Effect of microRNA-145 to prevent vein graft disease in rabbits by regulation of smooth muscle cell phenotype

J Thorac Cardiovasc Surg. 2014 Aug;148(2):676-82.e2. doi: 10.1016/j.jtcvs.2013.11.054. Epub 2013 Dec 31.

Abstract

Objective: Because microRNA-145 (miR-145) is a specific mediator in the regulation of the proliferation and differentiation of smooth muscle cells, we investigated the effect of miR-145 on the intimal hyperplasia in the rabbit model of vein graft disease using electroporation-mediated gene transfer.

Methods: The right jugular vein of male Japanese white rabbits was harvested and transduced with miR-145-encoding plasmids using an electroporator and then interposed in the carotid artery. At 2 or 4 weeks postoperatively, the venous graft was explanted, and the intimal thickness and intima/media area ratio were evaluated. Furthermore, 3 days after implantation, the myocardin and serum response factors were measured using real-time polymerase chain reaction. At 2 weeks after implantation, immunohistochemical investigations using mature smooth muscle markers, myosin heavy chain smooth muscle-1 and -2, and proliferation marker Ki-67 were performed.

Results: MiR-145 transduction significantly reduced the neointimal thickness at both 2 and 4 weeks (2 weeks, 52.1 ± 15.7 vs 113.2 ± 26.9 μm, P < .05, n = 6; 4 weeks, 42.4 ± 4.8 vs 136.5 ± 38.3 μm, P < .05, n = 8), and it also significantly reduced the intima/media area ratio at 4 weeks (0.22 ± 0.04 vs 1.13 ± 0.23, P < .01, n = 8). Additionally, it upregulated the mRNA expression level of myocardin compared with that in the grafts that did not receive gene transfer. Smooth muscle-2 and Ki-67 expression revealed that miR-145 transduced grafts contained more smooth muscle-2-positive mature smooth muscle cells and fewer Ki-67-positive proliferating cells.

Conclusions: Nonviral transduction of miR-145 into the bypass graft could be a novel option for preventing intimal hyperplasia in vein graft disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers / metabolism
  • Carotid Artery, Common / surgery
  • Cell Differentiation
  • Cell Proliferation
  • Disease Models, Animal
  • Electroporation
  • Gene Expression Regulation
  • Genetic Therapy*
  • Graft Occlusion, Vascular / genetics
  • Graft Occlusion, Vascular / metabolism
  • Graft Occlusion, Vascular / pathology
  • Graft Occlusion, Vascular / prevention & control*
  • Humans
  • Hyperplasia
  • Immunohistochemistry
  • Jugular Veins / metabolism
  • Jugular Veins / pathology
  • Jugular Veins / transplantation*
  • Male
  • Muscle, Smooth, Vascular / metabolism
  • Muscle, Smooth, Vascular / pathology
  • Muscle, Smooth, Vascular / transplantation*
  • Myocytes, Smooth Muscle / metabolism
  • Myocytes, Smooth Muscle / pathology
  • Myocytes, Smooth Muscle / transplantation*
  • Neointima
  • Phenotype
  • Rabbits
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Time Factors
  • Transduction, Genetic / methods

Substances

  • Biomarkers