Inhibition of Apoptosis Signal-Regulating Kinase 1 Attenuates Myocyte Hypertrophy and Fibroblast Collagen Synthesis

Heart Lung Circ. 2019 Mar;28(3):495-504. doi: 10.1016/j.hlc.2017.12.001. Epub 2017 Dec 14.

Abstract

Background: Cardiac remodelling is a dynamic process whereby structural and functional changes occur within the heart in response to injury or inflammation. Recent studies have demonstrated reactive oxygen species sensitive MAPK, apoptosis signal-regulating kinase 1 (ASK1) plays a critical role in cardiac remodelling. This study aims to determine the effectiveness of small molecule ASK1 inhibitors on these processes and their therapeutic potential.

Methods: Neonatal rat cardiac fibroblasts (NCF) were pre-treated with ASK1 inhibitors, G2261818A (G226) and G2358939A (G235), for 2hours before stimulated with 100nM angiotensin II (AngII), 10μM indoxyl sulphate (IS) or 10ng/ml transforming growth factor β1 (TGFβ1) for 48hours. Neonatal rat cardiac myocytes (NCM) were pre-treated with G226 and G235 for 2hours before being stimulated with 100nM AngII for 60hours, 10μM IS, 10ng/ml interleukin 1β (IL-1β) or tumour necrosis factor α (TNFα) for 48hours. 3H-proline and 3H-leucine incorporation was used to assess collagen turnover and hypertrophy, respectively. Pro-fibrotic, pro-hypertrophic and THP-1 inflammatory cytokine gene expressions were determined by RT-PCR.

Results: Both G226 and G235 dose-dependently attenuated AngII-, IS-, IL-1β- and TNFα-stimulated NCM hypertrophy and hypertrophic gene expression, IS-, AngII- and TGFβ1-stimulated NCF collagen synthesis and AngII- and TGFβ1-stimulated pro-fibrotic gene expression. Inhibition of ASK1 by G226 and G235 inhibited lipopolysaccharides-stimulated inflammatory cytokine gene expression in THP-1 cells.

Conclusions: Selective ASK1 inhibition confers anti-hypertrophic and anti-fibrotic effects in cardiac cells, and anti-inflammation in monocytic cells. ASK1 inhibitors may represent novel therapeutic agents to alleviate cardiac remodelling post cardiac injury where hypertrophy, fibrosis and inflammation play critical roles.

Keywords: ASK1 inhibitors; Apoptosis signal-regulating kinase 1; Cardiac hypertrophy; Cardiac remodelling; Collagen synthesis.

MeSH terms

  • Animals
  • Animals, Newborn
  • Cardiomegaly / genetics*
  • Cardiomegaly / metabolism
  • Cardiomegaly / pathology
  • Cells, Cultured
  • Collagen / biosynthesis*
  • Disease Models, Animal
  • Fibroblasts / metabolism
  • Gene Expression Regulation, Developmental*
  • MAP Kinase Kinase Kinase 5 / antagonists & inhibitors
  • MAP Kinase Kinase Kinase 5 / biosynthesis
  • MAP Kinase Kinase Kinase 5 / genetics*
  • Myocytes, Cardiac / metabolism
  • Myocytes, Cardiac / pathology*
  • Polymerase Chain Reaction
  • RNA / genetics*
  • Rats
  • Rats, Sprague-Dawley
  • Signal Transduction

Substances

  • RNA
  • Collagen
  • MAP Kinase Kinase Kinase 5