Inhibition of Signaling Impedes Conversion of Human Tenon's Fibroblasts into Myofibroblasts Via Suppressing Signaling.

Lysophosphatidic acid (LPA) is a growth factor-like phospholipid that has been recognized as a profibrotic mediator in numerous tissues, yet, whether it plays a role in subconjunctival fibrosis remains to be investigated. Therefore, this study was designed to examine the effect of signaling inhibitor, Ki16425 on the conversion of human Tenon's fibroblasts (HTFs) into myofibroblasts. Primary cultured HTFs were incubated with transforming growth factor-β1 () alone or combined with Ki16425, the cell proliferation and migration were measured by Cell Counting Kit-8 and the scratch wound assay, respectively. HTFs contractility was evaluated with 3-dimensional (3D) Collagen Contraction assay. The mRNA and protein levels of -smooth muscle actin (), Snail and the phosphorylation levels of , , and were determined by real-time quantitative polymerase chain reaction (RT-qPCR), western blot, and immunofluorescence staining. Ki16425 significantly prevent the proliferation and migration of Tenon's fibroblasts (HTFs) in a dose-dependent manner. Furthermore, Ki16425 blocked HTFs myofibroblast differentiation via downregulation of mRNA and protein expression of . 3D collagen gel contraction assay demonstrated that Ki16425 effectively inhibits myofibroblast contraction induced by . Mechanistically, we revealed that Ki16425 reduces but not or phosphorylation by . By using an -specific inhibitor, AM095, we confirmed that signaling but not or is involved in induced HTFs activation. Our results show that inhibition of signaling presents potent antifibrotic effect in HTFs, which may serve as a promising intervention strategy for preventing subconjunctival fibrosis caused by glaucoma filtration surgery.