Role of 5'TG3'-interacting factors (TGIFs) in Vorinostat (HDAC inhibitor)-mediated corneal fibrosis inhibition

摘要: We have previously reported that vorinostat, an FDA-approved, clinically used histone deacetylase (HDAC) inhibitor, attenuates corneal fibrosis in vivo rabbits by blocking transforming growth factor β (TGFβ). The 5'TG3'-interacting factors (TGIFs) are transcriptional repressors of TGFβ1 signaling via the Smad pathway. present study was designed to explore expression TGIFs human fibroblasts and investigate their role mediating antifibrotic effect vorinostat.Human fibroblast cultures were generated from donor corneas. RNA isolation, cDNA preparation, PCR performed detect presence TGIF1 TGIF2 transcripts. exposed vorinostat (2.5 µM) test its on TGIF mRNA protein levels using qPCR immunoblotting. Myofibroblast formation induced with (5 ng/ml) treatment under serum-free conditions. changes parameters quantified measuring marker α-smooth muscle actin (αSMA) qPCR, immunostaining, Smad2/3/4 knockdowns pre-validated RNAi/siRNAs a commercially available transfection reagent.Human showed TGIF2. Vorinostat caused 2.8-3.3-fold increase 1.4-1.8-fold levels. also significant acetylhistone H3 H4. Vorinostat-induced increases accompanied concurrent decrease fibrosis, as indicated αSMA 83±7.7% 97±5%. RNAi-mediated knockdown Smad2, Smad3, Smad4 markedly attenuated TGFβ1-evoked transdifferentiation myofibroblasts. siRNA-mediated neutralized vorinostat-evoked decreases 31%-45% 12%-23%.Human demonstrate transcription factors. These critical, at least partially, cornea.