Combined immunoelectron microscopic and computer-assisted image analyses to detect advanced glycation end-products in human myocardium

摘要: Advanced glycation end-products (AGEs) result from oxidation–reduction reactions that ensue when a sugar becomes adducted to protein. AGEs cause various complications of diabetes mellitus (DM). Experimental and clinical evidence suggest also contribute the hypertension (HTN). Little is known about abundance localization in human myocardium. In few light microscopic studies, AGE carboxymethyl lysine (CML) has been immunolabeled localized virtually exclusively walls small arteries. To more precisely delineate CML, we developed an immunoelectron (IEM) detection method using anti-CML monoclonal antibody 6D12 conjunction with computer-assisted image analysis. Antibody was pre-absorbed purified AGE-bovine serum albumin assure specificity. Antigen–antibody (ag–ab) complexes were individually identified protein A-conjugated colloidal gold counted automated system. We applied this 21 patients (pts) undergoing epicardial biopsy during coronary bypass grafting (CBG) [20 M, 1 F; mean age 65 ± 7.4 (± SEM) years]. Seven pts had neither DM nor HTN, seven + HTN. contrast prior detected CML scattered throughout cardiomyocyte all pts, but widely varying amounts. Ag–ab abundant sections through myofilaments (mean count 23.6 9.2 per μm2, range 9.4–48) even so mitochondria 34.4 11.9 14.1–68.2, P < 0.001 vs. myofilaments). vascular endothelial cells. There no statistically significant differences based on presence or absence HTN DM. conclusion, our IEM first provide detailed delineation very prevalent CBG suggesting could play role abnormal function, including altered energy metabolism.