The Immuno-quantification of Caveolin-1 and eNOS in Human and Rabbit Diseased Blood Vessels

Anthony Zulli ^1*, Brian F. Buxton ¹, M. Jane Black ¹, Ziqiu Ming ¹, Alex Cameron ¹ and David L. Hare ¹

¹ Departments of Cardiology (AZ,ZM,AC,DLH) and Cardiac Surgery (BFB) and Medicine (AZ), University of Melbourne, Austin Health, Heidelberg, Australia, and Department of Anatomy and Cell Biology, Monash University, Clayton, Australia (MJB)

^* To whom correspondence should be addressed. E-mail: azulli{at}unimelb.edu.au.

Submitted on March 2, 2005
Accepted on 15 June 2005

	Abstract

In this study, caveolin-1 (Cav-1), an inhibitor of endothelial nitric oxide synthase (eNOS) was semi-quantified in diseased human and rabbit blood vessels. New Zealand White rabbits were fed for twelve weeks a high methionine diet (to induce intimal hyperplasia), 0.5% cholesterol diet, a normal diet or the combination of both experimental diets. Excess segments of human internal mammary arteries (IMA) and radial arteries (RA) were obtained from patients undergoing coronary artery bypass surgery. eNOS and caveolin-1 were localized throughout both human and rabbit vessels. In rabbit arteries, eNOS was significantly increased in the endothelium overlying intimal thickening and atherosclerotic plaques compared with the adjacent endothelium overlying normal media. Interestingly, the endothelial Cav-1:eNOS ratio increased 5-fold only in endothelium overlying plaques, but decreased in endothelium overlying vessels with neo-intimal thickening. In human tissue, there was no difference between RA and IMA eNOS immunoreactivity in endothelium, intima or media, however RA endothelial, intimal and medial cav-1 immunoreactivity increased 4-fold (p<0.02), 8-fold (p<0.001) and 4-fold (p<0.004) respectively compared with IMA. Furthermore, the cav-1:eNOS immunostaining ratio in the media correlated with intimal thickening (r² = 0.5). Conclusion: Our results suggest a close relationship between increased cav-1 and diseased blood vessels.

Key Words: atherosclerosis, radial artery, caveolin-1, eNOS, immunohistochemistry

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