Immunochemical quantification of crossline as a fluorescent advanced glycation endproduct in erythrocyte membrane proteins from diabetic patients with or without retinopathy

• Published in: Diabetic medicine Vol. 15; no. 6; pp. 458 - 462
• Main Authors: Yamaguchi, M., Nakamura, N., Nakano, K., Kitagawa, Y., Shigeta, H., Hasegawa, G., Ienaga, K., Nakamura, K., Nakazawa, Y., Fukui, I., Obayashi, H., Kondo, M.
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01.06.1998; Blackwell
• Subjects: advanced glycation endproducts; Aged; Biological and medical sciences; Blood Proteins - analysis; Case-Control Studies; crossline; Diabetes Mellitus, Type 2 - blood; Diabetes. Impaired glucose tolerance; diabetic complication; diabetic retinopathy; Diabetic Retinopathy - blood; Endocrine pancreas. Apud cells (diseases); Endocrinopathies; Enzyme-Linked Immunosorbent Assay; erythrocyte membrane; Erythrocyte Membrane - chemistry; Etiopathogenesis. Screening. Investigations. Target tissue resistance; Female; Glycation End Products, Advanced - blood; Humans; Maillard Reaction; Male; Medical sciences; Membrane Proteins - analysis; Middle Aged; Endocrinopathy; Human; Eye disease; Membrane protein; Retinopathy; Red blood cell; Complication; Immunochemistry; Non insulin dependent diabetes; ELISA assay; Glycation; Quantitative analysis
• ISSN: 0742-3071; 1096-9136; 1464-5491
• DOI: 10.1002/(SICI)1096-9136(199806)15:6<458::AID-DIA601>3.0.CO;2-Q

Crossline is a novel advanced glycation endproduct (AGE) which has both a crosslink and fluorescence similar to AGE‐protein in vivo. To assess the association of AGEs to the development of diabetic retinopathy we developed a sensitive and specific enzyme‐linked immunosorbent assay (ELISA) for crossline in blood samples and investigated the association of the development of retinopathy and erythrocyte membrane protein (EMP)‐crossline concentrations in patients with Type 2 diabetes mellitus (Type 2 DM). Crossline formation in EMP exceeded that in haemoglobin and was detectable in normal EMP samples without pretreatment by this ELISA system. Mean (±SE) EMP crossline levels were elevated 1.6‐fold in diabetic patients without retinopathy (7.6 ± 0.5 pmol mg−1, p < 0.005), 2.2‐fold in diabetic patients with non‐proliferative retinopathy (10.5 ± 0.6 pmol mg−1, p < 0.001) and 2.6‐fold in diabetic patients with proliferative retinopathy (12.0 ± 0.6 pmol mg−1, p < 0.001) compared with healthy control subjects (4.7 ± 0.5 pmol mg−1). Type 2 DM patients with retinopathy had significantly higher EMP‐crossline levels than those without retinopathy (p < 0.005). Our data suggest that elevated EMP‐crossline concentrations are associated with the presence of retinopathy in patients with Type 2 DM and EMP‐crossline measured by our ELISA may provide a useful marker for assessing the role of glycation in the development of diabetic retinopathy. © 1998 John Wiley & Sons, Ltd.