Molecular mechanisms involved in the resistance of fibrin to clot lysis by plasmin in subjects with type 2 diabetes mellitus.
OBJECTIVE
The aim of this study was to determine the influence of type 2 diabetes on fibrinolysis by assessing interactions between the regulatory components of fibrinolysis and the fibrin clot, using fibrinogen purified from 150 patients with type 2 diabetes and 50 matched controls.


METHODS
Clot lysis rates were determined by confocal microscopy. Plasmin generation was measured using a plasmin-specific chromogenic substrate. Surface plasmon resonance was used to determine the binding interactions between fibrin, tissue-type plasminogen activator (t-PA) and Glu-plasminogen; cross-linkage of plasmin inhibitor to fibrin by factor XIII was determined using a microtitre plate assay.


RESULTS
Lysis of diabetic clots was significantly slower than that of controls (1.35 vs 2.92 microm/min, p<0.0001) and plasmin generation was significantly reduced. The equilibrium binding affinity between both t-PA and Glu-plasminogen and fibrin was reduced in diabetic subjects: t-PA, K (D)=0.91+/-0.3 micromol/l (control subjects), 1.21+/-0.5 micromol/l (diabetic subjects), p=0.001; Glu-plasminogen, K (D)=97+/-19 nmol/l (control subjects), 156+/-66 nmol/l (diabetic subjects), p=0.001. Cross-linkage of plasmin inhibitor to fibrin by factor XIII was enhanced in diabetic subjects, with the extent of in vitro cross-linkage correlating with in vivo glycaemic control (HbA(1c)) (r=0.59, p=0.001).


CONCLUSIONS
These results indicate that impairment of the fibrinolytic process in diabetic patients is mediated via a number of different mechanisms; these may be a consequence of post-translational modifications to fibrinogen molecules, resulting from their exposure to the abnormal metabolic milieu associated with diabetes.
Dunn
E J
EJ
Academic Unit of Molecular Vascular Medicine, Leeds Institute for Genetics, Health and Therapeutics, University of Leeds, Clarendon Way, Leeds LS1 3EX, UK.
Philippou
H
H
Ariëns
R A S
RA
Grant
P J
PJ
D000368:Aged; D001786:Blood Glucose / Q000378:metabolism; D015992:Body Mass Index; D003924:Diabetes Mellitus, Type 2 / Q000097:blood*; D004351:Drug Resistance; D004789:Enzyme Activation; D005176:Factor XIII / Q000302:isolation & purification; D005260:Female; D005337:Fibrin / Q000494:pharmacology*; D005340:Fibrinogen / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D005341:Fibrinolysin / Q000494:pharmacology*; D006442:Glycated Hemoglobin / Q000378:metabolism; D006461:Hemolysis / Q000187:drug effects*; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D012016:Reference Values; D020349:Surface Plasmon Resonance
D001786:Blood Glucose; D005176:Factor XIII; D005337:Fibrin; D005340:Fibrinogen; D005341:Fibrinolysin; D006442:Glycated Hemoglobin