布地奈德在TGF-β1诱导肺成纤维细胞生成VEGF中的调节作用

Effect of Budesonide on transform growth factor-β1 induced vascular endothelial growth factor release by lung fibroblasts

目的:研究糖皮质激素布地奈德对转化生长因子-β1(Transform growth factor-β1,TGF-β1)诱导人胎肺成纤维细胞(Human fetal lung fibroblasts-1,HFL-1)分泌血管内皮生长因子(Vascular endothelial growth factor,VEGF)的影响及作用机制。方法:体外培养HFL,以不同浓度布地奈德预处理,再予以TGF-β1刺激,实验结束时收集细胞上清液经酶联免疫法(Enzyme-linked immunosorbent,ELISA)检测VEGF含量,收集细胞裂解液经Western blot法分别测定磷酸化ERK MAPK(p-ERKMAPK)、磷酸化Smad3(p-Smad3)蛋白表达。结果:TGF-β1能够刺激HFL-1分泌VEGF,且具有时间和浓度依赖效应(P<0.05)。高浓度布地奈德对TGF-β1诱导的VEGF分泌具有抑制作用(P<0.05),同时,布地奈德预处理后p-Smad3及p-ERK MAPK蛋白表达亦明显下调(P<0.05)。结论:布地奈德可能通过阻止Smad3/ERK MAPK信号级联磷酸化从而抑制TGF-β1诱导的VEGF分泌,揭示了糖皮质激素在气道炎症疾病中新的作用机制。

Objective：To observe the effect of glucocorticoid（Budesonide） on TGF-β1 induced human fetal lung fibroblasts-1 （HFL-1 ） secretion and vascular endothelial growth factor（VEGF） pro- duction and to discuss its mechanism. Methods：HFL-1 were cultured in vitro and were treated with various concentrations of Budesonide in the presence of TGF-β1 stimulation. Post-culture media were collected and VEGF content was tested by enzyme- linked immunosorbent（ELISA）. The cell lysates were subjected to Western blot analysis to test the protein expressions of phosphor- ERK MAPK and phosphor-Smad3. Results：Budesonide can induce VEGF release by HFL-1 in a dose-dependent manner（P〈0.05）. Budesonide with high concentration can suppress the TGF-β1 induced VEGF release（P〈0.05）. The protein expressions of phosphor- ERK MAPK and phosphor-Smad3 were decreased after Budesonide pre-treatment（P〈0.05）. Conclusions：Budesonide may reduce TGF-β1 induced VEGF production in the lung,probably through the Smad/ERK signaling pathway,which may provide new sight into the molecular mechanism underlying glucocorticoids therapy for airway inflammatory diseases.