TY - JOUR
T1 - A quick, simple method for detecting circulating fluorescent advanced glycation end-products: Correlation with in vitro and in vivo non-enzymatic glycation
AU - Villa, M.
AU - Parravano, M.
AU - Micheli, A.
AU - Gaddini, L.
AU - Matteucci, A.
AU - Mallozzi, C.
AU - Facchiano, F.
AU - Malchiodi-Albedi, F.
AU - Pricci, F.
N1 - Export Date: 4 April 2018
CODEN: METAA
Correspondence Address: Pricci, F.; Dept of Cardiovascular, Dysmetabolic and Aging-associated diseasesUnited States; email: flavia.pricci@iss.it
Chemicals/CAS: glucose, 50-99-7, 84778-64-3; Fluorescent Dyes; Glycosylation End Products, Advanced; Serum Albumin, Bovine
References: Giacco, F., Brownlee, M., Oxidative stress and diabetic complications (2010) Circ Res, 107 (9), pp. 1058-1070; Takeuchi, M., Kikuchi, S., Sasaki, N., Suzuki, T., Watai, T., Iwaki, M., Involvement of advanced glycation end-products (AGEs) in Alzheimer's disease (2004) Curr Alzheimer Res, 1, pp. 39-46; Baynes, J.W., The role of AGEs in aging: causation or correlation (2001) Exp Gerontol, 36, pp. 1527-1537; Brownlee, M., Biochemistry and molecular cell biology of diabetic complications (2001) Nature, 414 (6865), pp. 813-820; Singh, R., Barden, A., Mori, T., Beilin, L., Advanced glycation end-products: a review (2001) Diabetologia, 44, pp. 129-146; Di Mario, U., Pugliese, G., 15th Golgi lecture: from hyperglycaemia to the dysregulation of vascular remodelling in diabetes (2001) Diabetologia, 44, pp. 674-692; Ahmed, N., Advanced glycation endproducts-role in pathology of diabetic complications (2005) Diabetes Res Clin Pract, 67 (1), pp. 3-21; de Oliveira, S.C., Delbosc, S., Araïs, C., Monnier, L., Cristol, J.P., Pares-Herbute, N., Modulation of CD36 protein expression by AGEs and insulin in aortic VSMCs from diabetic and non-diabetic rats (2008) Nutr Metab Cardiovasc Dis, 18 (1), pp. 23-30; Pricci, F., Leto, G., Amadio, L., Iacobini, C., Cordone, S., Catalano, S., Oxidative stress in diabetes-induced endothelial dysfunction involvement of nitric oxide and protein kinase C (2003) Free Radic Biol Med, 35 (6), pp. 683-694; Pinzón-Duarte, G., Kohler, K., Arango-González, B., Guenther, E., Cell differentiation, synaptogenesis, and influence of the retinal pigment epithelium in a rat neonatal organotypic retina culture (2000) Vision Res, 40 (25), pp. 3455-3465; Sampathkumar, R., Balasubramanyam, M., Rema, M., Premanand, C., Mohan, V., A novel advanced glycation index and its association with diabetes and microangiopathy (2005) Metabolism, 54 (8), pp. 1002-1007; Zong, H., Ward, M., Stitt, A.W., AGEs, RAGE, and diabetic retinopathy (2011) Curr Diab Rep, 11 (4), pp. 244-252; Cohen, M.P., Hud, E., Shea, E., Shearman, C.W., Vitreous fluid of db/db mice exhibits alterations in angiogenic and metabolic factors consistent with early diabetic retinopathy (2008) Ophthalmic Res, 40 (1), pp. 5-9; Géhl, Z., Bakondi, E., Resch, M.D., Hegedűs, C., Kovács, K., Lakatos, P., Diabetes-induced oxidative stress in the vitreous humor (2016) Redox Biol, 9, pp. 100-103
PY - 2017
Y1 - 2017
N2 - Objective Advanced glycation end-products (AGEs) constitute a highly heterogeneous family of compounds, relevant in the pathogenesis of diabetic complications, which could represent efficient biomarkers of disease progression and drug response. Unfortunately, due to their chemical heterogeneity, no method has been validated to faithfully monitor their levels in the course of the disease. In this study, we refine a procedure to quantitatively analyze fluorescent AGEs (fAGEs), a subset considered remarkably representative of the entire AGE family, and measure them in in vitro glycated BSA (gBSA) and in plasma and vitreous of diabetic rats, for testing its use to possibly quantify circulating AGEs in patients, as markers of metabolic control. Methods fAGE levels were evaluated by spectrofluorimetric analysis in in vitro and in vivo experimental models. BSA was glycated in vitro with increasing D-glucose concentrations for a fixed time or with a fixed D-glucose concentration for increasing time. In in vivo experiments, streptozotocin-induced diabetic rats were studied at 1, 3, 6 and 12 weeks to analyze plasma and vitreous. To confirm the presence of AGEs in our models, non-diabetic rat retinal explants were exposed to high glucose (HG), to reproduce short-term effects, or in vitro gBSA, to reproduce long-term effects of elevated glucose concentrations. Rat retinal explants and diabetic retinal tissues were evaluated for the receptor for advanced glycation end-product (RAGE) by Western blot analysis. Results In in vitro experiments, fluorescence emission showed glucose concentration- and time-dependent increase of fAGEs in gBSA (p ≤ 0.05). In streptozotocin-induced diabetic rats, fAGE in plasma and vitrei showed an increase at 6 (p ≤ 0.005) and 12 (p ≤ 0.05) weeks of diabetes, with respect to control. RAGE was time-dependently upregulated in retinas incubated with gBSA, but not with HG, and in diabetic retinal tissue, substantiating exposure to AGEs. Conclusions Applying the proposed technique, we could show that fAGEs levels increase with glucose concentration and time of exposure in vitro. Furthermore, in diabetic rats, it showed that circulating fAGEs are similarly upregulated as those in vitreous, suggesting a correlation between circulating and tissue AGEs. These results support the use of this method as a simple and reliable test to measure circulating fAGEs and monitor diabetes progression. © 2017 Elsevier Inc.
AB - Objective Advanced glycation end-products (AGEs) constitute a highly heterogeneous family of compounds, relevant in the pathogenesis of diabetic complications, which could represent efficient biomarkers of disease progression and drug response. Unfortunately, due to their chemical heterogeneity, no method has been validated to faithfully monitor their levels in the course of the disease. In this study, we refine a procedure to quantitatively analyze fluorescent AGEs (fAGEs), a subset considered remarkably representative of the entire AGE family, and measure them in in vitro glycated BSA (gBSA) and in plasma and vitreous of diabetic rats, for testing its use to possibly quantify circulating AGEs in patients, as markers of metabolic control. Methods fAGE levels were evaluated by spectrofluorimetric analysis in in vitro and in vivo experimental models. BSA was glycated in vitro with increasing D-glucose concentrations for a fixed time or with a fixed D-glucose concentration for increasing time. In in vivo experiments, streptozotocin-induced diabetic rats were studied at 1, 3, 6 and 12 weeks to analyze plasma and vitreous. To confirm the presence of AGEs in our models, non-diabetic rat retinal explants were exposed to high glucose (HG), to reproduce short-term effects, or in vitro gBSA, to reproduce long-term effects of elevated glucose concentrations. Rat retinal explants and diabetic retinal tissues were evaluated for the receptor for advanced glycation end-product (RAGE) by Western blot analysis. Results In in vitro experiments, fluorescence emission showed glucose concentration- and time-dependent increase of fAGEs in gBSA (p ≤ 0.05). In streptozotocin-induced diabetic rats, fAGE in plasma and vitrei showed an increase at 6 (p ≤ 0.005) and 12 (p ≤ 0.05) weeks of diabetes, with respect to control. RAGE was time-dependently upregulated in retinas incubated with gBSA, but not with HG, and in diabetic retinal tissue, substantiating exposure to AGEs. Conclusions Applying the proposed technique, we could show that fAGEs levels increase with glucose concentration and time of exposure in vitro. Furthermore, in diabetic rats, it showed that circulating fAGEs are similarly upregulated as those in vitreous, suggesting a correlation between circulating and tissue AGEs. These results support the use of this method as a simple and reliable test to measure circulating fAGEs and monitor diabetes progression. © 2017 Elsevier Inc.
KW - Advanced glycation end-product
KW - Diabetes
KW - Plasma
KW - Rat
KW - Vitreous
KW - advanced glycation end product
KW - glucose
KW - bovine serum albumin
KW - fluorescent dye
KW - animal experiment
KW - animal model
KW - animal tissue
KW - Article
KW - controlled study
KW - glycation
KW - in vitro study
KW - in vivo study
KW - nonhuman
KW - priority journal
KW - rat
KW - retina tissue
KW - spectrofluorometry
KW - streptozotocin-induced diabetes mellitus
KW - vitreous body
KW - Western blotting
KW - animal
KW - blood
KW - chemistry
KW - diabetic retinopathy
KW - experimental diabetes mellitus
KW - male
KW - metabolism
KW - pathology
KW - polyacrylamide gel electrophoresis
KW - procedures
KW - retina
KW - Sprague Dawley rat
KW - Animals
KW - Diabetes Mellitus, Experimental
KW - Diabetic Retinopathy
KW - Electrophoresis, Polyacrylamide Gel
KW - Fluorescent Dyes
KW - Glycosylation End Products, Advanced
KW - Male
KW - Rats
KW - Rats, Sprague-Dawley
KW - Retina
KW - Serum Albumin, Bovine
KW - Spectrometry, Fluorescence
U2 - 10.1016/j.metabol.2017.03.004
DO - 10.1016/j.metabol.2017.03.004
M3 - Article
VL - 71
SP - 64
EP - 69
JO - Metabolism: Clinical and Experimental
JF - Metabolism: Clinical and Experimental
SN - 0026-0495
ER -