An integrated approach to identify critical transcription factors in the protection against hydrogen peroxide-induced oxidative stress by Danhong injection

Free Radic Biol Med. 2017 Nov:112:480-493. doi: 10.1016/j.freeradbiomed.2017.07.002. Epub 2017 Aug 16.

Abstract

Oxidative stress plays a vital role in many pathological processes of the cardiovascular diseases. However, the underlying mechanism remains unclear, especially on a transcription factor (TF) level. In this study, a new method, concatenated tandem array of consensus transcription factor response elements (catTFREs), and an Illumina-based RNA-seq technology were integrated to systematically investigate the role of TFs in hydrogen peroxide (H2O2)-induced oxidative stress in cardiomyocytes; the damage was then rescued by Danhong injection (DHI), a Chinese standardized product approved for cardiovascular diseases treatment. The overall gene expression revealed cell apoptosis and DNA repair were vital for cardiomyocytes in resisting oxidative stress. By comprehensively integrating the transcription activity of TFs and their downstream target genes, an important TFs-target network were constructed and 13 TFs were identified as critical TFs in DHI-mediated protection in H2O2-induced oxidative stress. By using the integrated approach, seven TFs of these 13 TFs were also identified in melatonin-mediated protection in H2O2-induced damage. Furthermore, the transcription activity of DNA-(apurinic or apyrimidinic site) lyase (Apex1), Myocyte-specific enhancer factor 2D (Mef2d) and Pre B-cell leukemia transcription factor 3 (Pbx3) was further verified in pluripotent stem cell-derived cardiomyocytes. This research offers a new understanding of cardiomyocytes in response to H2O2-induced oxidative stress and reveals additional potential therapeutic targets. The combination of two parallel omics datasets (corresponding to the transcriptome and proteome) can reduce the noise in high-throughput data and reveal the fundamental changes of the biological process, making it suitable and reliable for investigation of critical targets in many other complicated pathological processes.

Keywords: Cardiomyocyte; Melatonin; Oxidative stress; RNA-seq; Transcription factor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Apoptosis / genetics
  • Cardiotonic Agents / pharmacology*
  • Cell Line
  • DNA Repair / drug effects*
  • DNA-(Apurinic or Apyrimidinic Site) Lyase / genetics
  • DNA-(Apurinic or Apyrimidinic Site) Lyase / metabolism
  • Drugs, Chinese Herbal / pharmacology*
  • Gene Expression Profiling
  • Gene Regulatory Networks
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • Hydrogen Peroxide / antagonists & inhibitors*
  • Hydrogen Peroxide / pharmacology
  • MEF2 Transcription Factors / genetics
  • MEF2 Transcription Factors / metabolism
  • Melatonin / pharmacology
  • Microarray Analysis
  • Myocytes, Cardiac / cytology
  • Myocytes, Cardiac / drug effects*
  • Myocytes, Cardiac / metabolism
  • Oxidative Stress
  • Protein Binding
  • Proto-Oncogene Proteins / genetics
  • Proto-Oncogene Proteins / metabolism
  • Rats
  • Response Elements
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Transcriptome*

Substances

  • Cardiotonic Agents
  • Drugs, Chinese Herbal
  • Homeodomain Proteins
  • MEF2 Transcription Factors
  • MEF2D protein, human
  • Proto-Oncogene Proteins
  • Transcription Factors
  • danhong
  • proto-oncogene protein Pbx3
  • Hydrogen Peroxide
  • APEX1 protein, human
  • DNA-(Apurinic or Apyrimidinic Site) Lyase
  • Melatonin