Cellular and Molecular Biology

Antisense oligonucleotides to stromelysin mRNA inhibit injury-induced proliferation of arterial smooth muscle cells

C. Lövdahl1, J. Thyberg2, B. Cercek3, K. Blomgren2, P. Dimayuga3, B. Kallin4 and A. Hultgårdh-Nilsson1

1Department of Cell and Molecular Biology, Lund University, 2Department of Cell and Molecular Biology, Karolinska Institutet, 3Division of Cardiology, Cedars-Sinai Medical Center, UCLA School of Medicine, Los Angeles, California, USA and 4King Gustaf V Research Institute, Karolinska Hospital, Stockholm, Sweden

Offprint requests to: Anna Hultgårdh-Nilsson, Lund University, Department of Cell and Molecular Biology, Division of Connective Tissue Biology, P. O. Box 94, S-221 00 Lund, Sweden. Fax: +46-46-211 3417. e-mail: Anna.Hultgardh@medkem.lu.se


Summary. Smooth muscle cell migration and proliferation are important events in the formation of intimal lesions associated with atherosclerosis and restenosis following balloon angioplasty. To make this possible, the smooth muscle cell has to change from a contractile to an activated repair cell with capacity to synthesize DNA and extracellular matrix components. There is now considerable evidence that the extracellular matrix has important functions in modulating the phenotypic properties of smooth muscle cells, but less is known about the role of the matrix metalloproteinases. The present study investigates the role of stromelysin in the modulation of rat aortic smooth muscle cell morphology and function following mechanical injury in vitro and in vivo. Antisense mRNA oligonucleotides were used to investigate the role of stromelysin expression in injury-induced phenotypic modulation and the subsequent migration and proliferation of vascular smooth muscle cells. Cultured rat aortic smooth muscle cells and balloon-injured rat carotid arteries were used as experimental models. Light- and electron microscopy were used to follow changes in smooth muscle cell phenotype and lesion formation and incorporation of 3H-thymidine to detect DNA synthesis. Injury-induced DNA synthesis and migration in vitro were inhibited by 72% and 36%, respectively, by adding stromelysin antisense oligonucleotides to the medium prior to injury. In primary cultures, 67% of the smooth muscle cells treated with stromelysin antisense were retained in a contractile phenotype as judged by analysis of cell fine structure, compared to 15% untreated cells and 40% in cells treated with mismatched oligonucleotides. Examination of the carotid arteries one week after balloon injury likewise demonstrated a larger fraction of contractile cells in the inner parts of the media in vessels treated with antisense oligonucleotides compared to those treated with mismatched oligonucleotides. The neointima was also distinctly thinner in antisense-treated than in mismatched-treated and control arteries at this time. These findings indicate that stromelysin mRNA antisense oligonucleotides inhibited phenotypic modulation of rat arterial smooth muscle cells and so caused a decrease in migration and proliferation and neointima formation in response to vessel wall injury. Histol. Histopathol. 14, 1101-1112 (1999)

Key words: Balloon-injury, Stromelysin, Phenotypic modulation, Restenosis, Rat, Smooth muscle cells

DOI: 10.14670/HH-14.1101