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目的研究凝血酶对人肺上皮细胞及正常人肺成纤维细胞释放单核细胞趋化蛋白-1(MCP-1)的调节作用。方法人肺支气管上皮细胞(normal human bronchial epithelial cells,NHBE)、人肺癌上皮细胞系A549及正常人肺成纤维细胞(normal human lung fibroblasts,NH-LF)分别培养于96孔或6孔板培养板内,凝血酶诱导4h和24h后,收集上清液及细胞裂解液。ELISA方法检测MCP-1蛋白水平;实时荧光定量RT-PCR技术检测MCP-1mRNA水平。结果凝血酶可诱导NHBE、A549及NH-LF细胞释放MCP-1,但NH-LF细胞的MCP-1释放量远远高于NHBE及A549细胞;凝血酶还可诱导NH-LF细胞mRNA表达。结论人肺成纤维细胞可能是肺内表达MCP-1的主要来源细胞,因此在肺纤维化时可能起重要作用。
Objective To study the regulatory effect of thrombin on the release of monocyte chemoattractant protein-1 (MCP-1) from human lung epithelial cells and normal human lung fibroblasts. Methods Normal human bronchial epithelial cells (NHBE), human lung cancer epithelial cell line A549 and normal human lung fibroblasts (NH-LF) were cultured in 96-well or 6-well plates Within 4 h and 24 h after thrombin induction, supernatants and cell lysates were collected. The level of MCP-1 mRNA was detected by ELISA and the level of MCP-1 mRNA by real-time fluorescence quantitative RT-PCR. Results Thrombin induced the release of MCP-1 from NHBE, A549 and NH-LF cells, but the release of MCP-1 from NH-LF cells was much higher than that from NHBE and A549 cells. Thrombin also induced the mRNA expression of NH-LF cells. Conclusion Human lung fibroblasts may be the main source cells for the expression of MCP-1 in the lung and may play an important role in pulmonary fibrosis.