Anti-fibrotic effects of pirfenidone and rapamycin in primary IPF fibroblasts and human alveolar epithelial cells

Abstract

BackgroundPirfenidone, a pleiotropic anti-fibrotic treatment, has been shown to slow down disease progression of idiopathic pulmonary fibrosis (IPF), a fatal and devastating lung disease. Rapamycin, an inhibitor of fibroblast proliferation could be a potential anti-fibrotic drug to improve the effects of pirfenidone.MethodsPrimary lung fibroblasts from IPF patients and human alveolar epithelial cells (A549) were treated in vitro with pirfenidone and rapamycin in the presence or absence of transforming growth factor β1 (TGF−β). Extracellular matrix protein and gene expression of markers involved in lung fibrosis (tenascin-c, fibronectin, collagen I [COL1A1], collagen III [COL3A1] and α-smooth muscle actin [α-SMA]) were analyzed. A cell migration assay in pirfenidone, rapamycin and TGF−β-containing media was performed.ResultsGene and protein expression of tenascin-c and fibronectin of fibrotic fibroblasts were reduced by pirfenidone or rapamycin treatment. Pirfenidone-rapamycin treatment did not revert the epithelial to mesenchymal transition pathway activated by TGF−β. However, the drug combination significantly abrogated fibroblast to myofibroblast transition. The inhibitory effect of pirfenidone on fibroblast migration in the scratch-wound assay was potentiated by rapamycin combination.ConclusionsThese findings indicate that the combination of pirfenidone and rapamycin widen the inhibition range of fibrogenic markers and prevents fibroblast migration. These results would open a new line of research for an anti-fibrotic combination therapeutic approach.

Analysis of the Interaction Network of Hub miRNAs-Hub Genes, Being Involved in Idiopathic Pulmonary Fibers and Its Emerging Role in Non-small Cell Lung Cancer

“…were considered to be significantly related to IPF and NSCLC. Focal adhesion was considered as a key pathway shared by blue module and turquoise module, and many gens/proteins have been considered to be involved in the progression of IPF through disordering focal adhesion (Gimenez et al, 2017;Kathiriya et al, 2017;Molina-Molina et al, 2018). For example, it has been reported that decreased expression of collagen VI, an important kind of protein of ECM, would upregulate the focal adhesion (Knueppel et al, 2018).…”

“…Previous studies have revealed that ECM protein expression plays an important role in the fibrotic process in IPF lungs (Vicens-Zygmunt et al, 2015). Excessive accumulation of ECM in the alveolar parenchyma and progressive scarring of lung tissue are major characteristics of IPF (Knudsen et al, 2017), and some studies have used this protein expression level as a criterion for evaluating treatment outcomes (Molina-Molina et al, 2018;Mullenbrock et al, 2018). Altogether, migration is strongly influenced by topology and composition of the ECM including integrin ligands, and the hub gens and hub miRNAs might play an important role in IPF progression with the change of ECM.…”

Sirolimus (rapamycin) for the targeted treatment of the fibrotic sequelae of Graves’ orbitopathy

“…Sirolimus acts by binding to FK-binding-protein-12 to inhibit a subunit protein of the mechanistic target of rapamycin called complex 1 (mTORC1), which is essential for T cell activation and which has also been implicated in adipogenesis [1]. In vitro studies have additionally demonstrated anti-fibroblast effects of rapamycin, including reduction in fibroblast migration and reduced transition to myofibroblasts [2], proliferation, collagen expression, interleukin-16 (IL-16) production [3] and cellular metabolism [4]. In vivo, murine experiments indicate reduced wound repair and fibrosis with immune suppression [5] as well as reduced fibrosis in a variety of organ systems including kidney [6] and heart [7] with rapamycin treatment.…”

“…Overall, the evidence suggests that mTOR plays a significant role in the control of abnormal fibroblast proliferation, differentiation and extrusion of stromal contents including collagen [2,4]. Because of its attenuation of adipogenesis and effect on fibroblast proliferation and fibrosis-processes all implicated in the pathogenesis of Graves' orbitopathy (GO) [1]-we sought to determine whether rapamycin could be a potential adjunct in the treatment of thyroid eye disease (TED).…”

Prevention of tracheal stenosis with pirfenidone after tracheotomy: An experimental study

“…Although its exact mechanism of action has not been established yet, it is suggested that it acts by decreasing the production rate of TGFβ-1 (the growth factor beta), TNFα (tumor necrosis factor alpha), and IL-1β (Interleukin-1 beta) in the in vitro and in vivo fibrotic disease models. [4][5][6] Despite the use of pirfenidone in primarily pulmonary fibrosis, as well as the experimental studies using it in the disorders manifesting with fibrosis-including the cutaneous scars, 4 vocal fold scars, 7 abdominal adhesions, 8 intestinal 9 and esophageal strictures, 10 and benign biliary strictures 11 -the number of studies on tracheal stenosis is limited. 3,12,13 This study aimed to evaluate the effect of pirfenidone on the development of tracheal stenosis in the rats intubated with a tracheal tube.…”