Аrginase/NO-synthase system сharacteristics in blood lymphocytes under effect of fluoroquinolones

Abstract

Antibiotics of the fluoroquinolone series are highly effective synthetic drugs of a wide range of antimicrobial activity. They have a specific mechanism of action on bacterial cells to inhibit the DNA-gyrase enzyme of mostly gram-negative and the topoisomerase IV of gram-positive bacterial cells, which leads to a decrease in the activity of enzymes, disruption of DNA and RNA biosyntheses, and the impossibility of chromosome superspilarization, as a result of which its division is broken and the cell dies. Fluoroquinolones also have an immunomodulatory effect, which is very important in terms of the pathogenesis of many infectious and inflammatory diseases. We assume that fluoroquinolones also act on cells of the body of patients, in particular on such regulatory mechanisms as the arginase-NO-synthase system. In this regard, peripheral blood lymphocytes can be a convenient and adequate model for studying the mechanism of the effect of fluoroquinolones. It is shown that under the influence of various generations of fluoroquinolones, arginase activity increases, depending on the dose, in the following sequence: control → ciprofloxacin → levofloxacin → moxifloxacin. The highest activity is observed under the effects of moxifloxacin, belonging to generation IV. The increase in arginase activity in blood lymphocytes under the influence of fluoroquinolones occurs due to the growth of turnover number of the enzymes (Vmax increases), although the affinity of enzyme to the substrate decreases (KL-arg increases). At the same time, all fluoroquinolones reduce the activity of the constitutive isoforms of NO-synthase, depending on the dose. It has been established that a slight activity of iNOS of blood lymphocytes in practically healthy women was detected, almost on the verge of error. When studying the influence of fluoroquinolones on the activation of iNOS lymphocytes isolated from the blood of practically healthy women, we did not observe its activity, and the inhibitive effect could not be determined due to its low activity. Oxidative stress was used to induce iNOS activity in blood lymphocytes, with H2O2 lymphocytes preincubation. The preincubation of lymphocytes with 0.2 mM H2O2 leads to increase of iNOS activity by 31.30 times. By activation of iNOS with hydrogen peroxide, 10–5 M concentration of ciprofloxin leads to inhibition of enzyme activity by 1.22 times, levofloxacin by 1.45 and moxifloxacin by 2.34 times. The obtained kinetic parameters suggest that in the blood lymphocytes under the influence of fluoroquinolones, the synthesis of NO with the participation of cNOS is inhibited, and the hyperproduction of NO is inhibited by the activation of iNOS, which is characteristic for pathological conditions.