Smad2/3 Upregulates the Expression of Vimentin and Affects Its Distribution in DBP-Exposed Sertoli Cells.
Smad2/3 Upregulates the Expression of Vimentin and Affects Its Distribution in DBP-Exposed Sertoli Cells.
复制标题
Smad2/3 上调 Vimentin 的表达并影响其在 DBP 暴露的支持细胞中的分布
作者：
Zhang X;Wang X;Liu T;Mo M;Ao L;Liu J;Cao J;Cui Z
睾丸中的支持细胞（SC）为生殖细胞提供物理和营养支持。邻苯二甲酸二丁酯（DBP）破坏了SC中的波形蛋白细胞骨架，导致SC功能障碍。在先前的研究中，我们发现过氧化物酶体增殖物激活受体α（PPAR α）通过影响波形蛋白在DBP暴露的SC中的磷酸化来影响波形蛋白的分布。在本研究中，我们研究了Smad2/3在DBP暴露的SC中调节波形蛋白表达的作用。我们推测Smad2/3可能通过调节波形蛋白的表达来影响波形蛋白的分布，并与PPAR α之间存在相互作用。Real-time PCR和ChIP-qPCR结果显示，Smad2/3抑制剂SB431542可显著抑制DBP诱导的SC波形蛋白表达。SB431542预处理可下调磷酸化和可溶性波形蛋白。结果表明，预处理后的波形蛋白中Smad2/3活性增加，而抑制剂GW6471抑制Smad2/3活性，SB431542也抑制了波形蛋白中PPAR α的活性，但不能挽救DBP诱导的波形蛋白塌陷。我们的研究结果表明，除了促进波形蛋白的磷酸化，DBP还通过激活SC中的Smad2/3来刺激波形蛋白的表达，从而诱导波形蛋白的不规则分布。
Sertoli cells (SCs) in the testes provide physical and nutritional support to germ cells. The vimentin cytoskeleton in SCs is disrupted by dibutyl phthalate (DBP), which leads to SCs dysfunction. In a previous study, we found that peroxisome proliferator-activated receptor alpha (PPARα) influenced the distribution of vimentin by affecting its phosphorylation in DBP-exposed SCs. In the present study, we investigated the role of Smad2/3 in regulating the expression of vimentin in DBP-exposed SCs. We hypothesized that Smad2/3 affects the distribution of vimentin by regulating its expression and that there is cross talk between Smad2/3 and PPARα. The real-time PCR and ChIP-qPCR results showed that SB431542 (an inhibitor of Smad2/3) could significantly attenuate the expression of vimentin induced by DBP in SCs. Phosphorylated and soluble vimentin were both downregulated by SB431542 pretreatment. WY14643 (an agonist of PPARα) pretreatment stimulated, while GW6471 (an antagonist of PPARα) inhibited, the activity of Smad2/3; SB431542 pretreatment also inhibited the activity of PPARα, but it did not rescue the DBP-induced collapse in vimentin. Our results suggest that, in addition to promoting the phosphorylation of vimentin, DBP also stimulates the expression of vimentin by activating Smad2/3 in SCs and thereby induces irregular vimentin distribution.