KERATINOCYTE GROWTH-FACTOR AND HEPATOCYTE GROWTH-FACTOR SCATTER FACTOR ARE HEPARIN-BINDING GROWTH-FACTORS FOR ALVEOLAR TYPE-II CELLS IN FIBROBLAST-CONDITIONED MEDIUM
KERATINOCYTE GROWTH-FACTOR AND HEPATOCYTE GROWTH-FACTOR SCATTER FACTOR ARE HEPARIN-BINDING GROWTH-FACTORS FOR ALVEOLAR TYPE-II CELLS IN FIBROBLAST-CONDITIONED MEDIUM
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DOI：
10.1172/jci116673
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发表时间：
1993-08-01
影响因子：
15.9
通讯作者：
MASON, RJ
中科院分区：
文献类型：
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作者：
PANOS, RJ;RUBIN, JS;MASON, RJ
上皮-间充质相互作用介导正常肺生长和发育的各个方面，并且在肺损伤后正常肺泡结构的恢复中是重要的。为了确定成纤维细胞是否是肺泡II型细胞可溶性生长因子的来源，我们研究了成纤维细胞条件培养基（CM）对肺泡II型细胞DNA合成的影响。通过冻干将来自融合的成人肺成纤维细胞的无血清CM浓缩五倍。通过弹性蛋白酶解离从成年大鼠中分离II型细胞，并在培养的第1天至第3天用[H-3]胸苷和不同稀释度的浓缩CM和血清孵育。刺激II型细胞的DNA合成成纤维细胞-CM是最大的条件后48小时，需要血清的存在。CM的活性通过煮沸和用胰蛋白酶、胃蛋白酶或二硫苏糖醇处理而消除，并且与饱和浓度的酸性成纤维细胞生长因子、表皮生长因子和胰岛素一起添加。生长因子活性与肝素-琼脂糖结合，并用0.6和1.0 M NaCl洗脱。中和抗体的研究表明，在0.6和1.0 M NaCl馏分中分离的主要有丝分裂原角质形成细胞生长因子（KGF，成纤维细胞生长因子7）和肝细胞生长因子/分散因子（HGF/SF），分别。HGF/SF被证明在粗CM和KGF中检测到0.6 M NaCl洗脱液通过免疫印迹。北方印迹分析证实肺成纤维细胞表达KGF和HGF/SF转录本。人重组KGF和HGF/SF诱导大鼠肺泡II型细胞DNA合成的浓度和血清依赖性增加。我们得出结论，成人肺成纤维细胞产生至少两种可溶性肝素结合生长因子，KGF和HGF/SF，促进大鼠肺泡II型细胞在原代培养的DNA合成和增殖。KGF和HGF/SF可能是肺生长过程中和肺损伤后肺泡II型细胞增殖的重要刺激因素。
Epithelial-mesenchymal interactions mediate aspects of normal lung growth and development and are important in the restoration of normal alveolar architecture after lung injury. To determine if fibroblasts are a source of soluble growth factors for alveolar type II cells, we investigated the effect of fibroblast-conditioned medium (CM) on alveolar type II cell DNA synthesis. Serum-free CM from confluent adult human lung fibroblasts was concentrated fivefold by lyophilization. Type II cells were isolated from adult rats by elastase dissociation and incubated with [H-3]thymidine and varying dilutions of concentrated CM and serum from day 1 to 3 of culture. Stimulation of type II cell DNA synthesis by fibroblast-CM was maximal after 48 h of conditioning and required the presence of serum. The activity of the CM was eliminated by boiling and by treatment with trypsin, pepsin, or dithiothreitol and was additive with saturating concentrations of acidic fibroblast growth factor, epidermal growth factor, and insulin. The growth factor activity bound to heparin-Sepharose and was eluted with 0.6 and 1.0 M NaCl. Neutralizing antibody studies demonstrated that the primary mitogens isolated in the 0.6 and 1.0 M NaCl fractions were keratinocyte growth factor (KGF, fibroblast growth factor 7) and hepatocyte growth factor/scatter factor (HGF/SF), respectively. HGF/SF was demonstrated in the crude CM and KGF was detected in the 0.6 M NaCl eluent by immunoblotting. Northern blot analysis confirmed that the lung fibroblasts expressed both KGF and HGF/SF transcripts. Human recombinant KGF and HGF/SF induced a concentration- and serum-dependent increase in rat alveolar type II cell DNA synthesis. We conclude that adult human lung fibroblasts produce at least two soluble heparin-binding growth factors, KGF and HGF/SF, which promote DNA synthesis and proliferation of rat alveolar type II cells in primary culture. KGF and HGF/SF may be important stimuli for alveolar type II cell proliferation during lung growth and after lung injury.