The complementary effects of atorvastatin and exercise treatment on the composition and stability of the atherosclerotic plaques in ApoE knockout mice

PLoS One. 2014 Sep 29;9(9):e108240. doi: 10.1371/journal.pone.0108240. eCollection 2014.

Abstract

Aim: This study aimed to investigate the effects of combined atorvastatin and exercise treatment on the composition and stability of the atherosclerotic plaques in apolipoproteinE (apoE) knockout mice.

Methods: Forty male, apoE-/- mice were fed a high-fat diet for 16 weeks. Thereafter, while maintained on high-fat diet, they were randomized into four (n = 10) groups for 8 additional weeks: Group CO: Control. Group AT: Atorvastatin treatment (10 mg/Kg/day). Group EX: Exercise-training on treadmill. Group AT+EX: Atorvastatin and simultaneous exercise training. At the study's end, plasma cholesterol levels, lipids and triglycerides were measured, along with the circulating concentrations of matrix-metalloproteinases (MMP-2,3,8,9) and their inhibitors (TIMP-1,2,3). Plaque area and the relative concentrations of collagen, elastin, macrophages, smooth muscle cells, MMP-2,3,8,9 and TIMP-1,2,3 within plaques were determined. Lastly, MMP activity was assessed in the aortic arch.

Results: All intervention groups showed a lower degree of lumen stenosis, with atheromatous plaques containing more collagen and elastin. AT+EX group had less stenosis and more elastin compared to single intervention groups. MMP-3,-8 -9 and macrophage intra-plaque levels were reduced in all intervention groups. EX group had increased TIMP-1 levels within the lesions, while TIMP-2 was decreased in all intervention groups. The blood levels of the above molecules increased during atherosclerosis development, but they did not change after the therapeutic interventions in accordance to their intra-plaque levels.

Conclusion: The two therapeutic strategies act with synergy regarding the extent of the lesions and lumen stenosis. They stabilize the plaque, increasing its content in elastin and collagen, by influencing the MMP/TIMP equilibrium, which is mainly associated with the macrophage amount. While the increased MMP-2,-3,-8 -9, as well as TIMP-1 and TIMP-2 circulating levels are markers of atherosclerosis, they are not correlated with their corresponding concentrations within the lesions after the therapeutic interventions, and cannot serve as markers for the disease development/amelioration.

Trial registration: ClinicalTrials.gov NCT00636766.

Publication types

  • Clinical Trial
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apolipoproteins E / genetics*
  • Atorvastatin
  • Blood Glucose
  • Cholesterol / blood
  • Collagen / metabolism
  • Diet, High-Fat
  • Elastin / metabolism
  • Heptanoic Acids / pharmacology*
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors / pharmacology*
  • Macrophages
  • Male
  • Matrix Metalloproteinase 2 / blood
  • Matrix Metalloproteinase 3 / blood
  • Matrix Metalloproteinase 8 / blood
  • Matrix Metalloproteinase 9 / blood
  • Mice
  • Mice, Knockout
  • Myocytes, Smooth Muscle
  • Physical Conditioning, Animal / methods*
  • Plaque, Atherosclerotic / metabolism*
  • Pyrroles / pharmacology*
  • Random Allocation
  • Tissue Inhibitor of Metalloproteinase-1 / blood
  • Tissue Inhibitor of Metalloproteinase-2 / blood
  • Tissue Inhibitor of Metalloproteinase-3 / blood
  • Triglycerides / blood

Substances

  • Apolipoproteins E
  • Blood Glucose
  • Heptanoic Acids
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors
  • Pyrroles
  • Timp1 protein, mouse
  • Tissue Inhibitor of Metalloproteinase-1
  • Tissue Inhibitor of Metalloproteinase-3
  • Triglycerides
  • Tissue Inhibitor of Metalloproteinase-2
  • Collagen
  • Elastin
  • Cholesterol
  • Atorvastatin
  • Matrix Metalloproteinase 3
  • Mmp3 protein, mouse
  • Matrix Metalloproteinase 2
  • Mmp2 protein, mouse
  • MMP8 protein, mouse
  • Matrix Metalloproteinase 8
  • Matrix Metalloproteinase 9
  • Mmp9 protein, mouse

Associated data

  • ClinicalTrials.gov/NCT00636766

Grants and funding

The project was co-funded by the Operational Program “Competitiveness and Entrepreneurship” and Regional Operational Programmes of the National Strategic Reference Framework (NSRF) 2007–2013 (http://www.espa.gr/en/Pages/default.aspx). “SYNERGASIA”: “Collaborative projects of small and medium scale”. ClinicalTrials.gov Identifier: NCT00636766. NPE Kadoglou was granted by the Greek State Scholarships Foundation (http://www.iky.gr/en). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.