Stimulation of Transforming Growth Factor-β1-Induced Endothelial-To-Mesenchymal Transition and Tissue Fibrosis by Endothelin-1 (ET-1): A Novel Profibrotic Effect of ET-1

TGF-beta-induced endothelial-to-mesenchymal transition (EndoMT) is a newly recognized source of profibrotic activated myofibroblasts and has been suggested to play a role in the pathogenesis of various fibrotic processes. Endothelin-1 (ET-1) has been implicated in the development of tissue fibrosis but its participation in TGF-beta-induced EndoMT has not been studied. Here we evaluated the role of ET-1 on TGF-beta 1-induced EndoMT in immunopurified CD31(+)/CD102(+) murine lung microvascular endothelial cells. The expression levels of alpha-smooth muscle actin (alpha-SMA), of relevant profibrotic genes, and of various transcription factors involved in the EndoMT process were assessed employing quantitative RT-PCR, immunofluorescence histology and Western blot analysis. TGF-beta 1 caused potent induction of EndoMT whereas ET-1 alone had a minimal effect. However, ET-1 potentiated TGF-beta 1-induced EndoMT and TGF-beta 1-stimulated expression of mesenchymal cell specific and profibrotic genes and proteins. ET-1 also induced expression of the TGF-beta receptor 1 and 2 genes, suggesting a plausible autocrine mechanism to potentiate TGF-beta-mediated EndoMT and fibrosis. Stimulation of TGF-beta 1-induced skin and lung fibrosis by ET-1 was confirmed in vivo in an animal model of TGF-beta 1-induced tissue fibrosis. These results suggest a novel role for ET-1 in the establishment and progression of tissue fibrosis.