Measurement in vivo of irreversible degradation of low density lipoprotein in the rabbit aorta. Predominance of intimal degradation.

Abstract

The development of a highly sensitive method for assessing, tissue by tissue, the rates of irreversible protein degradation in vivo has allowed us to quantify low density lipoprotein degradation in the normal rabbit aorta and to localize it. The method depends upon the fact that tyramine-cellobiose, like sucrose used in previous studies, can be covalently attached to proteins, enter cells with them, and then remains trapped within the cell after the remainder of the protein molecule has been degraded. Rabbit LDL (d = 1.02 to d = 1.06 g/ml) was labeled with 125I-tyramine-cellobiose and injected into rabbits. Aortic 125I content 24 hours later served as a cumulative measure of degraded LDL (after appropriate corrections for any intact, nondegraded LDL present). Calculated aortic degradation of LDL averaged 9.4 X 10(-3) percent of the plasma pool per g aortic wet weight per day (n = 6). Intimal cells, obtained by gentle swabbing, accounted for fully 40% of total aortic degradation even though the intima represented less than 5% of the aortic mass. Autoradiography confirmed the high concentration of label in the intima. Degradation of unmodified and reductively methylated LDL were compared. The fractional rate of degradation of methylated LDL by the intima was 50% to 60% of that for native LDL, indicating that 40% to 50% of LDL degradation in the intima, predominantly endothelial cells, is mediated by LDL receptors.