›› 2009, Vol. 40 ›› Issue (6): 919-922.doi: 10.3969/j.issn.0529-1356.2009.06.014

• 论著 • 上一篇    下一篇

细胞外调节蛋白激酶信号通路参与机械牵张诱导肺泡上皮细胞高迁移率族蛋白B1的表达调控

丁宁1; 肖慧2; 高巨3 ;许立新1*; 佘守章1   

  1. 1.广州市第一人民医院 麻醉科和麻醉学实验室; 2.门诊部,广州 510180; 3.广州中医药大学第二附属医院麻醉科,广州 510120
  • 收稿日期:2008-10-24 修回日期:2008-12-01 出版日期:2009-12-06
  • 通讯作者: 许立新

ERK signal is pathway involved in mechanical stretch induced HMGB1 expression in alveolar epithelial cells

  1. 1. Department of Anesthesiology and Anesthesiology Laboratory; 2. Department of Outpatient, Guangzhou First Municipal People’s Hospital, Guangzhou 510180, China; 3.Department of Anesthesiology, the Second Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, Guangzhou 510120, China)
  • Received:2008-10-24 Revised:2008-12-01 Online:2009-12-06
  • Contact: XU Li-xin

关键词: 机械牵张, 高迁移率族蛋白B1, 细胞外调节蛋白激酶, 免疫组织化学, 反转录-聚合酶链反应, 免疫印迹法, 肺泡上皮细胞

Abstract: Objective To investigate the role of extracellular regulated protein kinase (ERK) signal pathway in mechanical stretch induced high mobility group box 1 protein (HMGB1) expression on alveolar epithelial cells (A549). MethodsA549 cells were cultured and seeded at 1×10SUP>5 /SUP>cells/ml in 6-well Bioflex cell culture plates. Subsequently, the cells were exposed to cyclic mechanical stretch at 14% (group B) elongation for 4 hours using Flexercell 4000T cell stretching unit. In group C, cells were pretreated with PD98059 for 2 hours before mechanical stretch. Cells in group A without stretch were served as control. The expression of HMGB1 protein and mRNA in A549 cells were detected by immunocytochemisty staining and RT-PCR, respectively. ERK activity was measured by Western blotting method. Results Immunocytochemisty staining indicated that the expression of HMGB1 protein in A549 cells was increased obviously in group B (EM>P/EM><0.05) and decreased in group C (EM>P/EM><0.05). Polymerase chain reaction (RT-PCR) showed that the expression of HMGB1 mRNA was also significantly increased in group B (EM>P/EM><0.05) and decreased in group C (EM>P/EM><0.05). Western blotting analysis confirmed the activation of ERK in A549 cells by mechanical stretch (EM>P/EM><0.05). PD98059, an inhibitor of ERK, might significantly inhibit mechanical stretch induced HMGB1 protein and mRNA expression in A549 cells (P<0.05). Conclusion Mechanical stretch could regulate the expression of HMGB1 gene and protein in A549 cells through ERK signal pathway.

Key words: Mechanical stretch, High-mobility group box-1, ERK, Alveolar epithelial cell, Immunohistochemistry, RT-PCR, Western blotting

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