The effect of C-reactive protein deposition on myocardium with ischaemia-reperfusion injury in rats

Interact Cardiovasc Thorac Surg. 2017 Aug 1;25(2):260-267. doi: 10.1093/icvts/ivx107.

Abstract

Objectives: We evaluated the effect of monomeric C-reactive protein (CRP) deposition on areas at risk (AAR) of myocardium with ischaemia-reperfusion injury.

Methods: Myocardial ischaemia-reperfusion injury model was produced by ligation of the left anterior descending coronary artery for 45 min followed by 45 min of reperfusion using female Sprague-Dawley rats. Tissue from non-ischaemic areas, areas at risk and infarct areas determined by Evans blue and 2,3,5-triphenyltetrazolium chloride staining was obtained from the sham group, the ischaemia-reperfusion injury without C-reactive protein (CRP) injection group (I/R only group), and the ischaemia-reperfusion injury with CRP injection group (I/R + CRP group). We assessed the effect of CRP injection on infarct size, CRP deposition, CRP and IL-6 mRNA expression, the third component of complement (C3) immunodeposition and mitochondrial structural remodelling with apoptosis by quantitative RT-PCR analyses, immunohistochemistry, direct immunofluorescence, electron microscopy and Terminal deoxynucleotide transferase dUTP Nick End Labelling assay, respectively. All images were analysed using an automated morphology tool.

Results: The infarct area significantly increased in the I/R + CRP group compared to the I/R only group. The anti CRP antibody confirmed that CRP deposition occurred in both the infarct and area at risk (AAR) of the I/R + CRP group. The myocardium did not exhibit CRP mRNA expression, and the CRP treatment group showed a tendency for IL-6 to increase without statistical significance. Activated C3, apoptosis and mitochondrial destruction increased on AAR and infarct area in the I/R + CRP group.

Conclusions: These results strongly suggest the active participation of the deposition of CRP on AAR in the progression of myocardial infarction following ischaemia-reperfusion injury, accompanied by complement activation and mitochondrial change.

Keywords: Apoptosis; C-reactive protein; Mitochondria; Myocardial infarction; Myocardial reperfusion injury.

MeSH terms

  • Animals
  • C-Reactive Protein / biosynthesis*
  • C-Reactive Protein / genetics
  • Disease Models, Animal
  • Female
  • Gene Expression Regulation
  • Immunohistochemistry
  • Interleukin-6 / biosynthesis
  • Interleukin-6 / genetics
  • Microscopy, Electron, Transmission
  • Myocardial Reperfusion Injury / genetics
  • Myocardial Reperfusion Injury / metabolism*
  • Myocardial Reperfusion Injury / pathology
  • Myocardium / metabolism*
  • Myocardium / ultrastructure
  • RNA, Messenger / genetics
  • Rats
  • Rats, Sprague-Dawley
  • Real-Time Polymerase Chain Reaction

Substances

  • Interleukin-6
  • RNA, Messenger
  • C-Reactive Protein