明，这两种内分泌干扰物都有可能通过雄激素受体依赖途径破坏人类血睾屏障的结构和维持。
