摘要目的 探讨特发性肺纤维化中成纤维细胞对气道祖细胞增殖功能的调控.方法 制备β-肌动蛋白-绿色荧光蛋白(green fluorescent protein,GFP)小鼠肺组织单细胞悬液,利用流式细胞术分选得到气道祖细胞;通过气道祖细胞与肺成纤维细胞共培养观察气道祖细胞的生长情况.TGF-β受体抑制剂SB431542处理成纤维细胞,测定成纤维细胞分泌生长因子1(fibroblast growth factor 1,FGF1)和成纤维细胞生长因子2(fibroblast growth factor2,FGF2)的变化,观察FGF1/2对气道祖细胞生长的作用.结果 流式细胞术分选得到的气道祖细胞和正常人来源的肺成纤维细胞共培养,8d后克隆形成率为(3.5±1.1)％;而气道祖细胞和特发性肺纤维化患者来源的肺成纤维细胞共培养时,克隆明显减少,克隆形成率仅(0.04±0.04)％,两组比较差异有统计学意义(P＜0.05).荧光显微镜下可见SB431542上调肺成纤维细胞分泌的生长因子FGF1/2的表达,FGF1 mRNA的表达量由对照组的0.0002上调到实验组的0.005 5,FGF2 mRNA的表达量由对照组的0.0008上调到实验组的0.001 5,FGF1/2均促进了气道祖细胞的生长.FGF1/2单独培养气道祖细胞,8d后成功出现克隆,形成率为(0.3±0.1)％.结论 在特发性肺纤维化发展过程中,成纤维细胞分泌的生长因子FGF1/2调控气道祖细胞增殖.

abstractsObjective To investigate the effect of fibroblasts on regulating airway stem cell proliferation in idiopathic pulmonary fibrosis.Methods Lung cell suspension was prepared from β-actinGFP mice.Airway stem cells were obtained by fluorescence activated cell sorting and co-cultured with lung fibroblasts.The fibroblasts were treated with TGF-β inhibitor SB43142.The expression of growth factors FGF1/2 and the effect of FGF1/2 on stem cell proliferation were observed.Results The cloning efficiency of airway stem cells,when co-cultured with normal lung fibroblast cells for 8 days,was (3.5 ± 1.1) ％,while the cloning efficiency was reduced to (0.04 ± 0.04) ％ when co-cultured with lung fibroblasts from idiopathic pulmonary fibrosis patients.The difference between the 2 groups was statistically significant(P =0.002 5).TGF-β receptor inhibitor SB431542 increased lung fibroblast growth factors FGF1/2 expression.FGF1 mRNA expression was increased to the experimental group 0.005 5 from 0.000 2 in the control group.FGF2 mRNA expression of the amount raised to the experimental group 0.000 15 from 0.000 8 in the control group.FGF1/2 promoted the growth of airway stem cells.After FGF1/2 was co-cultured with normal lung fibroblast cells for 8 days,the cloning efficiency of airway stem cells was (0.3 ± 0.1) ％.Conclusion During the development of idiopathic pulmonary fibrosis,fibroblast secreted FGF1/2 regulate airway stem cell proliferation.