贝前列素抑制Ang II刺激下心肌成纤维细胞中TGF β-Smad信号通路
孟国梁，陈云，张伟
基金项目：江苏省高校自然科学基金项目（12KJB310012）；南通市自然科学基金项目（BK2011044,2012087） 作者简介：孟国梁（1981-），男，讲师，主要研究方向：心血管药理学 通信联系人：张伟（1957-），男，教授，博士生导师，主要研究方向：心血管药理学.

（南通大学药学院，南通 226001）
摘要：目的：探讨前列环素类似物贝前列素对Ang II刺激下心肌成纤维细胞中TGF β-Smad信号通路的影响。方法：体外培养乳大鼠心肌成纤维细胞，贝前列素（10 μM，4 h）预处理后，Ang II（100 nM，24 h）刺激心肌成纤维细胞，Western blot测定转化生长因子β（TGF β）、Smad2和cAMP应答原件结合蛋白（CREB）表达，电泳迁移率实验测定Smad蛋白10 与DNA相互作用，免疫共沉淀观察CBP与Smad2、p-CREB的结合。结果：贝前列素能抑制TGF β表达及Smad2的磷酸化，减弱Smad2与DNA的结合力，增加细胞核中ser133-p-CREB表达。贝前列素预处理亦可增加CBP与磷酸化CREB结合，从而减少其与Smad2结合。结论：贝前列素抑制Ang II刺激下心肌成纤维细胞中TGF β-Smad信号通路。
关键词：药理学；贝前列素；心肌成纤维细胞；转录生长因子β；cAMP应答元件结合蛋白

中图分类号：R96
Beraprost inhibits TGF β-Smad signal pathway in Ang II-stimulated cardiac fibroblasts
MENG Guoliang, CHEN Yun, ZHANG Wei 20
(School of Pharmacy, Nantong University, Nantong 22001)
Abstract: Objective: To investigate the effect of beraprost on TGF β-Smad signal pathway in Ang II-stimulated cardiac fibroblasts. Methods: Neonatal rat cardiac fibroblasts were pre-treated with beraprost (10 μM, 4 h) followed by Ang II (100 nM, 24 h) stimulation. Protein levels of TGF β, Smad2 and CREB were analyzed by Western blot. Electrophoretic mobility shift assays (EMSA) 25 was used to detect the Smad-DNA binding activity. The combination of CBP or Smad2 with p-CREB was investigated by co-immunoprecipitaion assays. Results: Beraprost significantly blocked TGF β expression as well as Smad2 phosphorylation and reduced Smad-DNA binding activity. Beraprost also increased nuclear translocation of phosphorylation of cAMP response element binding protein (CREB) at ser133. More CREB but less Smad2 binding to 30 CREB-binding protein (CBP) after beraprost pre-treatment. Conclusion: Beraprost inhibited TGF β-Smad signal pathway in Ang II-stimulated cardiac fibroblasts.
Key words: Pharmacology; Beraprost; Cardiac fibroblast; Transforming growth factor β; cAMP response element binding protein