University of Chicago, Department of Medicine, Section of Pulmonary and Critical Care Medicine, Chicago, IL, USA.
Suez University, Faculty of Science, Zoology Department, Suez, Egypt.
J Pathol. 2018 Sep;246(1):54-66. doi: 10.1002/path.5106. Epub 2018 Jul 5.
Fibroblast growth factor (FGF) signaling has been implicated in the pathogenesis of pulmonary fibrosis. Mice lacking FGF2 have increased mortality and impaired epithelial recovery after bleomycin exposure, supporting a protective or reparative function following lung injury. To determine whether FGF2 overexpression reduces bleomycin-induced injury, we developed an inducible genetic system to express FGF2 in type II pneumocytes. Double-transgenic (DTG) mice with doxycycline-inducible overexpression of human FGF2 (SPC-rtTA;TRE-hFGF2) or single-transgenic controls were administered intratracheal bleomycin and fed doxycycline chow, starting at either day 0 or day 7. In addition, wild-type mice received intratracheal or intravenous recombinant FGF2, starting at the time of bleomycin treatment. Compared to controls, doxycycline-induced DTG mice had decreased pulmonary fibrosis 21 days after bleomycin, as assessed by gene expression and histology. This beneficial effect was seen when FGF2 overexpression was induced at day 0 or day 7 after bleomycin. FGF2 overexpression did not alter epithelial gene expression, bronchoalveolar lavage cellularity or total protein. In vitro studies using primary mouse and human lung fibroblasts showed that FGF2 strongly inhibited baseline and TGFβ1-induced expression of alpha smooth muscle actin (αSMA), collagen, and connective tissue growth factor. While FGF2 did not suppress phosphorylation of Smad2 or Smad-dependent gene expression, FGF2 inhibited TGFβ1-induced stress fiber formation and serum response factor-dependent gene expression. FGF2 inhibition of stress fiber formation and αSMA requires FGF receptor 1 (FGFR1) and downstream MEK/ERK, but not AKT signaling. In summary, overexpression of FGF2 protects against bleomycin-induced pulmonary fibrosis in vivo and reverses TGFβ1-induced collagen and αSMA expression and stress fiber formation in lung fibroblasts in vitro, without affecting either inflammation or epithelial gene expression. Our results suggest that in the lung, FGF2 is antifibrotic in part through decreased collagen expression and fibroblast to myofibroblast differentiation. Copyright © 2018 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
成纤维细胞生长因子(FGF)信号在肺纤维化的发病机制中起作用。缺乏 FGF2 的小鼠在博来霉素暴露后死亡率增加,上皮细胞恢复受损,这支持了肺损伤后的保护或修复功能。为了确定 FGF2 的过表达是否能减轻博来霉素诱导的损伤,我们开发了一种诱导型遗传系统,以在 II 型肺泡细胞中表达 FGF2。用强力霉素诱导性过表达人 FGF2(SPC-rtTA;TRE-hFGF2)的双转基因(DTG)小鼠或单转基因对照小鼠接受气管内博来霉素给药,并在第 0 天或第 7 天开始给予强力霉素饲料。此外,野生型小鼠在接受博来霉素治疗时接受气管内或静脉内重组 FGF2。与对照组相比,在博来霉素后 21 天,通过基因表达和组织学评估,强力霉素诱导的 DTG 小鼠的肺纤维化程度降低。当在博来霉素后第 0 天或第 7 天诱导 FGF2 过表达时,观察到这种有益作用。FGF2 过表达并未改变上皮基因表达、支气管肺泡灌洗液细胞计数或总蛋白。使用原代小鼠和人肺成纤维细胞的体外研究表明,FGF2 强烈抑制基础状态和 TGFβ1 诱导的α平滑肌肌动蛋白(αSMA)、胶原蛋白和结缔组织生长因子的表达。虽然 FGF2 不抑制 Smad2 磷酸化或 Smad 依赖性基因表达,但 FGF2 抑制 TGFβ1 诱导的应激纤维形成和血清反应因子依赖性基因表达。FGF2 抑制应激纤维形成和αSMA 需要 FGFR1(FGFR1)和下游 MEK/ERK,但不需要 AKT 信号传导。总之,FGF2 的过表达可在体内预防博来霉素诱导的肺纤维化,并在体外逆转 TGFβ1 诱导的肺成纤维细胞中胶原蛋白和αSMA 表达和应激纤维形成,而不影响炎症或上皮基因表达。我们的结果表明,在肺部,FGF2 通过减少胶原蛋白表达和纤维母细胞向肌成纤维细胞分化而具有抗纤维化作用。版权所有©2018 英国和爱尔兰病理学学会。由 John Wiley & Sons,Ltd. 出版。