Nvar epsilon-(carboxymethyl)lysine in atherosclerotic vascular lesions as a marker for local oxidative stress Original Research Article

Previous studies suggested that Nvar epsilon-(carboxymethyl)lysine (CML), as the major product of oxidative degradation of glycated proteins and unsaturated fatty acids, represents an integrative biomarker for oxidative stress. In the present study, the level of CML in morphologically normal as well as atherosclerotic vessel walls are immunohistochemically analyzed and the in vitro formation of CML determined from glycoxidation and lipid peroxidation processes. The analysis revealed negative staining results in normal arterial walls of fetal, juvenile and young adult origin. A minor positive staining was seen in normal arteries from adults between 40 and 60 years of age with a rise in the CML-staining further increasing with rising individual age. This staining was mainly restricted to the intimal extracellular matrix and there was no intracellular staining. In arteriosclerotic vessels, in contrast, the extracellular CML-staining was significantly increased by approximately 3-fold also affecting the vascular media and adventitia. A strong intracellular staining was seen in macrophages. The degree of CML-staining correlated with the extent of the atherosclerotic changes. The in vitro studies showed a slow formation of CML of glycated proteins under aerobic conditions. No CML was formed under anaerobic conditions. Unsaturated fatty acids revealed a much faster formation of CML which reached high levels. The addition of vitamin E did not substantially suppress the CML-formation. The data suggest that the endogenous biomarker CML for oxidative stress accumulates slowly in normal arterial walls. This process is significantly increased in atherosclerosis. While the accumulation of CML in the extracellular matrix seemed to be the result of ongoing glycoxidation, the significant intracellular CML-formation in macrophages may have resulted from lipid peroxidation.