Effects of Hemodialysis, Dialyser Type and Iron Infusion on Oxidative Stress in Uremic Patients

Abstract

Uremic patients undergoing hemodialysis (HD) are considered to face an elevated risk for atherosclerosis and cancer. This has been attributed in part to an increased oxidative stress. In this pilot study, oxidative cell damage in blood of HD-patients was compared to those of controls: total DNA damage (basic and specific oxidative DNA damage), modulation of glutathione levels (total and oxidized glutathione) and of lipid peroxidation were monitored via the Comet assay (with and without FPG), a kinetic photometric assay and HPLC quantification of plasma malondialdehyde (MDA), respectively. In some samples, leukocytes were analysed for malondialdehyde–deoxyguanosine-adducts (M₁dG) with an immunoslot blot technique.

HD-patients (n=21) showed a significant increase of total DNA damage (p<10^-12), compared to controls (n=12). In a subset of patients and controls, GSSG levels and M₁dG, however, only increased slightly, while tGSH and MDA levels did not differ. The influence of different low flux HD-membranes was tested in a pilot study with nine patients consecutively dialysed on three membrane types for four weeks each. In addition to the individual disposition of the patient, the dialyser membrane had a significant impact on oxidative stress. Total DNA damage was found to be almost identical for polysulfone and vitamin E coated cellulosic membranes, whereas a slight, but significant increase was observed with cellulose-diacetate (p<0.001). In patients receiving iron infusion during HD, MDA-formation (n=11) and total DNA damage (n=10) were additionally increased (p<0.005).

Our results show an increased oxidative damage in HD-patients, compared to healthy volunteers. Significant influences were found for the dialyser membrane type and iron infusion.

• Oxidative stress
• Hemodialysis
• Dialyser membrane
• Iron infusion
• Comet assay
• AGE, advanced glycation end products
• ANOVA, analysis of variance
• BHT, butylated hydroxytoluene
• DTNB 5, 5′-dithiobis(2-nitrobenzoic acid)
• ESRD, end-stage renal disease
• FPG, formamidopyrimidine-DNA N-glycosylase
• GSH, reduced glutathione
• GSSG, oxidised glutathione
• HD, hemodialysis
• M1dG, malondialdehyde–deoxyguanosine-adduct
• MDA, malondialdehyde
• ROS, reactive oxygen species
• SSA, 5-sulfosalicylic acid
• tGSH, total glutathione
• TI%, tail intensity in percent
• TNB, 5-thio-2-nitrobenzoate