Coagulation activation is associated with nicotinamide adenine dinucleotide phosphate oxidase-dependent reactive oxygen species generation in hemodialysis patients

Antioxid Redox Signal. 2012 Mar 1;16(5):428-39. doi: 10.1089/ars.2011.4062. Epub 2011 Dec 22.


Aims: 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.

Innovation: 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.

Conclusion: The EVAL membrane, by reducing clotting activation, inhibits gp91(phox)/NOX2-related ROS production in HD patients.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Female
  • Humans
  • Leukocytes, Mononuclear / metabolism
  • Male
  • Membranes, Artificial
  • NADPH Oxidases / antagonists & inhibitors
  • NADPH Oxidases / metabolism*
  • Nylons / chemistry
  • Nylons / pharmacology
  • Polyvinyls / chemistry
  • Polyvinyls / pharmacology
  • Reactive Oxygen Species / metabolism*
  • Renal Dialysis*
  • Sulfones / chemistry
  • Sulfones / pharmacology
  • Surface Properties


  • Membranes, Artificial
  • Nylons
  • Polyvinyls
  • Reactive Oxygen Species
  • Sulfones
  • ethylene-vinyl alcohol copolymer
  • NADPH Oxidases