RESUMO
Production of reactive oxygen species (ROS) by macrophages from blood monocytes of healthy donors (MP(N)) and patients with IHD (MP(IHD)) before, during, and after their incubation with low-density lipoprotein (LDL) isolated from the blood plasma of healthy donors (LDL(N)) and patients with a high cholesterol level (LDL(H)) was estimated by the method of luminol-dependent and stimulated by opsonized zymosan (OZ) or phorbol-12-myristate-13-acetate (PMA) chemiluminescence (CL). Intrinsic luminol-dependent, and zymosan- or PMA-stimulated chemiluminescence of MP(IHD) have exceeded the same types of chemiluminescence of MP(N) by factors of 1.4, 1.8, 2.7, and 1.6, respectively (p < 0.05-0.01). The effect of zymosan on MP(N) and MP(IHD) was stronger than that of TPA by factors of 4.3 and 3.2, respectively, but manifested itself 2.5-3.0 times slower. LDL(N) and LDL(H) incubated with MP(N) during 15-60 min increased ROS production by a factor of 1.4 and 2.5 respectively, but influenced ROS production by MP(IHD) (as estimated by luminol-dependent chemiluminescence). Effects LDL(N) and LDL(H) on MP(IHD) were not detected at all. Repeated increase in zymosan-stimulated CL of MP(N) was also observed after their 15-180 min preincubation with LDL(N) and LDL(H) which followed after taking out LDL, washing MP(N) and adding Hanks' solution with opsonized zymosan. This increase was also stronger after MP(N) incubation with LDL(H) than after MP(N) incubation with LDL(N), and no increase was observed in experiments with MP(IHD). Thus, the results obtained by a chemiluminescent method showed that fresh macrophages from the blood of patients with IHD had higher ROS production than macrophages from healthy donors. LDL(N) and LDL(H) could exhibit primary and secondary (after preincubation) stimulating effect on CL in MP(N); but had no effect on MP(IHD). An analysis of macrophage chemiluminescence is a sensitive test for evaluation the degree of macrophage's stimulation and it may be effectively used for the dynamic control for treatment effectiveness in clinics.
