Nintedanib inhibits TGF-β-induced myofibroblast transdifferentiation in human Tenon's fibroblasts.

Abstract

Purpose:

This study aimed to investigate the effect of nintedanib on the conversion of human Tenon's fibroblasts (HTFs) into myofibroblasts and reveal the molecular mechanisms involved.

Methods:

Primary cultured HTFs were incubated with transforming growth factor β1 (TGF-β1) alone or combined with nintedanib, and cell proliferation and migration were measured by cell counting kit-8 (CCK8) and the scratch wound assay, respectively. HTF contractility was evaluated with a 3D collagen contraction assay. The mRNA and protein levels of α smooth muscle actin (α-SMA) and Snail and the phosphorylation levels of Smad2/3, p38 mitogen-activated protein kinase (p38MAPK), and extracellular signal-regulated kinase ½ (ERK1/2) were determined by quantitative reverse transcription polymerase chain reaction (RT-PCR), western blot, and immunofluorescence staining.

Results:

Nintedanib inhibited the proliferation and migration of HTFs in a dose-dependent manner. Furthermore, nintedanib prevented HTF myofibroblast differentiation via downregulation of mRNA and protein expression of α-SMA and Snail. A three-dimensional (3D) collagen gel contraction assay demonstrated that nintedanib effectively inhibits myofibroblast contraction induced by TGF-β1. Mechanistically, we revealed that nintedanib reduces the TGF-β1-induced phosphorylation of Smad2/3, p38MAPK, and ERK1/2, suggesting that nintedanib acts through both classic and nonclassic signaling pathways of TGF-β1 to prevent HTF activation.

Conclusions:

Our study provides new evidence that nintedanib has potent antifibrotic effects in HTFs and suggests that it may be used as a potential therapeutic agent for subconjunctival fibrosis.

• Authors: Xianchai Lin, Jiamin Wen, Rongjiao Liu, Wuyou Gao, Bo Qu, Minbin Yu
• Journal: Molecular vision (Mol Vis) Vol. 24 Pg. 789-800 ( 2018) ISSN: 1090-0535 [Electronic] United States
• PMID: 30636861 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• ACTA2 protein, human
• Actins
• Indoles
• SMAD2 protein, human
• SMAD3 protein, human
• SNAI1 protein, human
• Smad2 Protein
• Smad3 Protein
• Snail Family Transcription Factors
• TGFB1 protein, human
• Transforming Growth Factor beta1
• MAPK1 protein, human
• Mitogen-Activated Protein Kinase 1
• Mitogen-Activated Protein Kinase 3
• p38 Mitogen-Activated Protein Kinases
• nintedanib

Topics

• Actins (antagonists & inhibitors, genetics, metabolism)
• Cell Movement (drug effects)
• Cell Proliferation (drug effects)
• Cell Transdifferentiation (drug effects)
• Dose-Response Relationship, Drug
• Fibroblasts (cytology, drug effects, metabolism)
• Gene Expression Regulation (drug effects)
• Humans
• Indoles (pharmacology)
• Mitogen-Activated Protein Kinase 1 (genetics, metabolism)
• Mitogen-Activated Protein Kinase 3 (genetics, metabolism)
• Myofibroblasts (cytology, drug effects, metabolism)
• Phosphorylation (drug effects)
• Primary Cell Culture
• Signal Transduction (drug effects, genetics)
• Smad2 Protein (genetics, metabolism)
• Smad3 Protein (genetics, metabolism)
• Snail Family Transcription Factors (antagonists & inhibitors, genetics, metabolism)
• Tenon Capsule (cytology, metabolism)
• Transforming Growth Factor beta1 (antagonists & inhibitors, pharmacology)
• p38 Mitogen-Activated Protein Kinases (genetics, metabolism)