Thrombin generation in type 2 diabetes with albuminuria and macrovascular disease.

BACKGROUND: Albuminuria is an indicator of cardiovascular morbidity and mortality in patients with type 2 diabetic mellitus (T2DM). MATERIALS AND METHODS: In our cross-sectional study, we measured thrombin generation (TG), a key process in haemostasis and a tool to detect an individual's coagulation potential, in normo-, micro- and macroalbuminuria in T2DM with and without macrovascular disease (MVD). The TG-assay was performed, and the TG-curve [including the lag phase, peak thrombin and area under the curve (AUC)] was analysed. RESULTS: A total of 160 patients (62 women; mean age ± SD: 67 ± 11 years) with T2DM and normo-, micro- or macroalbuminuria were investigated. Of those, 90 (56%) patients had normoalbuminuria, 40 (25%) microalbuminuria and 30 (19%) macroalbuminuria. The AUC between the groups of patients with normo-, micro- and macroalbuminuria was statistically significantly different [3297 (2785; 3764) vs. 3222 (2381; 3678) vs. 3726 (3153; 4235) nM Thrombin; P = 0AE019]. T2DM patients with MVD (n = 121) had a significantly shorter lag phase [12 (9; 16) vs. 20 (15; 25) min; P < 0AE001], a significantly higher peak thrombin [233 (130; 339) vs. 133 (82; 187) nM; P < 0AE001] and a significantly higher AUC [3464 (2969; 3868) vs. 3091 (2384; 3619) nM Thrombin; P = 0AE01] than T2DM patients without MVD (n = 39), indicating an earlier and higher thrombin generation. CONCLUSION: Our results support the hypothesis that TG may be involved in the pathogenesis of MVD in diabetic nephropathy as for the first time, we could show that patients with T2DM in different stages of diabetic nephropathy had disturbances in thrombin generation.

YKL-40 is elevated in patients with peripheral arterial disease and diabetes or pre-diabetes.

OBJECTIVE: YKL-40 is secreted by macrophages in atherosclerotic lesions and involved in plaque rupture. YKL-40 is elevated in coronary artery disease, and predicts cardiovascular mortality. Experimental in vivo and in vitro data suggest a role of YKL-40 in tissue remodeling. A disease modulating potency of YKL-40 was not investigated in peripheral arterial disease (PAD). METHODS: We measured YKL-40 in 460 subjects: 316 PAD: 71 normal glucose metabolism (PAD-NGM), 90 pre-diabetes (PAD-PREDM) and 155 diabetes (PAD-DM); 20 diabetes with atherosclerosis but without PAD (AS-DM); 85 diabetes without macro-vascular complications (DM) and 39 healthy controls (CO). RESULTS: YKL-40 is higher in PAD vs. CO (median [25-75 percentile]: 103 [69-159] vs. 43 [30-80]ng/ml; p<0.001). In addition, YKL-40 is elevated in DM (p<0.001), PAD-NGM (p=0.001), PAD-PREDM (p<0.001), PAD-DM (p<0.001) and AS-DM (p=0.002) compared to CO. Among PAD, YKL-40 is increased in PAD-PREDM (p=0.001) and PAD-DM (p=0.01) vs. PAD-NGM. By multivariate regression YKL-40 is significantly associated with age (beta=0.272), triglycerides (beta=0.216), aspartate-amino-transferase (beta=0.177) and c-reactive-protein (beta=0.178). Underpinning its role YKL-40 was found to be associated with micro-/macroalbuminuria (p=0.014/p=008)--a strong remodeling inducer. In addition, YKL-40 was elevated in existence of mediasclerosis (p=0.008), a remodeling process. CONCLUSION: We are first to show that YKL-40 is higher in subjects with peripheral arterial disease. YKL-40 was higher in PAD patients with pre-/diabetes. In addition, YKL-40 was associated with the "severity" of generalized atherosclerosis estimated by affected vascular beds. All our findings point towards a role of YKL-40 in the progression/prognosis of patients with PAD and concomitant diabetes.

Fetuin-A levels are increased in patients with type 2 diabetes and peripheral arterial disease.

OBJECTIVE: Low levels of fetuin-A, a systemic calcification inhibitor, are linked to mortality in patients on dialysis. In contrast, elevated fetuin-A is associated with cardiovascular events in non-renal patients. We investigated fetuin-A in patients with type 2 diabetes and peripheral arterial disease (PAD). RESEARCH DESIGN AND METHODS: We studied fetuin-A in 76 patients with PAD and normal glucose metabolism (NGM-PAD) and in 129 patients with PAD and type 2 diabetes (type 2 diabetes-PAD). Additionally, 40 patients with diabetes without any complications (type 2 diabetes-non-PAD) were examined. RESULTS: Type 2 diabetes-PAD subjects (399 ± 155 µg/ml) had significantly higher fetuin-A levels than type 2 diabetes-non-PAD subjects (247 ± 42; P < 0.001). In NGM-PAD subjects (376 ± 144), fetuin-A was significantly higher than in type 2 diabetes-non-PAD subjects (P < 0.001). Type 2 diabetes-PAD patients with mediasclerosis had lower fetuin-A than subjects without (P < 0.03). Regression analysis in type 2 diabetes-PAD subjects revealed that glycated A1C (P < 0.001) and mediasclerosis (P = 0.004) were the strongest predictors of fetuin-A. Multivariate regression revealed that a 1-SD increase in fetuin-A was associated with an odds ratio (OR) of 2.1 (95% CI 1.1-3.3; P < 0.001) for the prevalence of PAD and an OR of 1.4 (1.0-1.7, P = 0.039) for the prevalence of myocardial infarction. CONCLUSIONS: In contrast to previous findings, fetuin-A was higher in type 2 diabetes-PAD patients than in type 2 diabetes-non-PAD patients. In NGM-PAD patients, fetuin-A was also higher than in type 2 diabetes-non-PAD patients. In type 2 diabetes-PAD patients, fetuin-A was inversely associated with mediasclerosis-the calcification process pathognomonic for diabetic PAD. This association persisted in multivariate regression, which is in line with the calcification inhibition in coronary heart or renal disease.

Circulating angiopoietic cells and diabetic retinopathy in type 2 diabetes mellitus, with or without macrovascular disease.

PURPOSE: To investigate different types of circulating angiopoietic cells, such as vasculogenic circulating progenitor cells (CPCs), endothelial progenitor cells (EPCs), and mature EPCs (matEPCs) in patients with type 2 diabetes mellitus (T2DM), with or without diabetic retinopathy (DR) and with or without macrovascular disease (MVD). METHODS: One hundred twenty-six patients with T2DM-66 with MVD and 60 without MVD-were enrolled in a case-control study. MVD comprised coronary heart disease, peripheral arterial disease, stroke, or various combinations of those conditions. By a modified Early Treatment of Diabetic Retinopathy Study (ETDRS) classification, 55 patients were classified without DR (CO), 19 with mild nonproliferative DR (mNPDR), 16 with moderate-severe NPDR (msNPDR), 19 with early proliferative diabetic retinopathy (ePDR), and 17 with high-risk PDR (hrPDR). CPCs (CD34/CD133), EPCs (CD34/CD133/CD30), and matEPCs (CD34/CD133/CD309/CD31) were enumerated by flow cytometry. RESULTS: Patients with MVD CPCs, EPCs, and matEPCs showed a significant, stepwise decline with advancing stages of retinopathy. In contrast, in the patients without MVD, EPCs and matEPCs reached up to 56% of CPCs and 37% of EPCs. On the other hand, the percentage of EPCs and matEPCs was reduced to 5% of CPCs and EPCs each in MVD patients. Thus, the percentage of EPCs and matEPCs in comparison with that of CPCs and EPCs represented an 11- and 7-fold difference. CONCLUSIONS: The circulating angiopoietic CPCs, EPCs, and matEPCs in T2DM patients with DR had a different regulations, with increasing relative differences occurring in proliferative DR, apparently depending on the macrovascular comorbidities. Patients with MVD showed a strong retinopathy-stage-dependent depletion of all angiopoietic cells.

Correlation of different circulating endothelial progenitor cells to stages of diabetic retinopathy: first in vivo data.

PURPOSE: To investigate vasculogenic circulating progenitor cells (CPCs), endothelial progenitor cells (EPCs), and mature EPCs in patients with type 1 diabetes mellitus (T1DM) with or without diabetic retinopathy (DR). METHODS: A case-control study comparing 90 patients with T1DM with and without DR was performed. Patients were studied and staged for retinopathy according to the Early Treatment of Diabetic Retinopathy Study (ETDRS) classification. Ninety patients were included: 30 without DR (control [CO]), 30 with mild nonproliferative DR (mNPDR), 10 with moderate-severe NPDR (msNPDR), 10 with mild-moderate proliferative diabetic retinopathy (mmPDR), and 10 with high-risk PDR (hrPDR). CPCs (CD34/CD133), EPCs (CD34/CD133/CD309), and mature EPCs (CD34/CD133/CD309/CD31) were enumerated by flow cytometry. RESULTS: EPCs were reduced in mNPDR (114 +/- 66; P < 0.001) and msNPDR (77 +/- 40; P = 0.042) compared with CO (244 +/- 115). In contrast, EPCs were unchanged in mmPDR (248 +/- 155) compared with CO. Strikingly, EPCs were augmented in hrPDR (389 +/- 124) compared with all other stages. Numbers of undifferentiated progenitor cells (CPCs) did not differ among CO, mmPDR, and hrPDR. Augmentation (3x) of mature EPCs in hrPDR (325 +/- 118; P < 0.001) compared with CO (100 +/- 49) but against all other stages of DR was observed. The percentage of mature EPCs/EPCs was augmented in an ETDRS classification-dependent manner. CONCLUSIONS: In patients with T1DM with DR, EPCs undergo stage-related regulation. In nonproliferative retinopathy, a reduction of EPCs was observed, and in proliferative retinopathy, a dramatic increase of mature EPCs was observed.