Abstract
Luminol-enhanced chemiluminescence (LCL) is a widely used method for quantitative evaluation of reactive oxygen species (ROS) generated by stimulated leukocytes. The chemiluminescent kinetics registered (Total LCL) is an expression of simultaneously running processes of extra- and intracellular radical generation. In literature, model chemiluminescent systems, designed to separate extra- from the intracellular light, have been proposed. These systems, however, change cellular activity. The aim of the present study was to propose an approach for evaluating the intracellular ROS generation by stimulated leukocytes in whole blood. Heparin anti-coagulated blood from healthy volunteers was used. The chemiluminescent kinetics reflecting leukocyte activity to produce ROS were registered by a computerized chemiluminometer. To distinguish extra - from the intracellularly generated light by the cells, some model chemiluminescent systems were used. It was found that the intracellular light generated by stimulated leukocytes was a substantial part of Total LCL. The proportion between the extra- and intracellular light within the Total response depended on the concentration of the particulate stimulant. At a low zymosan concentration (0.25 mg/ml) Total LCL response was found to be intracellular. On the basis of the results obtained we conclude that Total LCL at a low stimulant concentration is of an intracellular origin, which could be used to study the effect of different compounds on the intracellular oxidative leukocyte activity. In the course of some pathological processes accompanied by suppurative foci formation, bacteriemia, ect., the proportion between the number of bacteria and leukocytes considerably changes. A change in the stimulant concentration allows to model these pathological states and to evaluate the extra- and intracellular radical generation by stimulated leukocytes. Scripta Scientifica Medica 2007;39(2):111-114
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