Nephrol Dial Transplant (2002) 17: 422-427
© 2002 European Renal Association-European Dialysis and Transplant Association
Convective and diffusive losses of vitamin C during haemodiafiltration session: a contributive factor to oxidative stress in haemodialysis patients
1 Department of Biochemistry, Lapeyronie Hospital, University of Montpellier I, F-34295 Montpellier, France, 2 Department of Nephrology, Lapeyronie Hospital, University of Montpellier I, F-34295 Montpellier, France, 3 Clinical and Laboratory Research Department, Bellco S.p.A., Mirandola, Italy and 4 INSERM U 500, ‘Epidémiologie des maladies chroniques et du vieillissement’, Montpellier, France
Background. Enhanced oxidative stress in haemodialysis (HD) patients may be considered as a risk factor for accelerated atherosclerosis. Reduced antioxidant defences include impairment in enzyme activities and decreased plasma levels of hydrophilic vitamin C (vit C), and cellular levels of lipophilic vitamin E (vit E).
Methods. We investigated plasma levels of vit C in 19 patients undergoing regular haemodiafiltration (HDF) (mean age 62±7 years) and in 1846 healthy elderly subjects (HS) (mean age 69±5 years). The contribution of convection and diffusion was determined using paired filtration dialysis (PFD), a modified HDF technique which physically separates convective from diffusive fluxes. Blood samples were collected before and after the HDF session; in addition at 60 min of HDF, samples were drawn from arterial lines (AL) and venous lines (VL), dialysate (D) and ultrafiltrate (UF). Blood levels of total vit C were determined using an HPLC fluorescence method. Markers of oxidative stress were also assessed in both populations as follows: levels of malondialdehyde (MDA) were determined by fluorometric assay, measurements of advanced oxidation protein products (AOPP) and glutathione peroxidase (GSH-Px) activity were performed by spectrophotometric assay, and plasma vit E content was obtained by an HPLC procedure.
Results. A significant reduction in plasma vit C level was observed in HDF patients when compared with HS (1.6±1.4 µg/ml in HDF vs 6.6±3.7 µg/ml in HS; P<0.01). The HDF session was associated with a dramatic reduction in vit C levels (1.87±1.57 µg/ml before HDF and 0.98±0.68 µg/ml after HDF); at 60 min of HDF, concentrations were as follows: AL=1.35±1.27 µg/ml; VL=0.37±0.31 µg/ml, D=0.40±0.34 µg/ml, UF=1.24±1.18 µg/ml; corresponding to a diffusive flux of 271 µg/min and a convective flux of 126 µg/min. Total loss of vit C could be assessed at 66 mg/session (8–230 mg/session). According to this loss of vit C, presence of an oxidative stress was demonstrated in HD population as shown by a significant increase in MDA (1.66±0.27 µM in HD vs 0.89±0.25 µM in HS; P<0.01) and AOPP (77.5±29.3 µM in HD vs 23.5±13.2 µM in HS; P<0.01) levels, and a decrease in GSH-Px activity (259.2±106.3 U/l in HD vs 661.2±92.2 U/l in HS; P<0.01). No change in plasma vit E between both populations (30.7±9.1 µM in HD vs 35.3±7.34 µM in HS) was observed.
Conclusions. These results suggest that HDF with highly permeable membranes is associated with a significant loss of vit C. Diffusive transport is responsible for two-thirds whereas convective phenomenon accounts for only one-third of this loss.
Keywords: antioxidant; haemodiafiltration; oxidative stress; vitamin C
Correspondence and offprint requests to: Prof. Jean-Paul Cristol, Biochemistry, Lapeyronie University Hospital, 371 Av Doyen G. Giraud, F-34295 Montpellier, France. Email: jp\|[hyphen]\|cristol{at}chu\|[hyphen]\|montpellier.fr
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