Overexpression of ALK5 Induces Human Tenon’s Capsule Fibroblasts Transdifferentiation and Fibrosis In Vitro

ABSTRACTPurpose: To investigate the involvement of activin receptor-like kinase 5 (ALK5) in human Tenon’s capsule fibroblasts (HTFs) transdifferentiation and fibrosis.Methods: (1) Cultured HTFs were treated with transforming growth factor beta 1 (TGF-β1) at different concentrations for different durations, mRNA expression of ALK5 and plasminogen activator inhibitor-1 (PAI-1) was measured by quantitative polymerase chain reaction (PCR) while protein expression of ALK5, α-smooth muscle actin (α-SMA), and extracellular matrix deposition including fibronectin (FN) and collagen I (Col1) was assessed by western blot. HTFs with or without TGF-β1 were also treated with an ALK5 activity inhibitor, SB-431542, and fibrosis-related genes were assessed.(2) HTFs were transduced with ALK5 lentivirus (ALK5-OE group) or empty lentivirus (NC-OE) with or without the treatment of SB-431542. Protein expression of ALK5, α-SMA, FN, and Col1 was evaluated.(3) HTFs in the ALK5-OE group and NC-OE group were subjected to a scratch-...