雌激素通过抑制 Smad 反应元件来减弱 TGF-β1 诱导的大鼠尿道平滑肌细胞的弹性生成。
Estrogen attenuates TGF-β1 induced elastogenesis in rat urethral smooth muscle cells by inhibiting Smad response elements.
机构信息
Knuppe Molecular Urology Laboratory, Department of Urology, School of Medicine, University of California-San Francisco, San Francisco, California.
Knuppe Molecular Urology Laboratory, Department of Urology, School of Medicine, University of California-San Francisco, San Francisco, California; Department of Urology, Peking University First Hospital and Institute of Urology, Peking University, Beijing, People's Republic of China.
出版信息
J Urol. 2015 Jun;193(6):2131-7. doi: 10.1016/j.juro.2014.12.085. Epub 2014 Dec 20.
PURPOSE
We investigated the effect and mechanism of estrogen on elastogenesis in urethral smooth muscle cells in vitro.
MATERIALS AND METHODS
Urethral smooth muscle cells were isolated from normal adult female rats. For elastogenesis assay cells were treated with TGF-β1, the potent TGF-β1 receptor inhibitor SB431542 and estrogen for 2 weeks. Real-time polymerase chain reaction was performed to assay gene expression during this process. Activity of the TGF-β1 responsive elements CAGA(12)-Luc and GCCG(12)-Luc were also assayed. Estrogen receptor and Smad2/3 interaction was evaluated by immunoprecipitation and Western blot.
RESULTS
TGF-β1 induced elastogenesis in rat urethral smooth muscle cells. This effect was partially blocked by estrogen and completely abrogated by SB431542. SB431542 completely inhibited activation of the Smad2/3 response element CAGA(12)-Luc and estrogen significantly inhibited activation. The Smad1/4 response element GCCG(12)-Luc was not affected by SB431542 treatment but estrogen partially inhibited the activation of GCCG(12)-Luc induced by TGF-β1. Estrogen receptor bound to Smad 2 and 3 in vitro.
CONCLUSIONS
Estrogen attenuated TGF-β1 induced elastogenesis via binding of its activated receptor to Smad2/3 to inhibit the TGF-β1 response element in rat urethral smooth muscle cells.
目的
我们研究了雌激素对体外尿道平滑肌细胞弹性生成的作用及其机制。
材料与方法
从正常成年雌性大鼠中分离尿道平滑肌细胞。进行弹性生成测定时，用 TGF-β1、有效的 TGF-β1 受体抑制剂 SB431542 和雌激素处理细胞 2 周。在此过程中进行实时聚合酶链反应以检测基因表达。还测定了 TGF-β1 反应元件 CAGA(12)-Luc 和 GCCG(12)-Luc 的活性。通过免疫沉淀和 Western blot 评估雌激素受体和 Smad2/3 相互作用。
结果
TGF-β1 诱导大鼠尿道平滑肌细胞弹性生成。这种作用部分被雌激素阻断，完全被 SB431542 阻断。SB431542 完全抑制 Smad2/3 反应元件 CAGA(12)-Luc 的激活，雌激素显著抑制其激活。Smad1/4 反应元件 GCCG(12)-Luc 不受 SB431542 处理的影响，但雌激素部分抑制 TGF-β1 诱导的 GCCG(12)-Luc 激活。雌激素受体在体外与 Smad 2 和 3 结合。
结论
雌激素通过其激活的受体与 Smad2/3 结合，抑制大鼠尿道平滑肌细胞中 TGF-β1 反应元件，从而减弱 TGF-β1 诱导的弹性生成。
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