Abstract：
Bisphenol A (BPA) is widely exposed in populations worldwide and has negative effects on spermatogenesis both in animals and humans. The homeostasis of the actin cytoskeleton in the spermatogenic epithelium is crucial for spermatogenesis. Actin cytoskeleton destruction in the seminiferous epithelium is one of the important reasons for BPA-induced spermatogenesis disorder. However, the underlying molecular mechanisms remain largely unexplored. Herein, we explored the role and mechanism of Rsad2, an interferon-stimulated gene in BPA-induced actin cytoskeleton disorder in mouse GC-2 spermatocyte cell lines. After BPA exposure, the actin cytoskeleton was dramatically disrupted and the cell morphology was markedly altered accompanied by a significant increase in Rsad2 expression both in mRNA and protein levels in GC-2 cells. Furthermore, the phalloidin intensities and cell morphology were restored obviously when interfering with the expression of Rsad2 in BPA-treated GC-2 cells. In addition, we observed a significant decrease in intracellular ATP levels after BPA treatment, while the ATP level was obviously upregulated when knocking down the expression of Rsad2 in BPA-treated cells compared to cells treated with BPA alone. Moreover, Rsad2 relocated to mitochondria after BPA exposure in GC-2 cells. BPA promoted Rsad2 expression by activating type I IFN-signaling in GC-2 cells. In summary, Rsad2 mediated BPA-induced actin cytoskeletal disruption in GC-2 cells, which provided data to reveal the mechanism of BPA-induced male reproductive toxicity.
摘要：
双酚A（BPA）在世界范围内广泛暴露于人群中，对动物和人类的精子发生都有负面影响。生精上皮中肌动蛋白细胞骨架的稳态对于精子发生至关重要。生精上皮中肌动蛋白细胞骨架的破坏是BPA诱导的精子发生障碍的重要原因之一。然而,潜在的分子机制在很大程度上仍未被探索。在这里，我们探讨了干扰素刺激基因Rsad2在BPA诱导的小鼠GC-2精母细胞肌动蛋白细胞骨架紊乱中的作用和机制。BPA暴露后,在GC-2细胞中,肌动蛋白细胞骨架被显著破坏,细胞形态发生显著改变,同时Rsad2在mRNA和蛋白水平上的表达显著增加。此外，干扰BPA处理的GC-2细胞中Rsad2的表达时，连环肽的强度和细胞形态明显恢复。此外,我们观察到BPA治疗后细胞内ATP水平显着下降，而与单独用BPA处理的细胞相比，当敲除BPA处理的细胞中Rsad2的表达时，ATP水平明显上调。此外，在GC-2细胞中BPA暴露后，Rsad2重新定位到线粒体。BPA通过激活GC-2细胞中的I型IFN信号传导促进Rsad2表达。总之,Rsad2在GC-2细胞中介导BPA诱导的肌动蛋白细胞骨架破坏，为揭示BPA诱导男性生殖毒性的机制提供了数据。
