EFFECT OF THE 1,4-NAPHTHOQUINONE DERIVATIVE LES6400 ON THE ACTIVITY OF ENZYMES OF THE ARGINASE-NO-SYNTHASE SYSTEM OF SPERMATOZOA
DOI:
https://doi.org/10.11603/1811-2471.2025.v.i2.15276Keywords:
1-(furan-2-yl)-9-hydroxy-3,11-dihydro-2H-benzo[6,7]thiochromeno[2,3-d]thiazole-2,5,10-trione11-(furan-2-yl)-9-hydroxy-3,11-dihydro-2H-benzo[6,7]thiochromeno[2,3-d]thiazole-2,5,10-trione (Les6400), spermatozoa, arginase, NO synthase isoformsAbstract
SUMMARY. In the course of research on 11-(furan-2-yl)-9-hydroxy-3,11-dihydro-2H-benzo[6,7]thiochromeno[2,3-d]thiazole-2,5,10-trione (Les6400), probable molecular biological pathways for the implementation of antitumor action have been outlined. However, there are few works devoted to the detailed establishment of the mechanisms of implementation of the biological activity of this class of compounds.
The aim – to study the effect of 11-(furan-2-yl)-9-hydroxy-3,11-dihydro-2H-benzo[6,7]thiochromeno[2,3-d]thiazole-2,5,10-trione (Les6400) on the activity of enzymes of the arginase-NO synthase system of cells (spermcells).
Material and Methods. The study included 29 semen samples from men with normozoospermia. Determination of arginase activity of spermatozoa permeabilized with saponin was carried out by urea formation. The activity of individual NO synthase isoforms was determined by the concentration of citrulline by a highly specific method using a color reaction with antipyrine.
Results. It was shown that at concentrations of 10-6–10-4 M Les6400 significantly increases arginase activity in spermatozoa. However, its most pronounced effect was observed at 10-5 M concentration. The activity significantly increased by 1.2 times. When Les6400 was applied to cNOS at concentrations of 10-4 M, its activity significantly decreased by 1.21 times. The iNOS activity of spermatozoa of fertile men was quantified being less than cNOS activity and was calculated as 0.28±0.09 pmol citrulline/min·mg protein. When studying the effect of Les6400 on the iNOS activity of spermatozoa of practically healthy men, we did not observe its activation, and the inhibitory effect could not be determined due to the low activity of the enzyme. Preincubation of spermatozoa with 0.2 mM H2O2 led to an increase in iNOS activity by 21.2 times. Against the background of iNOS activation by hydrogen peroxide, Les6400 at concentrations of 10-6–10-4 M leads to inhibition of enzyme activity by 1.3–1.5 times.
Conclusions. It was found that the compound Les6400 leads to an increase in arginase activity and inhibition of both isoforms of NO synthase. Since arginase and NO synthase isoforms compete under physiological conditions in the cell for one substrate – L-arginine, it is likely that an increase in arginase activity leads to a decrease in NO synthase activity.
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