title: A novel mechanism for hypofibrinolysis in diabetes: the role of complement C3.
abstract: OBJECTIVE
  Impaired fibrin clot lysis is a key abnormality in diabetes and complement C3 is one protein identified in blood clots. This work investigates the mechanistic pathways linking C3 and hypofibrinolysis in diabetes using ex vivo/in vitro studies.
  
  
  METHODS
  Fibrinolysis and C3 plasma levels were determined in type 1 diabetic patients and healthy controls, and the effects of glycaemia investigated. C3 incorporation into fibrin clots and modulation of fibrinolysis were analysed by ELISA, immunoblotting, turbidimetric assays and electron and confocal microscopy.
  
  
  RESULTS
  Clot lysis time was longer in diabetic children than in controls (599 ± 18 and 516 ± 12 s respectively; p < 0.01), C3 levels were higher in diabetic children (0.55 ± 0.02 and 0.43 ± 0.02 g/l respectively; p < 0.01) and both were affected by improving glycaemia. An interaction between C3 and fibrin was confirmed by the presence of lower protein levels in sera compared with corresponding plasma and C3 detection in plasma clots by immunoblot. In a purified system, C3 was associated with thinner fibrin fibres and more prolongation of lysis time of clots made from fibrinogen from diabetic participants compared with controls (244 ± 64 and 92 ± 23 s respectively; p < 0.05). Confocal microscopy showed higher C3 incorporation into diabetic clots compared with controls, and fully formed clot lysis was prolonged by 764 ± 76 and 428 ± 105 s respectively (p < 0.05). Differences in lysis, comparing diabetes and controls, were not related to altered plasmin generation or C3-fibrinogen binding assessed by plasmon resonance.
  
  
  CONCLUSIONS
  C3 incorporation into clots from diabetic fibrinogen is enhanced and adversely affects fibrinolysis. This may be one novel mechanism for compromised clot lysis in diabetes, potentially offering a new therapeutic target.
authors.affiliation: University of Leeds, Leeds, UK.
authors.forename: K
authors.initials: K
authors.lastname: Hess
authors.forename: S H
authors.initials: SH
authors.lastname: Alzahrani
authors.forename: M
authors.initials: M
authors.lastname: Mathai
authors.forename: V
authors.initials: V
authors.lastname: Schroeder
authors.forename: A M
authors.initials: AM
authors.lastname: Carter
authors.forename: G
authors.initials: G
authors.lastname: Howell
authors.forename: T
authors.initials: T
authors.lastname: Koko
authors.forename: M W J
authors.initials: MW
authors.lastname: Strachan
authors.forename: J F
authors.initials: JF
authors.lastname: Price
authors.forename: K A
authors.initials: KA
authors.lastname: Smith
authors.forename: P J
authors.initials: PJ
authors.lastname: Grant
authors.forename: R A
authors.initials: RA
authors.lastname: Ajjan
mesh_terms: D000293:Adolescent; D000328:Adult; D001778:Blood Coagulation Disorders / Q000209:etiology* / Q000378:metabolism; D001780:Blood Coagulation Tests; D016022:Case-Control Studies; D003176:Complement C3 / Q000378:metabolism*; D003922:Diabetes Mellitus, Type 1 / Q000150:complications* / Q000378:metabolism; D005260:Female; D005337:Fibrin / Q000378:metabolism*; D005340:Fibrinogen / Q000378:metabolism*; D005342:Fibrinolysis / Q000502:physiology*; D006801:Humans; D008297:Male
chemical_list: D003176:Complement C3; D005337:Fibrin; D005340:Fibrinogen