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(Circulation. 2001;103:1446.)
© 2001 American Heart Association, Inc.

Basic Science Reports

Ribozyme-Mediated Inhibition of Rat Leukocyte-Type 12-Lipoxygenase Prevents Intimal Hyperplasia in Balloon-Injured Rat Carotid Arteries

Jia-Li Gu, PhD; Hong Pei, MD; Lisa Thomas, BS; Jerry L. Nadler, MD; John J. Rossi, PhD; Linda Lanting, BS; Rama Natarajan, PhD

From the Gonda Diabetes Center (J.J.R., L.L., R.N.), Beckman Research Institute of the City of Hope, Duarte, Calif; the Division of Endocrinology and Metabolism (J.-L.G., H.P., J.L.N.), University of Virginia Health Science Center, Charlottesville; and Cedars Sinai Medical Center (L.T.), Los Angeles, Calif.

Correspondence to Jerry L. Nadler, MD, Department of Internal Medicine, Division of Endocrinology and Metabolism, University of Virginia School of Medicine, MR4 Building, PO Box 801405, Charlottesville, VA, 22908. E-mail jln2n{at}virginia.edu

Background—12-Lipoxygenase (12-LO) products of arachidonate metabolism have growth and chemotactic effects in vascular smooth muscle cells. We have also recently demonstrated increased 12-LO mRNA and protein expression in the neointima of balloon-injured rat carotid arteries. In this study, we evaluated whether 12-LO activation plays a role in neointimal thickening in this rat model by using a specific ribozyme (Rz) directed to rat 12-LO.

Methods and Results—We designed a chimeric DNA-RNA hammerhead Rz to cleave rat leukocyte-type 12-LO mRNA. This Rz dose-dependently cleaved a 166-nucleotide target 12-LO mRNA substrate in vitro and reduced 12-LO mRNA and protein expression in rat vascular smooth muscle cells. A control mutant Rz (MRz) with a point mutation in the catalytic site was inactive. To test the in vivo efficacy of the 12-LO Rz, the left common carotid arteries of rats were injured with a balloon catheter. The distal half of the injured arteries was treated with Rz or MRz mixed with lipofectin. The proximal half received only lipofectin. Twelve days after injury, intima-to-media ratios were significantly lower in the Rz-treated sections than in untreated sections from the same rat (0.742±0.16 versus 1.749±0.12, P<0.001). In contrast, the MRz had no significant effect.

Conclusions—These results indicate the important role of the leukocyte-type 12-LO pathway in restenosis in response to injury.

Key Words: restenosis • lipids • vasculature • growth substances

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