title: Coagulation activation is associated with nicotinamide adenine dinucleotide phosphate oxidase-dependent reactive oxygen species generation in hemodialysis patients.
abstract: OBJECTIVE
  This study investigated on (i) the role of gp91(phox)/NOX2 in reactive oxygen species (ROS) generation in hemodialysis (HD) patients, and (ii) the link between clotting activation and ROS production in this setting.
  
  
  RESULTS
  The study was performed on peripheral blood mononuclear cells (PBMCs) isolated from HD patients randomized to polysulphon/polyamide (S-group, n=30) or ethylene-vinyl-alcohol (EVAL) membrane (E-group, n=30) treatment and from healthy subjects (control group, n=15). ROS generation was increased in PBMCs of HD patients compared with healthy subjects. S-group showed higher levels of intracellular ROS generation than control, whereas E-group did not. In addition, S-group displayed an increase in nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity compared with E-group and healthy subjects. A further increase in NADPH activity shortly after HD treatment was observed only in S-group. The plasma levels of the prothrombin fragment F1+2, a marker of in vivo clotting activation, were significantly higher in S-group than in E-group. Moreover, a heightened thrombin generation was recorded in the plasma of S-group. Intracellular ROS production correlated with NADPH oxidase activity and coagulation priming in HD patients. The in vitro validation study demonstrated that incubation of PBMCs with activated FX induced a significant increase in intracellular ROS production, superoxide generation, and gp91(phox)/NOX2 expression.
  
  
  METHODS
  The pivotal role of NADPH oxidase in the upregulation of ROS in HD patients makes this enzyme a potential target for therapeutic intervention in the treatment of HD-related oxidative stress.
  
  
  CONCLUSIONS
  The EVAL membrane, by reducing clotting activation, inhibits gp91(phox)/NOX2-related ROS production in HD patients.
authors.affiliation: Renal, Dialysis and Transplantation Unit, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Marica
authors.initials: M
authors.lastname: Cariello
authors.forename: Simona
authors.initials: S
authors.lastname: Simone
authors.forename: Antonia
authors.initials: A
authors.lastname: Loverre
authors.forename: Margherita
authors.initials: M
authors.lastname: Gigante
authors.forename: Francesca
authors.initials: F
authors.lastname: Incampo
authors.forename: Stefania
authors.initials: S
authors.lastname: Pietanza
authors.forename: Mario
authors.initials: M
authors.lastname: Colucci
authors.forename: Francesco P
authors.initials: FP
authors.lastname: Schena
authors.forename: Loreto
authors.initials: L
authors.lastname: Gesualdo
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Grandaliano
authors.forename: Giovanni
authors.initials: G
authors.lastname: Pertosa
mesh_terms: D000328:Adult; D005260:Female; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008297:Male; D008567:Membranes, Artificial; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009757:Nylons / Q000737:chemistry / Q000494:pharmacology; D011145:Polyvinyls / Q000737:chemistry / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D006435:Renal Dialysis; D013450:Sulfones / Q000737:chemistry / Q000494:pharmacology; D013499:Surface Properties
chemical_list: D008567:Membranes, Artificial; D009757:Nylons; D011145:Polyvinyls; D017382:Reactive Oxygen Species; D013450:Sulfones; C020320:ethylene-vinyl alcohol copolymer; D019255:NADPH Oxidases