Nogo‐B expression, in arterial intima, is impeded in the early stages of atherosclerosis in humans

Abstract

Nogo-B (Reticulon 4B) is considered to be a novel vascular marker, which may have a protective role in injury-induced neointima formation and atherosclerosis. Nogo A/B is found to be crucial for monocyte/macrophage recruitment in acute inflammation and it is expressed in CD68+macrophages. We hypothesize that macrophage infiltration in atherosclerosis is not dependent on Nogo-B expression in arterial wall. We have assessed Nogo-B expression and macrophage accumulation in the iliac arteries of healthy organ donors and organ donors with cardiovascular risk factors. Paraffin sections of 66 iliac arteries, from 44 deceased organ donors (17 women and 27 men), were studied. The healthy and cardiovascular risk (CVR) subgroups were created. With regard to staging of the atherosclerotic process, the thickness of arterial intima was measured in digitalized images of H+E stained tissue sections. Immunohistochemical reactions (Nogo-B and CD68) were carried out in all arteries (66 samples). Western blotting (WB-19 samples) and real-time PCR (27 samples) were performed on selected arteries. Significantly higher Nogo-B expression was demonstrated in the intima of the healthy subjects' subgroup, using immunohistochemistry. WB and real-time PCR revealed a trend toward lower Nogo-B expression in the adventitia of the CVR subgroup. Furthermore, the thickness of the intima was found to negatively correlate with the expression of Nogo-B in the intima and media (r=-0.32; p<0.05; r=-0.32; p<0.05). Macrophage infiltrates were more prominent in intima of CVR subjects (0.65 vs 3.52 a.u.; p<0.01). Macrophage density in intima increased with atherosclerosis progression (r=0.37; p<0.01). CD68 macrophages density in adventitia was lower in CVR arteries than in healthy arteries. The expression of Nogo-B, in arterial intima, is impeded in the early stages of atherosclerosis. Accumulation of arterial intimal CD68 macrophages has been shown to progress; however, the overall macrophage density in the adventitia is reduced in arteries shown to have intimal thickening. Macrophage infiltration is not accompanied by Nogo-B expression in atherosclerotic arteries.