探讨肺纤维化时表达单核细胞趋化蛋白-1的主要效应细胞

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。

The Major Cell Type Responsible for Monocyte Chemoattractant Protein-1 Release in Pulmonary Fibrosis

Objective To investigate the effect of thrombin on monocyte chemoattractant protein-1 （MCP-1） release in human lung epithelial cells and normal human lung fibroblasts.Methods Normal human bronchial epithelial cells （NHBE）,A549 and normal human lung fibroblasts （NH-LF） were cultured in a 96-well or 6-well culture plate.Cells were stimulated with thrombin （10 nmol/L） for 4 and 24 h.Culture supernatants of each well from 96-well plates were collected at 4 h time point.MCP-1 protein levels in supernatants were assessed by ELISA.Cells in 6-well plates were collected for mRNA detection by quantitative real time RT-PCR after 4 and 24 h stimulation with thrombin.Results Thrombin can induce NHBE,A549 and NH-LF to relase MCP-1,and the MCP-1 levels in NHLF are much higher than the MCP-1 levels in both NHBE and A549 cells.Thrombin can also induce MCP-1 mRNA expression in NH-LF cells.Conclusion MCP-1 may be mainly produced by fibroblasts in the lung.Fibroblasts may therefore play an important role in the process of pulmonary fibrosis.