Increased intracellular reactive oxygen species in patients with end-stage renal failure: Effect of hemodialysis

Abstract

Reactive oxygen species (ROS) have been implicated in various forms of cellular injury. ROS may cause cell damage and are involved in the pathophysiology of several diseases, including atherosclerosis and chronic inflammation. Disturbances of intracellular ROS levels were investigated in 28 patients with end-stage renal failure. The intracellular ROS levels were measured in lymphocytes before and after hemodialysis using biocompatible membranes and were compared with those from 11 patients with end-stage renal failure, not yet on renal replacement therapy, and 27 healthy control subjects. ROS levels were measured spectrophotometrically using the intracellular dye dichlorofluorescin diacetate. The spontaneous production of ROS was significantly higher in lymphocytes from patients with end-stage renal failure compared with healthy control subjects (P < 0.01). The addition of 100 nmol/L phorbol-myristate-acetate (PMA) produced a significant increase of ROS, both in lymphocytes from patients with end-stage renal failure and healthy control subjects. The PMA-induced ROS increase was significantly higher in lymphocytes from patients with end-stage renal failure compared with healthy control subjects (P < 0.01). In patients with end-stage renal failure, not yet on renal replacement therapy, the PMA-induced ROS was also significantly higher compared with healthy control subjects. The PMA-induced ROS increases were significantly inhibited by catalase, but not by superoxide dismutase or the superoxide dismutase mimetic, tempol. PMA-induced ROS was significantly reduced by tyrphostin A51 in lymphocytes from patients with end-stage renal failure and from healthy control subjects (each P < 0.01), indicating the involvement of a tyrosine kinase-dependent pathway. In patients with end-stage renal failure, the spontaneous and the PMA-induced production of ROS was not significantly different before and after hemodialysis. Regular hemodialysis sessions using biocompatible membranes have no effect on the elevated intracellular ROS in patients with end-stage renal failure.