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Involvement of Rho-associated coiled-coil kinase signaling inhibition in TGF-β1/Smad2, 3 signal transduction in vitro

Zhao-Hui Feng1, Xiao-Hui Zhang1, Jia-Qi Zhao1

  • 1Department of Ophthalmology, the Second Affiliated Hospital of Xi'an Jiaotong University Medical College, Xi'an 710004, Shaanxi Province, China.

International Journal of Ophthalmology
|December 21, 2017
PubMed
Abstract

Insights

Y-27632, a ROCK inhibitor, effectively reduced ocular fibroblast proliferation and suppressed TGF-β1 signaling. This suggests ROCK inhibition is a promising therapeutic strategy for filtration channel fibrosis.

Area of Science:

  • Ophthalmology
  • Cell Biology
  • Biochemistry

Background:

  • Fibrosis of the ocular filtration channel is a significant cause of glaucoma treatment failure.
  • Transforming growth factor-beta1 (TGF-β1) signaling, involving Smad2 and Smad3, plays a critical role in fibroblast activation and extracellular matrix deposition.
  • Rho-associated coiled-coil kinase (ROCK) is implicated in cellular processes contributing to fibrosis.

Purpose of the Study:

  • To investigate the inhibitory effects of Y-27632, a selective ROCK inhibitor, on TGF-β1/Smad2,3 signal transduction pathways in ocular Tenon's capsule fibroblasts (OTFs).
  • To evaluate the potential of targeting ROCK signaling as a therapeutic approach for filtration channel fibrosis.

Main Methods:

  • Primary OTFs were cultured in vitro.
  • Cell proliferation was assessed using MTT assays.
  • Gene and protein expression levels of Smad2, Smad3, and phosphorylated Smad2/3 were analyzed by RT-PCR and Western blotting.
  • Alpha-smooth muscle actin (α-SMA) expression was quantified following siRNA transfection and Y-27632 treatment.

Main Results:

  • Y-27632 significantly inhibited lysophosphatidic acid (LPA)-stimulated OTF proliferation.
  • Y-27632 suppressed TGF-β1-induced Smad2 mRNA expression and Smad2/3 protein expression and phosphorylation.
  • While siRNA targeting Smad2/3 reduced α-SMA expression, Y-27632 demonstrated a more potent inhibitory effect.

Conclusions:

  • ROCK signaling inhibition by Y-27632 effectively reduces ocular fibroblast proliferation and key fibrotic signaling pathways.
  • Targeting ROCK may represent a novel therapeutic strategy for preventing or treating filtration channel fibrosis.
Keywords:
ROCKSmadY-27632human ocular Tenon's capsule fibroblaststransforming growth factor-beta1

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