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RESEARCH PRODUCT

The combination of PDE4 and PDE5 inhibitors reduces YAP expression in IPF

Giancarlo ForteJorge OliverJuliette AlbertBeatriz BallesterJulio CortijoSonia Contreras

subject

A549 cellmedicine.diagnostic_testSildenafilbusiness.industryMesenchymal stem cellImmunofluorescencemedicine.diseaseIn vitrorespiratory tract diseaseschemistry.chemical_compoundchemistryWestern blotFibrosismedicineCancer researchbusinessRoflumilastmedicine.drug

description

Background: Pathological fibrosis is associated with repeated episodes of injury to alveolar epithelial cells of largely. Yes-associated protein (YAP) is prominently expressed in fibrotic lung. Roflumilast has demonstrated to exhibit anti-fibrosis effects, while PDE5 inhibitors (sildenafil) combinated with PDE4 inhibitors reduce fibrotic-effects secondary to TGFs1. Apparently, no studies of PDE4 and PDE5 inhibitors on YAP expression there are. Objectives: To study the effects from adding a sildenafil to Roflumilast N- oxide (RNO) on TGF-β/SMAD3/YAP pathway in IPF in vitro models. Methods: A549 cells were pre-incubated with therapeutical concentrations of RNO (2nM) and/or sildenafil (10nM) and, followed stimulated with TGFβ1. Different marker of EMT, post-transcriptionals factors, pSMAD3 expression and, YAP expression and YAP co-localization were evaluated by RT-PCR, western blot and immunofluorescence. Results: The combination of RNO and sildenafil significantly reduced TGFβ1- induced gene and protein expression of mesenchymal markers collagen type I and alpha actin smooth muscle, prevented the loss of epitelial markers E-cadherin and ZO-1 and inhibited the change of cell phenotype observed in the EMT. Reduction of YAP expression and co-localization was also potentiated by RNO/sildenafil combination. Consequently, the post-transcripcional factors expression activated by YAP were decreased. pSMAD3 expression was decreased with the RNO/sildenafil combination. Conclusion: RNO/sildenafil combination reduces mesenchymal markers and YAP expression avoiding YAP co-localization into nucleus and the post-transcriptional factors expression involved in IPF.

https://doi.org/10.1183/1393003.congress-2017.pa1042