Cytokine receptor-like factor 1 (CRLF1) promotes cardiac fibrosis via ERK1/2 signaling pathway

J Zhejiang Univ Sci B. 2023 Aug 15;24(8):682-697. doi: 10.1631/jzus.B2200506.
[Article in English, Chinese]

Abstract

Cardiac fibrosis is a cause of morbidity and mortality in people with heart disease. Anti-fibrosis treatment is a significant therapy for heart disease, but there is still no thorough understanding of fibrotic mechanisms. This study was carried out to ascertain the functions of cytokine receptor-like factor 1 (CRLF1) in cardiac fibrosis and clarify its regulatory mechanisms. We found that CRLF1 was expressed predominantly in cardiac fibroblasts. Its expression was up-regulated not only in a mouse heart fibrotic model induced by myocardial infarction, but also in mouse and human cardiac fibroblasts provoked by transforming growth factor-‍β1 (TGF‍-‍β1). Gain- and loss-of-function experiments of CRLF1 were carried out in neonatal mice cardiac fibroblasts (NMCFs) with or without TGF-‍β1 stimulation. CRLF1 overexpression increased cell viability, collagen production, cell proliferation capacity, and myofibroblast transformation of NMCFs with or without TGF‍-‍β1 stimulation, while silencing of CRLF1 had the opposite effects. An inhibitor of the extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway and different inhibitors of TGF-‍β1 signaling cascades, comprising mothers against decapentaplegic homolog (SMAD)‍-dependent and SMAD-independent pathways, were applied to investigate the mechanisms involved. CRLF1 exerted its functions by activating the ERK1/2 signaling pathway. Furthermore, the SMAD-dependent pathway, not the SMAD-independent pathway, was responsible for CRLF1 up-regulation in NMCFs treated with TGF-‍β1. In summary, activation of the TGF-‍β1/SMAD signaling pathway in cardiac fibrosis increased CRLF1 expression. CRLF1 then aggravated cardiac fibrosis by activating the ERK1/2 signaling pathway. CRLF1 could become a novel potential target for intervention and remedy of cardiac fibrosis.

心脏纤维化是心脏疾病患者发病和死亡的原因之一。抗纤维化治疗是一种治疗心脏疾病的重要手段,但目前对纤维化的机制仍缺乏深入了解。本研究旨在确定细胞因子受体样因子1(CRLF1)在心脏纤维化中的功能并阐明其调节机制。我们发现CRLF1主要在心脏成纤维细胞中表达;无论是在心肌梗死诱导的小鼠心脏纤维化模型还是在转化生长因子-β1(TGF-‍β1)刺激的小鼠和人心脏成纤维细胞中,CRLF1表达均上调。本研究在使用或不使用TGF-β1刺激的新生乳鼠心脏成纤维细胞(NMCFs)中开展了CRLF1的功能获得和丧失实验。在TGF-β1刺激或不刺激的情况下,CRLF1的过表达均可增加NMCFs的细胞活力、胶原生成、细胞增殖能力及肌成纤维细胞转化,而CRLF1沉默则具有相反效果。应用细胞外信号调节激酶1/2(ERK1/2)信号通路抑制剂以及包括SMAD依赖和非依赖信号在内的不同TGF-β1下游信号通路抑制剂来开展机制研究。CRLF1通过激活ERK1/2信号通路发挥其功能。此外,CRLF1在TGF-β1处理的NMCFs中表达上调是由SMAD依赖性通路介导,而不是SMAD非依赖性通路介导。总而言之,心脏纤维化中TGF-β1/SMAD信号通路的激活增加了CRLF1的表达。随后,CRLF1通过激活ERK1/2信号通路加重了心脏纤维化。因此,CRLF1可作为一个干预和治疗心脏纤维化的新的潜在靶点。.

心脏纤维化是心脏疾病患者发病和死亡的原因之一。抗纤维化治疗是一种治疗心脏疾病的重要手段,但目前对纤维化的机制仍缺乏深入了解。本研究旨在确定细胞因子受体样因子1(CRLF1)在心脏纤维化中的功能并阐明其调节机制。我们发现CRLF1主要在心脏成纤维细胞中表达;无论是在心肌梗死诱导的小鼠心脏纤维化模型还是在转化生长因子-β1(TGF-‍β1)刺激的小鼠和人心脏成纤维细胞中,CRLF1表达均上调。本研究在使用或不使用TGF-β1刺激的新生乳鼠心脏成纤维细胞(NMCFs)中开展了CRLF1的功能获得和丧失实验。在TGF-β1刺激或不刺激的情况下,CRLF1的过表达均可增加NMCFs的细胞活力、胶原生成、细胞增殖能力及肌成纤维细胞转化,而CRLF1沉默则具有相反效果。应用细胞外信号调节激酶1/2(ERK1/2)信号通路抑制剂以及包括SMAD依赖和非依赖信号在内的不同TGF-β1下游信号通路抑制剂来开展机制研究。CRLF1通过激活ERK1/2信号通路发挥其功能。此外,CRLF1在TGF-β1处理的NMCFs中表达上调是由SMAD依赖性通路介导,而不是SMAD非依赖性通路介导。总而言之,心脏纤维化中TGF-β1/SMAD信号通路的激活增加了CRLF1的表达。随后,CRLF1通过激活ERK1/2信号通路加重了心脏纤维化。因此,CRLF1可作为一个干预和治疗心脏纤维化的新的潜在靶点。

Keywords: Cardiac fibrosis; Cytokine receptor-like factor 1 (CRLF1); ERK1/2 signaling pathway; Extracellular matrix (ECM); Myofibroblast transformation; TGF‍-‍β1/SMAD signaling pathway.

MeSH terms

  • Animals
  • Disease Models, Animal
  • Fibroblasts / metabolism
  • Fibrosis
  • Humans
  • MAP Kinase Signaling System*
  • Mice
  • Mitogen-Activated Protein Kinase 3 / metabolism
  • Myocardial Infarction* / metabolism
  • Receptors, Cytokine* / metabolism
  • Signal Transduction
  • Transforming Growth Factor beta1 / pharmacology

Substances

  • Mitogen-Activated Protein Kinase 3
  • Receptors, Cytokine
  • Transforming Growth Factor beta1
  • cytokine-like factor-1