Acute effect of H.E.L.P. treatment on radical scavenging enzyme activities, total glutathione concentrations in granulocytes, and selenium in plasma.

Abstract

It has been suggested that granulocytes are activated on artificial surfaces such as dialyzer membranes or by plasma separation procedures resulting in the generation of free radicals. We reported recently that free radical scavenging enzyme (FRSE) activities of red blood cells obtained from patients undergoing hemodialysis and LDL-apheresis (LA) do not reflect an acute oxidative stress. However, because mature red cells are free of DNA and RNA, enzymes cannot be regulated on the gene level. In contrast, granulocytes are nucleated cells in which genes can be regulated, e. g. by redox sensitive transcription factors activated by extracellular oxidative stress. Therefore, granulocyte FRSE may better reflect acute oxidative stress caused by extracorporeal treatment. Hyperlipidemic patients (n = 18) with coronary heart disease (CHD) were treated with the Heparin-induced-Extracorporeal-LDL-Precipitation (H.E.L.P.) system. Glutathione peroxidase (GSH-Px), glutathione reductase (GSSG-R), superoxide dismutase (SOD) activities, and total glutathione were determined in granulocytes before and immediately after a single LA treatment. Selenium (Se) concentrations were assessed in plasma. As a result of the H.E.L.P. treatment GSSG-R activity was significantly induced (+ 20%) and the GSH concentration increased (+ 41%) in granulocytes. GSH-Px activity in granulocytes (- 19%) and Se in plasma (- 27%) were significantly reduced whereas SOD activity in granulocytes was not affected by the H.E.L.P. procedure. These results show that the defence against oxygen radicals in granulocytes is affected but not severely compromised in patients undergoing regular H.E.L.P-LDL-apheresis treatment, which points to the safety of this system with respect to oxidative stress.