SILAC-based proteomic analysis reveals that salidroside antagonizes cobalt chloride-induced hypoxic effects by restoring the tricarboxylic acid cycle in cardiomyocytes

J Proteomics. 2016 Jan 1:130:211-20. doi: 10.1016/j.jprot.2015.09.028. Epub 2015 Oct 3.

Abstract

Hypoxic status alters the energy metabolism and induces cell injury in cardiomyocytes, and it further triggers the occurrence and development of cardiovascular diseases. Our previous studies have shown that salidroside (SAL) exhibits anti-hypoxic activity. However, the mechanisms remain obscure. In the present study, we successfully screened 92 different expression proteins in CoCl2-induced hypoxic conditions, 106 different expression proteins in the SAL-mediated anti-hypoxic group were compared with the hypoxic group using quantitative proteomics strategy, respectively. We confirmed that SAL showed a positive protective function involving the acetyl-CoA metabolic, tricarboxylic acid (TCA) cycle using bioinformatics analysis. We also demonstrated that SAL plays a critical role in restoring the TCA cycle and in protecting cardiomyocytes from oxidative injury via up-regulation expressions of PDHE1-B, ACO2, SUCLG1, SUCLG2 and down-regulation of MDH2. SAL also inhibited H9c2 cell apoptosis by inhibiting the activation of pro-apoptotic molecules caspase 3 and caspase 9 as well as activation of the anti-apoptotic molecular Bcl-2. Additionally, SAL also improved mitochondrial membrane potential (ΔΨm), reduced reactive oxygen species (ROS) and intercellular Ca(2+) concentration ([Ca(2+)]i) accumulation and inhibited the excessive consumption of ATP in H9c2 cells.

Keywords: Cobalt chloride; Hypoxic; Quantitative proteomics; SILAC; Salidroside.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / chemistry
  • Apoptosis
  • Calcium / chemistry
  • Caspase 3 / metabolism
  • Caspase 9 / metabolism
  • Cell Line
  • Chromatography, Liquid
  • Citric Acid Cycle
  • Cobalt / chemistry*
  • Computational Biology
  • Glucosides / chemistry*
  • Hypoxia / pathology
  • Membrane Potentials
  • Myocytes, Cardiac / metabolism*
  • Oxygen / chemistry
  • Phenols / chemistry*
  • Plant Extracts / chemistry
  • Proteome
  • Proteomics / methods*
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Reactive Oxygen Species / metabolism
  • Rhodiola / chemistry
  • Tandem Mass Spectrometry
  • Tricarboxylic Acids / chemistry*

Substances

  • Glucosides
  • Phenols
  • Plant Extracts
  • Proteome
  • Proto-Oncogene Proteins c-bcl-2
  • Reactive Oxygen Species
  • Tricarboxylic Acids
  • Cobalt
  • Adenosine Triphosphate
  • Caspase 3
  • Caspase 9
  • cobaltous chloride
  • rhodioloside
  • Oxygen
  • Calcium