1. Orekhovich Institute of Biomedical Chemistry, Russian Academy of Medical Sciences 2. Orekhovich Institute of Biomedical Chemistry, Russian Academy of Medical Sciences Faculty of Basic Medicine, Moscow State University
Abstract: Production of reactive oxygen species (ROS) by macrophages from blood monocytes of healthy donors (MPN) and patients with IHD (MPIHD) before, during, and after their incubation with low-density lipoprotein (LDL) isolated from the blood plasma of healthy donors (LDLN) and patients with a high cholesterol level (LDLH) was estimated by the method of luminol-dependent and stimulated by opsonized zymosan (OZ) or phorbol-12-myristate-13-acetate (PMA) сhemiluminescence (CL). Intrinsic luminol-dependent, and zymosan - or PMA-stimulated chemiluminescence of MPIHD have exceeded the same types of chemiluminescence of MPN by factors of 1.4, 1.8, 2.7, and 1.6, respectively (p<0.05-0.01). The effect of zymosan on MPN and MPIHD was stronger than that of TPA by factors of 4.3 and 3.2, respectively, but manifested itself 2.5-3.0 times slower. LDLN and LDLH incubated with MPN during 15-60 min increased ROS production by a factor of 1.4 and 2.5 respectively, but influenced ROS production by MPIHD (as estimated by luminol-dependent chemiluminescence). Effects LDLN and LDLH on MPIHD were not detected at all. Repeated increase in zymosan-stimulated CL of MPN was also observed after their 15-180 min preincubation with LDLN and LDLH which followed after taking out LDL, washing MPN and adding Hanks' solution with opsonized zymosan. This increase was also stronger after MPN incubation with LDLH than after MPN incubation with LDLN, and no increase was observed in experiments with MPIHD. 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. LDLN and LDLH could exhibit primary and secondary (after preincubation) stimulating effect on CL in MPN; but had no effect on MPIHD. 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.
Reference: Bilenko M.V., Vladimirov Yu. A., Pavlova S. A., Nguyen Thi Thu Thuy, Yen Tran Thi Hai, Production of reactive oxygen species by monocyte-derived macrophages from the blood of healthy donors and patients with IHD, Biomeditsinskaya khimiya, 2008, vol:
5. Bilenko M.V. (2001) Ischemia and reperfusion of various organs. Injury mechanisms, methods of prevention and treatment. (S. Boriotti and D. Denniss eds.), Nova Science Publishers, Inc. Huntington, New York.