FP795DIRECT MEASUREMENT OF REACTIVE OXYGEN SPECIES (ROS) IN CIRCULATING LEUKOCYTES DURING HEMODIALYSIS THERAPY

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Introduction and Aims: Chronic kidney disease (CKD) augments oxidative stress, resulting in increasing mortality of cardiovascular disease (Ikizier TA. Clin Nephrol 2002; 58: 190-197). Hemodialysis (HD) itself may further contribute to progress atherosclerosis by enhancing oxidative stress (Sela S. Kidney Int 2001; 78: S159-S163). Available evidences for oxidative stress come from measurement of byproducts or end-products by reactive oxygen species (ROS). The aim of the present study is to measure directly ROS levels in circulating leukocytes during HD therapy and to examine its usefulness. Methods: Circulating blood cells were collected from HD patients during treatment and from healthy control. Using flow cytometry, the cells were gated for lymphocytes, monocytes, and polymorphonuclear leukocytes (PMN). The cells with increasing levels of ROS demonstrate bright green fluorescence in presence of the ROS detection solution. The serum was collected at the same time with blood cells and used for measurement of circulating F2-isoprostanes. F2-isoprostanes are considered as the best available biomarker of lipid peroxidation and has been investigated in pathogenesis of CKD (Basu S. Free Radic Res 2004; 38: 105-122). We examined correlation between serum F2-isoprostanes levels and ROS production in leukocytes during HD therapy.

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