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

摘要:

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-ERK MAPK）、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。

Abstract:

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.