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为探讨肺内神经肽在气道损伤修复中的作用 ,采用blind wellBoydenchamber测定原代培养的支气管上皮细胞 (bronchialepithelialcells,BEC)趋化性 ,观察血管活性肠肽 (vasoactiveintestinalpeptide ,VIP)对BEC趋化迁移的影响及其机制 ,并测定经热应激后BEC分泌VIP及表达VIP受体 (vasoactiveintestinalpeptidereceptor,VIPR)的变化。结果显示 :(1)以胰岛素作为趋化因子所建立的BEC趋化性测定方法稳定 ,重现性好 (r =0 970 3,P <0 0 1) ;(2 )VIP (0 0 0 1～ 1μmol/L)均显示剂量依赖性地增强BEC的趋化迁移 ,其效应可被钙调蛋白阻断剂及蛋白激酶C阻断剂有效地抑制 (P <0 0 1) ;(3) 4 2℃、30min热应激后BEC分泌VIP (P <0 0 1)及表达VIPR明显增加 (P <0 0 5 )。实验表明 :肺内神经肽VIP可增强BEC的趋化迁移 ,其细胞内信号转导途径与钙调蛋白及蛋白激酶C有关。而热应激时VIP及VIPR的高表达进一步提示局部微环境的VIP可能是气道上皮损伤修复网络中的重要分子
To investigate the role of intranasal neuropeptides in the repair of airway injury, the chemotaxis of primary cultured bronchial epithelial cells (BEC) was measured by blind well-chamber positive volume, and the chemotactic migration of BEC was observed by vasoactive intestinal peptide (VIP) And its mechanism. The changes of VIP secretion and vasoactive intestinal peptide receptor (VIPR) secretion by BEC after heat stress were measured. The results showed that: (1) The chemotactic assay of BEC established by using insulin as a chemokine was stable and reproducible (r = 0 970 3, P 0 01); (2) VIP ~ 1 μmol / L) showed a dose-dependent increase in the chemotactic migration of BEC, the effect of which was inhibited by calmodulin and protein kinase C blockers (P <0.01); (3) 4 After heat stress for 30 minutes at 2 ℃, BEC secreted VIP (P <0.01) and VIPR increased (P <0.05). Experiments show that VIP in the lung can enhance the chemotactic migration of BEC, and its intracellular signal transduction pathway is related to calmodulin and protein kinase C. However, the high expression of VIP and VIPR in heat stress further suggests that VIP in the local microenvironment may be an important molecule in the airway epithelial injury repair network