Chronic microaspiration of bile acids induces lung fibrosis through multiple mechanisms in rats

Gastroesophageal reflux (GER) and microaspiration of duodenogastric refluxate have been recognized as a risk factor for pulmonary fibrosis. Recent evidence suggests that bile acid microaspiration may contribute to the development of lung fibrosis. However, the molecular evidence is scarce and the underlying mechanisms remain to be elucidated. We have recently demonstrated that bile acids induce activation of alveolar epithelial cells (AECs) and lung fibroblasts in vitro . In the present study, a rat model of bile acid microaspiration was established by weekly intratracheal instillation of three major bile acids including chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), and lithocholic acid (LCA). Repeated microaspiration of CDCA, DCA, and LCA caused fibrotic changes, including alveolar wall thickening and extensive collagen deposition, in rat lungs. Bile acid microaspiration also induced alveolar epithelial–mesenchymal transition (EMT), as indicated by up-regulation of mesenchymal markers α-smooth muscle actin (α-SMA) and vimentin, as well as down-regulaton of epithelial markers E-cadherin and cytokeratin in alveolar epithelium of rat lungs. The expression of fibrogenic mediators, including transforming growth factor-β1 (TGF-β1), connective tissue growth factor (CTGF), basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), and periostin, was significantly elevated in rat lungs exposed to microaspiration of bile acids. Furthermore, microaspiration of bile acids also induced p-Smad3 and farnesoid X receptor (FXR) expression in rat lungs. Our findings suggest that microaspiration of bile acids could promote the development of pulmonary fibrosis in vivo , possibly via stimulating fibrogenic mediator expression and activating TGF-β1/Smad3 signaling and FXR. * α-SMA, : α-smooth muscle actin; AEC, : alveolar epithelial cell; BALF, : bronchoalveolar lavage fluid; bFGF, : basic fibroblast growth factor; BOS, : bronchiolitis obliterans syndrome; CDCA, : chenodeoxycholic acid; CTGF, : connective tissue growth factor; DCA, : deoxycholic acid; ECM, : excessive extracellular matrix; EMT, : epithelial–mesenchymal transition; FXR, : farnesoid X receptor; GER, : gastroesophageal reflux; H&E, : hematoxylin and eosin; HRP, : horseradish peroxidase; IHC, : immunohistochemical; IOD, : integrated optical density; IPF, : idiopathic pulmonary fibrosis; LCA, : lithocholic acid; OB, : obliterative bronchiolitis; TGF-β1, : transforming growth factor-β1; VEGF, : vascular endothelial growth factor