ISSN 1607-6729, Doklady Biochemistry and Biophysics, 2018, Vol. 478, pp. 41–43. © Pleiades Publishing, Ltd., 2018.
Original Russian Text © Z.I. Krutetskaya, L.S. Milenina, A.A. Naumova, S.N. Butov, V.G. Antonov, A.D. Nozdrachev, 2018, published in Doklady Akademii Nauk, 2018,
Vol. 478, No. 5, pp. 601–603.
Inhibitors Modulate the Effect of Trifluoperazine
on the Intracellular Ca
Concentration in Macrophages
Z. I. Krutetskaya*, L. S. Milenina, A. A. Naumova, S. N. Butov, V. G. Antonov,
A. D. Nozdrachev
Received June 22, 2017
Abstract—Using Fura-2AM microfluorimetry, it was shown for the first time that phospholipase A
tors 4-bromophenacyl bromide and glucocorticosteroids prednisolone and dexamethasone attenuate Ca
responses induced by neuroleptic trifluoperazine in macrophages. The results suggest the involvement of
and arachidonic acid metabolism cascade in the effect of trifluoperazine on intracellular
concentration in macrophages.
Trifluoperazine (TFP, triftazin) belongs to the first
generation of typical neuroleptics of the phenothi-
azine series, which are widely used in the treatment of
schizophrenia and other mental disorders . The
multifactorial effect of TFP on cellular processes was
established . For example, it was shown  that
phenothiazines (TFP, chlorpromazine) affect the
-dependent activation of macrophages and have
an immunomodulatory effect on rat peritoneal mac-
Earlier , we for the first time showed that, in a
-containing medium, TFP causes a dose-depen-
dent increase in the intracellular Ca
) in rat peritoneal macrophages. However, the
mechanisms underlying the TFP-induced increase in
in macrophages are not fully understood.
The plurality of TFP effects may be due to its
amphiphilic nature, which enables its penetration
through cell membranes. It was found [5, 6] that TFP
interacts mostly with the inner monolayer of the plas-
malemma, in which the anionic phospholipids (pri-
marily phosphoinositides) are located. Due to this
fact, TFP can regulate intracellular processes such as
signaling and intracellular transport. In addition,
experiments on human platelets showed that psycho-
tropic compounds (e.g., TFP) can modulate the acti-
vity of phospholipase A
), the key enzyme of
the arachidonic acid (AA) metabolism cascade [5, 7].
In view of above, it was reasonable to study the pos-
sible involvement of PLA
and arachidonic acid (AA)
metabolic cascade in the effect of TFP on [Ca
macrophages, which was the subject of this communi-
In experiments, we used three PLA
bromophenacyl bromide (4-BPB)  and synthetic
glucocorticoids dexamethasone and prednisolone .
Experiments were performed on cultured residen-
tial peritoneal macrophages of Wistar rats at room
temperature (20–22°C) 1–2 days after the beginning
of cell culturing. The macrophage culturing procedure
and the automated system for measuring [Ca
the basis of Leica DM 4000B fluorescence micro-
scope (Leica Microsystems, Germany) were described
in detail earlier . [Ca
was measured using the
Fura-2AM fluorescent probe (Sigma-Aldrich, United
States). Fluorescence of objects was excited at wave-
lengths of 340 and 380 nm, and emission was recorded
at 510 nm. To prevent photobleaching, measurements
were performed every 20 s irradiating the object for 2 s.
values were calculated using the Gryn-
kiewicz equation . Statistical analysis was per-
formed using Student’s t test. Differences were con-
sidered significant at р ≤ 0.05.
The figure shows the results of typical experiments.
Data are represented as plots showing the changes in
the ratio of Fura-2AM fluorescence intensities at exci-
tation wavelengths 340 and 380 nm (F
time, reflecting the dynamics of changes in [Ca
cells depending on the measurement time .
In the control experiments, we found that the addi-
tion of TFP (4 μg/mL) to macrophages in normal
saline containing Ca
ions caused a rapid increase in
compared to the basal level (92 ± 17 nM), after
which a long-term plateau phase of Ca
observed (Fig. 1a), in which [Ca
was 176 ± 26 nM.
AND MOLECULAR BIOLOGY
St. Petersburg State University, St. Petersburg, 199034 Russia