fructosyl-lysine and Diabetes-Mellitus--Type-2

OBJECTIVE Salsalate is a nonacetylated salicylate that lowers glucose levels in people with type 2 diabetes (T2D). Here we examined whether salsalate also lowered serum-protein-bound levels of early and advanced glycation end products (AGEs) that have been implicated in diabetic vascular complications. RESEARCH DESIGN AND METHODS Participants were from the Targeting Inflammation Using Salsalate for Type 2 Diabetes (TINSAL-T2D) study, which examined the impact of salsalate treatment on hemoglobin A1c (HbA1c) and a wide variety of other parameters. One hundred eighteen participants received salsalate, 3.5 g/day for 48 weeks, and 109 received placebo. Early glycation product levels (HbA1c and fructoselysine [measured as furosine]) and AGE levels (glyoxal and methylglyoxal hydroimidazolones [G-(1)H, MG-(1)H], carboxymethyllysine [CML], carboxyethyllysine [CEL], pentosidine) were measured in patient serum samples. RESULTS Forty-eight weeks of salsalate treatment lowered levels of HbA1c and serum furosine (P < 0.001) and CML compared with placebo. The AGEs CEL and G-(1)H and MG-(1)H levels were unchanged, whereas pentosidine levels increased more than twofold (P < 0.001). Among salsalate users, increases in adiponectin levels were associated with lower HbA1c levels during follow-up (P < 0.001). Changes in renal and inflammation factor levels were not associated with changes in levels of early or late glycation factors. Pentosidine level changes were unrelated to changes in levels of renal function, inflammation, or cytokines. CONCLUSIONS Salsalate therapy was associated with a reduction in early but not late glycation end products. There was a paradoxical increase in serum pentosidine levels suggestive of an increase in oxidative stress or decreased clearance of pentosidine precursor.

Topics: Adiponectin; Adult; Anti-Inflammatory Agents, Non-Steroidal; Arginine; Diabetes Mellitus, Type 2; Diabetic Angiopathies; Female; Glycated Hemoglobin; Glycation End Products, Advanced; Humans; Lysine; Male; Middle Aged; Pyruvaldehyde; Salicylates

Complement activation via the MBL pathway has been proposed to play a role in the pathogenesis of diabetic complications. As protein glycation is increased in diabetes, we tested the possibility that the glycation product fructoselysine is a ligand for MBL and that its interaction with this protein may initiate complement activation.. We investigated the binding of MBL to fructoselysine by chromatography of human serum on fructoselysine-Sepharose, followed by Western blot and mass spectrometry analysis. We also performed enzyme-linked immunosorbent assays using purified MBL and fructoselysine-derivatized (binding assay) or mannan-coated plates (inhibition assay). Complement activation was determined by the fixation of C3d following incubation of fructoselysine-derivatized plates with serum from subjects with different levels of MBL.. MBL and its associated proteases were selectively purified from serum by chromatography on fructoselysine-Sepharose. Competition experiments indicated that MBL had a similar affinity for mannose, fructose and fructoselysine. MBL bound, in a highly cooperative manner, to fructoselysine-derivatized plates. This binding was associated with complement activation and was much lower with serum from subjects with low-MBL genotypes.. MBL binding to fructoselysine and the ensuing complement activation may provide a physiopathological link between enhanced glycation and complement activation in diabetes. The cooperative character of this binding may explain the high sensitivity of diabetic complications to hyperglycaemia.

Topics: Animals; Blood Glucose; Chromatography, Affinity; Complement Activation; Diabetes Mellitus, Type 2; Enzyme-Linked Immunosorbent Assay; Humans; Lysine; Mannose-Binding Lectin; Phosphotransferases (Alcohol Group Acceptor)

Pentosidine is an advanced glycation end product and its formation is shown to be closely related to oxidative processes. Recent studies have shown that pentosidine levels are increased not only in plasma and matrix proteins from diabetic patients, but also markedly in nondiabetic hemodialysis patients. Currently, the mechanism of accumulation and kinetics of pentosidine formation in hemodialysis patients remain unknown. Gel filtration of uremic plasma revealed that plasma pentosidine exists in the albumin fraction (approximately 90%) and, interestingly, in free form (approximately 5%) as well. Plasma free pentosidine was undetectable in subjects with normal renal function. There was a significant correlation between the plasma levels of albumin-linked and free pentosidine in hemodialysis patients. Kinetic studies indicated that dietary pentosidine was absorbed into the circulation and that, after either oral or intravenous administration of pentosidine to intact or nephrectomized rats, the plasma free pentosidine level was closely linked to the level of renal function. These findings demonstrate that: (1) Pentosidine accumulates as albumin-linked and in free form in the circulation of uremic patients; (2) dietary pentosidine can be absorbed into the circulation, thus being one possible origin of circulating free pentosidine; (3) free pentosidine may accumulate as a result of decreased glomerular filtration; and (4) the mechanism of accumulation of albumin-linked pentosidine is not related to high glucose levels. It suggests the simultaneous accumulation, during renal failure, of either unknown pentosidine precursor(s) or catalyst(s) of glycoxidation, independent of glucose.

Topics: Animals; Arginine; beta 2-Microglobulin; Creatinine; Diabetes Mellitus, Type 2; Diabetic Nephropathies; Diet; Glomerulonephritis; Humans; Kidney Failure, Chronic; Lysine; Maillard Reaction; Nephrectomy; Oxidation-Reduction; Protein Binding; Rats; Rats, Wistar; Serum Albumin; Uremia

3-Deoxyglucosone (3-DG) is a reactive dicarbonyl sugar thought to be a key intermediate in the nonenzymatic polymerization and browning of proteins by glucose. 3-DG may be formed in vivo from fructose, fructose 3-phosphate, or Amadori adducts to protein, such as N epsilon-fructoselysine (FL), all of which are known to be elevated in body fluids or tissues in diabetes. Modification of proteins by 3-DG formed in vivo is thought to be limited by enzymatic reduction of 3-DG to less reactive species, such as 3-deoxyfructose (3-DF). In this study, we have measured 3-DF, as a metabolic fingerprint of 3-DG, in plasma and urine from a group of diabetic patients and control subjects. Plasma and urinary 3-DF concentrations were significantly increased in the diabetic compared with the control population (0.853 +/- 0.189 vs. 0.494 +/- 0.072 microM, P < 0.001, and 69.9 +/- 44.2 vs. 38.7 +/- 16.1 nmol/mg creatinine, P < 0.001, respectively). Plasma and urinary 3-DF concentrations correlated strongly with one another, with HbA1c (P < 0.005 in all cases), and with urinary FL (P < 0.02 and P = 0.005, respectively). The overall increase in 3-DF concentrations in plasma and urine in diabetes and their correlation with other indexes of glycemic control suggest that increased amounts of 3-DG are formed in the body during hyperglycemia in diabetes and then metabolized to 3-DF. These observations are consistent with a role for increased formation of the dicarbonyl sugar 3-DG in the accelerated browning of tissue proteins in diabetes.

Topics: Adolescent; Adult; Aged; Biomarkers; Blood Glucose; Diabetes Mellitus; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Glycated Hemoglobin; Humans; Ketoses; Lysine; Middle Aged; Reference Values; Regression Analysis

Low density lipoprotein (LDL) from Type 1 and Type 2 diabetic patients, unlike LDL from healthy subjects, caused a 1.5- to 2.5-fold increase in the cholesterol content of cells cultured from unaffected human aortic intima, i.e. possessed atherogenic potential. LDL were further divided into two subfractions by affinity chromatography on Ricinus communis agglutinin-agarose. The amount of bound LDL was significantly higher in diabetic patients as compared with healthy subjects. Bound LDLs differed from unbound ones by significantly lowered sialic acid content, i.e. were desialylated lipoproteins. Desialylated, but not sialylated LDL subfraction induced massive cholesterol accumulation in cultured cells. Desialylated LDL subfraction in diabetic patients was also characterized by a higher degree of nonenzymatic glycation as compared to LDL subfraction with normal sialic acid level. Desialylated LDL had significantly decreased levels of free and esterified cholesterol, triglycerides and phosphatidylcholine and elevated amounts of lysophosphatidylcholine. These disturbances in lipid constituent of LDL were weak in healthy subjects, but conspicuous in diabetic patients. The results of this study have shown that there is in vivo modified LDL subfraction in the blood of diabetic patients which is characterized by crucial changes in protein and lipid moiety and are able to induce massive cholesterol accumulation in cultured cells.

Topics: Cells, Cultured; Cholesterol; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Humans; Lipids; Lipoproteins, LDL; Lysine; N-Acetylneuraminic Acid; Phospholipids; Sialic Acids

Glycation, oxidation, and browning of proteins have all been implicated in the development of diabetic complications. We measured the initial Amadori adduct, fructoselysine (FL); two Maillard products, N epsilon-(carboxymethyl) lysine (CML) and pentosidine; and fluorescence (excitation = 328 nm, emission = 378 nm) in skin collagen from 39 type 1 diabetic patients (aged 41.5 +/- 15.3 [17-73] yr; duration of diabetes 17.9 +/- 11.5 [0-46] yr, [mean +/- SD, range]). The measurements were related to the presence of background (n = 9) or proliferative (n = 16) retinopathy; early nephropathy (24-h albumin excretion rate [AER24] > or = 20 micrograms/min; n = 9); and limited joint mobility (LJM; n = 20). FL, CML, pentosidine, and fluorescence increased progressively across diabetic retinopathy (P < 0.05, P < 0.001, P < 0.05, P < 0.01, respectively). FL, CML, pentosidine, and fluorescence were also elevated in patients with early nephropathy (P < 0.05, P < 0.001, P < 0.01, P < 0.01, respectively). There was no association with LJM. Controlling for age, sex, and duration of diabetes using logistic regression, FL and CML were independently associated with retinopathy (FL odds ratio (OR) = 1.06, 95% confidence interval (CI) = 1.01-1.12, P < 0.05; CML OR = 6.77, 95% CI = 1.33-34.56, P < 0.05) and with early nephropathy (FL OR = 1.05, 95% CI = 1.01-1.10, P < 0.05; CML OR = 13.44, 95% CI = 2.00-93.30, P < 0.01). The associations between fluorescence and retinopathy and between pentosidine and nephropathy approached significance (P = 0.05). These data show that FL and Maillard products in skin correlate with functional abnormalities in other tissues and suggest that protein glycation and oxidation (glycoxidation) may be implicated in the development of diabetic retinopathy and early nephropathy.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Arginine; Collagen; Diabetes Mellitus, Type 2; Diabetic Angiopathies; Diabetic Nephropathies; Diabetic Retinopathy; Female; Humans; Lysine; Maillard Reaction; Male; Microcirculation; Middle Aged; Skin

Furosine, which was formed by acid hydrolysis of fructose-lysine, was determined and used as an indicator of glycosylated protein. The diabetic patients had significantly higher fructose-lysine levels in finger nails than healthy subjects [10.8 +/- 4.6% (mean +/- S.D.) vs 4.2 +/- 1.1%]. The best correlation was found between the fructose-lysine value and the fasting blood glucose level determined 3 to 5 months before sampling nails in diabetics. These results suggest that the furosine derived from fructose-lysine in finger nails may become an indicator of blood glucose control during the past 3 to 5 months in diabetics.

Topics: Adult; Aged; Aged, 80 and over; Blood Glucose; Diabetes Mellitus, Type 2; Female; Humans; Lysine; Male; Middle Aged; Nails

Fructose-lysine, which is formed by binding glucose to lysine, is changed by acid hydrolysis into furosine. Furosine derived from fructose-lysine of hair was determined by high-performance liquid chromatography according to the method of Schleicher et al. Furosine values were significantly higher in diabetic patients than in healthy subjects, and significantly correlated with the stable components of hemoglobin A1 (HbA1) values. These results suggest that furosine, like HbA1, may become an indicator of past blood glucose control at any time in diabetic patients and be useful in investigating diabetic complications on the level of tissue.

Topics: Adult; Aged; Chromatography, High Pressure Liquid; Diabetes Mellitus, Type 2; Female; Glycated Hemoglobin; Hair; Humans; Lysine; Male; Middle Aged; Proteins