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BULLETIN OF EXPERIMENTAL
BIOLOGY AND MEDICINE
September, 2008Volume 146, Number 3
A translation of
Byulleten’ Eksperimental’noi Biologii i Meditsiny
and
Kletochnye Technologii v Biologii i Meditsine
CONTENTS Engl./Russ.
Translated from
Byulleten’ Eksperimental’noi Biologii i Meditsiny
(Bulletin of Experimental Biology and Medicine)
PHYSIOLOGY
Anticoagulant Activity and Platelet Aggregation in Blood Plasma
from Rats after Intranasal Administration of Adenosine Triphosphate
and Its Complex with Heparin
W. E. Pastorova, L. A. Lyapina, T. Y. Obergan, and T. A. Shubina....................................................
273 244
Effect of Muramyl Dipeptides on Postsynaptic GABA, NMDA, and AMPA
Receptors and Presynaptic NMDA Receptors in Rat Brain
V. V. Grigoriev, L. N. Petrova, A. V. Gabreliyan, and T. A. Ivanova ...................................................
276 247
GENERAL PATHOLOGY AND PATHOPHYSIOLOGY
Cytomorphological Study of the Development of Fibrotic Complications
in Chronic SiO2 Granulomatosis in the Liver during Radon Treatment
M. S. Novikova, O. V. Potapova, and V. A. Shkurupy ...........................................................................
279 250
Free Radical Lipid Peroxidation during Amiodarone Therapy
for Postinfarction Cardiosclerosis
T. Yu. Rebrova and S. A. Afanasyev ........................................................................................................
283 254
Blood Lymphocyte Proliferation Reaction in Autoimmune Thyropathies
O. I. Urazova, E . B. Kravets, Yu. V. Nedosekova, A. V. Rogaleva,
V. N. Kuznecova, and O. A. Rumpel ........................................................................................................
286 257
Prevention of Experimental Stroke by Hypercapnic-Hypoxic Preconditioning
N. N. Yakushev, A. G. Bespalov, and V. P. Kulikov ..............................................................................
289 261
Lipid Peroxidation in Various Organs and Tissues of Albino Rats
with Cadmium Intoxication in Winter and Summer
S. V. Kotelnikova, N. G. Sokolova, and A. V. Kotelnikov ......................................................................
291 264
Use of Terahertz Electromagnetic Radiation for Correction of Blood
Rheology Parameters in Patients with Unstable Angina under Conditions
of Treatment with Isoket, an NO Donor
V. F. Kirichuk, E. V. Andronov, N. V. Mamontova,
V. D. Tupicin, and A. V. Mayborodin ........................................................................................................
293 266
Erythropoiesis and Iron Metabolism Biorhythms in Children with Chronic
Pyelonephritis
E. N. Barkova, K. A. Lebedeva, and E. P. Ashikhmina .........................................................................
297 271
BIOPHYSICS AND BIOCHEMISTRY
DNA Damage in Human Mononuclear Cells Induced by Bacterial Endotoxin
I. L. Glukhov, N. P. Sirota, and E. A. Kuznetsova ..................................................................................
301 275
Changes in Activity of Neutral Proteases in Tissues of Ground Squirrels
in the Dynamics of Hibernation
E. Z. Emirbekov, P. M. Nurmagomedova, and M. M. Abasova ............................................................
304 278
PHARMACOLOGY AND TOXICOLOGY
Methodological Aspects of Studies of Chemical Mutagenesis Modification
A. D. Durnev..................................................................................................................................................
307 281
Pharmacokinetics of Coenzyme Q10
E. I. Kalenikova, E. A. Gorodetskaya, and O. S. Medvedev .................................................................
313 288
Expression of mRNA for Corticotropin-Releasing Hormone and Vasopressin
in the Hypothalamus and Amygdala of Rats after Administration
of Narcogenic
P. D. Shabanov and A. A. Lebedev .........................................................................................................
317 292
Effect of Nadcin on Energy Supply System and Apoptosis
in Ischemia-Reperfusion Injury to the Myocardium
G. V. Sukoyan and I. K. Kavadze ..............................................................................................................
321 297
Effect of Desmopressin on Erythrocyte Aggregation
N. P. Zdumaeva and V. N. Levin ..............................................................................................................
325 301
IMMUNOLOGY AND MICROBIOLOGY
Inhibition of Thrombin and Factor Xa by Fucus Evanescens Fucoidan
and Its Modified Analogs
E. S. Lapikova, N. N. Drozd, A. S. Tolstenkov, V. A. Makarov,
T. N. Zvyagintseva, N. M. Shevchenko, I. U. Bakunina,
N. N. Besednova, and T. A. Kuznetsova ..................................................................................................
328 304
Noopept Stimulates the Expression of NGF and BDNF in Rat
Hippocampus
R. U. Ostrovskaya, T. A. Gudasheva, A. P. Zaplina, Ju. V. Vahitova,
M. H. Salimgareeva, R. S. Jamidanov, and S. B. Seredenin ...............................................................
334 310
Study of Heterogeneity of
Coxiella burnettii
Strains by Analysis of groEL
Gene Restriction Fragment Length Polymorphism
O. A. Freylikhman, Y. A. Panfyorova, and N. K. Tokarevich .................................................................
338 314
VIROLOGY
Studying the Pathogenicity of Avian Influenza Virus Subtype H5N1
Strains from the Russian Federation Using Mouse Model
A. M. Shestopalov, K. A. Sharshov, A. V. Zaykovskaya, Yu. N. Rassadkin,
I. G. Drozdov, V. A. Shkurupy, O. V. Potapova, and N. G. Luzgina....................................................
341 317
EXPERIMENTAL GENETICS
Aneuploidy of Stem Cells Isolated from Human Adipose Tissue
N. P. Bochkov, V. A. Nikitina, O. A. Buyanovskaya, E. S. Voronina,
D. V. Goldstein, N. P. Kuleshov, A. A. Rzhaninova, and I. N. Chaushev ..........................................
344 320
CONTENTS
(continued)
Engl./Russ.
EXPERIMENTAL METHODS FOR CLINICAL PRACTICE
Contribution of Leptin to the Formation of Neuroleptic Obesity in Patients
with Schizophrenia during Antipsychotic Therapy
L. N. Gorobets ...............................................................................................................................................
348 324
ATHLETIC MEDICINE
Effect of HIF1A Gene Polymorphism on Human Muscle Performance
I. I. Ahmetov, A. M. Hakimullina, E. V. Lyubaeva,
O. L. Vinogradova, and V. A. Rogozkin ...................................................................................................
351 327
Free Testosterone as Marker of Adaptation to Medium-Intensive Exercise
M. U. Shkurnikov, A. E. Donnikov, E. B. Akimov, D. A. Sakharov,
and A. G. Tonevitsky ....................................................................................................................................
354 330
EXPERIMENTAL BIOLOGY
Effect of Epiphysectomy on the Circadian Dynamics of Antiradical Activity
of Chalone-Antichalone System of Albino Rat Liver
S. M. Slesarev, V. I. Arav, A. I. Antohin, and A. N. Pashkov ...............................................................
358 333
MORPHOLOGY AND PATHOMORPHOLOGY
Morphological Changes in Intervertabral Disk Tissues in a Static
Asymmetrical Compression Model of Degenerative Dystrophic
Diseases of Intervertabral Disks
A. V. Volkov, G. B. Bol’shakova, and D. V. Goldstein ...........................................................................
361 336
Ultrastructural Signs of Cyclophosphamide-Induced Damage
to Cardiomyocytes
E. L. Lushnikova, L. M. Nepomnyashchikh, E. A. Sviridov,
and M. G. Klinnikova ...................................................................................................................................
366 341
Morphogenesis of the Dorsomedial Nucleus of the Amygdaloid Complex
in Early Juvenile Period in Rats
A. V. Akhmadeev and L. B. Kalimullina ...................................................................................................
372 347
Role of Nitric Oxide in the Regulation of Activity of Proteinase Inhibitors
α1-Antitrypsin and α2-Macroglobulin by Capsaicin-Sensitive Nerves
V. K. Spiridonov and Z. S. Tolochko.........................................................................................................
375 350
METHODS
Effects of Long-Term Exposure of Nerve Cells to Autoblood in
In Vitro
Model of Hemorrhagic Stroke
A. A. Mokrushin, A. X. Khama-Murad, and L. I. Pavlinova ...................................................................
379 355
A Method for Extirpation of the Pineal Gand in Albino Rats
V. I. Arav, S. M. Slesarev, and E. V. Slesareva......................................................................................
382 358
(continued)
CONTENTS
Engl./Russ.
Bulletin of Experimental Biology and Medicine
is abstracted or indexed in Chemical Abstracts Service, EMBASE,
Index Medicus/MEDLINE, ISI Alerting Services, Medical Documentation Service, Science Citation Index, Science
Citation Index Expanded, SCOPUS.
273
0007-4888/08/14630273 © 2008 Springer Science+Business Media, Inc.
PHYSIOLOGY
Anticoagulant Activity and Platelet Aggregation in Blood
Plasma from Rats after Intranasal Administration
of Adenosine Triphosphate and Its Complex with Heparin
W. E. Pastorova, L. A. Lyapina, T. Y. Obergan, and T. A. Shubina
Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 146, No. 9, pp. 244-246, September, 2008
Original article submitted November 19, 2007
Fourfold intranasal administration of ATP was followed by an increase in anticoagulant
activity and decrease in platelet aggregation in blood plasma of healthy rats and, parti-
cularly, of animals with suppressed function of the anticoagulant system. These changes
decrease the risk of the prethrombotic state. Complex of ATP and high-molecular-weight
heparin decreased platelet aggregation only in the blood from healthy animals, but
increased anticoagulant potential of plasma hemostasis in prethrombotic animals.
Key Words: adenosine triphosphate-heparin complex; anticoagulant activity; platelet
aggregation
Department of Human and Animal Physiology, B. A. Kudryashov
Laboratory of Protective Systems of blood, Biological Faculty, M. V.
Lomonosov Moscow State University, Russia.
Address for corre
spondence:
lyapina@mail.ru. L. A. Lyapina
ATP is a natural high-molecular-weight ligand for-
med during glycolytic degradation of carbohydrates
[9]. Anticoagulant effect of ATP were demonstra-
ted in vitro and in vivo by computer modeling and
in experimental coagulation studies [5,6,8]. An-
other blood anticoagulant, heparin, is present in the
vascular endothelium and blood, is highly reactive
to the formation of complexes with various sub-
stances [10], including ATP [5]. Our previous stu-
dies showed that a complex of ATP and heparin
(ATP-H) demonstrates strong anticoagulant, fibrin-
depolymerizing, and fibrinolytic effects and redu-
ces ADP-induced platelet aggregation in vitro and
after intramuscular injection to healthy animals
[5,7]. Since parenteral treatment with these prepara-
tions causes side effects, they are often admini-
stered intranasally. After intranasal administration,
this drug enters the blood flow, reaches the Jacob-
son’s organ, and contributes to transduction of ner-
ve impulses to brain receptors [2]. The anticoa-
gulant system is of the neurohumoral nature [4].
Aging of the organism is accompanied by a de-
crease in anticoagulant function [1]. It remains un-
clear whether intranasal administration of ATP and
ATP-H can correct plasma and platelet hemostasis
in animals of different age.
This work was designed to study anticoagulant
activity of the plasma and platelet hemostasis in
animals of different age after intranasal administra-
tion of ATP and ATP-H.
MATERIALS AND METHODS
Experiments were performed with 1% ATP sodium
salt. ATP-H was obtained as described elsewhere
[5]. Male laboratory rats were in vivo divided into
3 groups. Group 1 consisted of healthy adult ani-
mals aging 4-4.5 months and weighing 190-210 g.
Bulletin of Experimental Biology and Medicine, Vol. 146, No. 3, September, 2008
274
Groups 2 and 3 included rats aging 8-8.5 (350-380
g) and 10 months (400-430 g), respectively.
The test preparations were administered intra-
nasally at 24-h intervals (4-fold treatment). Each
group of rats was divided into the following sub-
groups: subgroup A, administration of NaCl (control);
subgroup B, administration of ATP; and subgroup
C, administration of ATP-H. The blood was taken
from the jugular vein with a syringe with 3.8% so-
dium citrate (blood/anticoagulant ratio 9:1). Blood
sampling was performed 1-1.25 h after the last treat-
ment with the test preparations. The blood was
centrifuged to study platelet aggregation (1000g,
5.5 min) and blood coagulation (3000g, 12 min).
Blood anticoagulant activity was estimated from
activated partial thromboplastin time (APTT) [3].
Platelet aggregation was evaluated in platelet-rich
plasma after addition of the aggregation-inducing
agent (2-5 µM ADP) by measuring optical density
of plasma samples on an aggregometer (M. V. Lo-
monosov Moscow State University). Platelet aggrega-
tion was expressed as an index of the light scattering
increment (ISL) and in percent of the control value.
The results were analyzed by Student’s t test.
RESULTS
Table 1 shows parameters of platelet aggregation
and anticoagulant activity of blood plasma in rats
of various experimental groups after 4-fold intra-
nasal administration of ATP, ATP-H, and NaCl.
Intranasal administration of ATP (0.96 mg/kg,
0.02 ml) to healthy adult animals was followed by
a slight increase in anticoagulant activity (by 10%)
and decrease in platelet aggregation (by 13%) com-
pared to the control. Similar changes were previ-
ously revealed after intramuscular injection of ATP
to healthy rats [8]. Intranasal administration of
ATP-H (1 mg/kg) induced a significant decrease in
platelet aggregation (by 48%) and increase in anti-
coagulant activity of blood plasma (by 1.14 times)
in animals of this group (Table 1).
Intranasal administration of ATP to 8-8.5-month-
old rats with partial suppression of the anticoagulant
system (ACS) was followed by a significant de-
crease in platelet aggregation (by 49% below the
control, Table 1) [1,4]. Administration of ATP-H
under these conditions had no effect on platelet
aggregation (in contrast to ATP). Anticoagulant
activity of blood plasma increased by 1.16 times
after treatment with ATP and ATP-H.
Intranasal administration of ATP and ATP-H
had a similar effect on platelet aggregation in group
3 animals with severe dysfunction of ACS. The
prethrombotic state in these rats manifested in a de-
crease in anticoagulant and fibrinolytic activity of
the blood and increase in fibrinogen concentration
and platelet aggregation [1]. ATP administration to
these rats was followed by a significant decrease in
platelet aggregation (by 35% compared to the con-
trol). However, platelet aggregation remained un-
changed after administration of ATP-H. Anticoagu-
lant activity increased by 1.3 and 1.4 times after
administration of ATP and ATP-H, respectively.
Analysis of the experimental data on changes
in some parameters of plasma and platelet hemo-
TABLE 1. Platelet Aggregation and Anticoagulant Activity of Blood Plasma from Healthy Rats and Animals with Hypofunction
of ACS after Intranasal Administration of ATP and ATP-H
1, 44.5 months
A: administration of NaCl 21.6±2.5 100.0 27.1±1.23 100.0
B: administration of ATP 18.8±3.4 87.1 29.1±1.18 110.0
C: administration of ATPH 11.3±3.1 52.2 30.6±1.35 114.0
2, 88.5 months
A: administration of NaCl 21.0±2.2 100.0 24.8±0.08 100.0
B: administration of ATP 10.9±5.0** 51.2 28.8±0.61* 116.1
C: administration of ATPH 21.3±6.7 100.0 28.8±1.0* 116.0
3; 10 months
A: administration of NaCl 23.9±3.5 100.0 27.1±0.8 100.0
B: administration of ATP 15.5±3.5* 64.8 35.8±0.6* 132.5
C: administration of ATPH 22.4±6.1 97.1 38.4±1.41* 141.0
Note. *
p
<0.05 and **
p
<0.01 compared to the control (subgroup A).
Group, age
%sec%
APTT
ISL
Platelet aggregation
Bulletin of Experimental Biology and Medicine, Vol. 146, No. 3, 2008 PHYSIOLOGY
275
stasis in rats after intranasal administration of ATP
revealed a significant decrease in platelet aggrega-
tion in the blood of rats with normal and, parti-
cularly, with suppressed function of ACS. These
changes became more significant in elder animals.
Administration of ATP-H significantly decreased
platelet aggregation only in healthy animals. In rats
with suppressed function of ACS (8-8.5- and 10
month-old animals), ATP-H did not decrease platelet
aggregation. Administration of ATP-H exhibiting
high anticoagulant activity in vitro [5] is followed by
its release into the blood, which contributes to a signi-
ficant increase in anticoagulant activity. Our results
indicate that intranasal administration of ATP to
animals with suppressed function of ACS and pre-
thrombotic state decreases the degree of prethrom-
bosis by reducing the level of platelet aggregation,
one of the major factors for thrombosis. Intranasal
administration of ATP-H increases anticoagulant
potential of plasma hemostasis in prethrombotic
animals, which decreases the risk of thrombosis.
REFERENCES
1. K. D. Beller, Biol. Med., 12, No. 2, 15-18 (2006).
2. V. V. Dolgov, N. A. Avdeeva, and K. A. Shchetnikovich,
Methods for Study of Hemostasis [in Russian], Moscow (1996).
3. B. A. Kudryashov, Biological Problems of Regulation of the
Liquid State of the Blood [in Russian], Moscow (1975).
4. L. A. Lyapina, W. E. Pastorova, and L. S. Nikolaeva, Izv. Ros.
Akad. Nauk. Ser. Biol., No. 2, 221-225 (2005).
5. L. S. Nikolaeva, V. V. Chirkov, N. A. Dobrynina, et al., Khim.-
Farm. Zh., 39, No. 2, 3-9 (2005).
6. T. Y. Obergan, Byull. Eksp. Biol. Med., 143, No. 3, 267-269 (2007).
7. W. E. Pastorova, L. A. Lyapina, and A. M. Ul’yanov, Izv. Ros.
Akad. Nauk. Ser. Biol., No. 5, 1-4 (2006).
8. S. F. Okado, R. A. Nicholas, S. M. Kreda, et al., J. Biol. Chem.,
281, No. 32, 22,992-23,002 (2006).
9. S. Ricard-Dlum, M. Beraud, N. Raynal, et al., Ibid., 281, No.
35, 25,195-25,204 (2006).
W. E. Pastorova, L. A. Lyapina, et al.
276
0007-4888/08/14630276 © 2008 Springer Science+Business Media, Inc.
Effect of Muramyl Dipeptides on Postsynaptic
GABA, NMDA, and AMPA Receptors and
Presynaptic NMDA Receptors in Rat Brain
V. V. Grigoriev, L. N. Petrova, A. V. Gabreliyan,
and T. A. Ivanova
Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 146, No. 9, pp. 247-249, September, 2008
Original article submitted November 19, 2007
We studied the effect of muramyl dipeptides on postsynaptic GABA, NMDA, and
AMPA receptors and presynaptic NMDA receptors. L,D-Dipeptide more potently than
L,L-dipeptide inhibited postsynaptic NMDA receptors, potentiated GABA and AMPA
receptors, and inhibited 45Ca2+ uptake in synaptosomes from of rat brain cortex. Our
results indicate that muramyl dipeptides modulate function of glutamate and GABA
receptors.
Key Words: muramyl dipeptides; receptors; synaptosomes; calcium
Laboratory for Neurochemistry of Physiologically Active Substances,
Institute of Physiologically Active Substances, Russian Academy
Sciences, Chernogolovka, Moscow region, Russia.
Address for corre
spondence:
grigor@ipac.ac.ru. V. V. Grigoriev
Muramyl dipeptides (MDP), products of degrada-
tion of bacterial peptidoglycan, trigger the multi-
component system of the protective immune re-
sponse in mammals and activation of proinflamma-
tory cytokines [11,13]. They have a toxic effect on
epithelial cells, cause dysfunction of the kidneys
and liver [6], and induce fever and loss of appetite
[5]. Previous studies were performed to evaluate the
direct effect of MDP on some neuroreceptors, in-
cluding serotonin receptors [9]. MDP stimulate the
production of interleukin-1β [7], which has various
effects on the central nervous system (CNS) and
modulates activity of presynaptic NMDA receptors
in rat brain cortex [2]. MDP stereoisomers have
various toxic effects on liver cells. L,D-MDP is
100-fold more toxic than L,L-MDP and D,D-MDP
[8,14].
Here we studied the effect of MDP on postsyn-
aptic GABA, NMDA, and AMPA receptors and
presynaptic NMDA receptors.
MATERIALS AND METHODS
Electrophysiological studies were performed on
freshly isolated cerebellar Purkinje cells and cortical
neurons from rats aging 12-17 and 7-9 days, re-
spectively. These neurons were obtained by the
enzymatic-and-mechanical method. Transmembra-
ne currents in response to application of GABA,
NMDA, or kainic acid (KA) were recorded electro-
physiologically by the whole-cell patch-clamp tech-
nique on an EPC-9 device (HEKA). The substances
were applied by the method of rapid superfusion.
Since AMPA causes strong and rapid desensitiza-
tion of AMPA receptors, activation of these recep-
tors was induced by KK [4].
Cerebral cortical synaptosomes (CCS) were iso-
lated from newborn Wistar rats (9-10 days age) by
the standard method. The brain was homogenized
(glass-Teflon) in a 10-fold volume of cold sucrose
(0.32 M, 900 rpm). The homogenate was centri-
fuged at 1500g for 10 min. The supernatant was
centrifuged at 10,000g for 20 min. For accumu-
lation of the radioactive label, the P2 fraction of
CCS was suspended in incubation buffer A contai-
ning 132 mM NaCl, 5 mM KCl, and 5 mM HEPES
Bulletin of Experimental Biology and Medicine, Vol. 146, No. 3, 2008 PHYSIOLOGY
277
(pH 7.4, protein concentration 1.5-2.0 mg/ml). Cal-
cium concentration in the final volume was 1.25
mM (1.4 µCi/ml). NMDA (200 mM NMDA and 5
µM glycine) was used to stimulate 45Ca2+ uptake
into CCS. After 3-min incubation with NMDA at
37oC, 45Ca2+ uptake was stopped by filtration through
GF/B fiberglass filters (Whatman) with 3-fold wa-
shing with cold buffer B containing 145 mM HEPES,
10 mM Tris, and 54 mM Trilon B (pH 7.4). All
measurements were performed in 4 parallel samples
(4-5 independent experiments). Radioactivity was
measured on a liquid scintillation β-counter. 45Ca2+
uptake into CCS was evaluated as the difference
between the amounts of the label upon stimulation
with NMDA and in the absence of stimulation. This
parameter was expressed as percent of the control
(100%). Specific uptake of 45Ca2+ was calculated as
follows:
K(43/21)=[(Ca4-Ca3)/(Ca2-Ca1)]×100%,
where Ca1 is 45Ca2+ uptake in the control (without
NMDA and MDP); Ca2 is 45Ca2+ uptake in the pre-
sence of NMDA (without MDP); Ca3 is 45Ca2+ up-
take in the presence of MDP (without NMDA); and
Ca4 is 45Ca2+ uptake in the presence of NMDA and
MDP.
The results were analyzed by Student’s t test.
RESULTS
L,D-MDP in various concentrations increased the
amplitude of GABA-induced currents in cerebellar
Purkinje cells. GABA-induced currents increased
most significantly under the influence of L,D-MDP
in concentrations of 10—10-10—8 M (p<0.05). In-
creasing the concentration of L,D-MDP was accom-
panied by an increase in the potentiating effect on
GABA-induced currents. However, these changes
were not monotonous (Fig. 1). L,D-MDP in con-
centrations of 10—13-10—8 M decreased the ampli-
tude of NMDA-induced currents in cortical neu-
rons. Under the influence of L,D-MDP in concentra-
tions of 10—11-10—10 M, the response was 60-70% of
the control (p<0.05, Fig. 2).
MDP stereoisomers inhibited the glutamate-
induced uptake of 45Ca2+ into rat CCS. L,D-MDP
(10—10 M) and L,L-MDP (10—6 M) inhibited 45Ca2+
uptake into CCS by 20%. Hence, MDP have a mo-
dulatory effect on presynaptic glutamate receptors.
Then, we identified the type of receptors affected
by MDP.
45Ca2+ uptake into CCS upon stimulation with
NMDA is related to activation of glutamate NMDA
receptors. NMDA-induced 45Ca2+ uptake into CCS
decreased after treatment with the following anta-
gonists of NMDA receptors: MK-801 (IC50 ~1 µM),
CPP (IC50 ~100 µM), memantine (IC50 ~0.4 µM),
and Mg2+ (IC50 ~100 µM). Our results are consistent
with published data that NMDA activates inotropic
NMDA receptors in the P2 fraction of rat CCS [2].
During stimulation of 45Ca2+ uptake by 200 µM
NMDA (+5 µM glycine), L,D-MDP in concentra-
tions of 10—12-10—8 M had an inhibitory effect on
NMDA-induced 45Ca2+ uptake into CCS. The inhi-
bition of 45Ca2+ uptake into CCS was most signi-
ficant (100%) under the influence of L,D-MDP in
a concentration of 10—10 M (IC50=5×10—12 M). The
dependence of NMDA-induced 45Ca2+ uptake into
CCS on the concentration of L,D-MDP was de-
scribed by a bell-shaped curve (Fig. 3). L,L-MDP
Fig. 1. Effect of L,D-MDP on GABA-induced currents in rat cere-
bellar Purkinje cells.
Fig. 2. Effect of L,D-MDP on NMDA-induced (
1
) and KA-induced
currents in rat cerebral cortical neurons (
2
).
V. V. Grigoriev, L. N. Petrova, et al.
278
in concentrations of 10—11-10—6 M had an inhi-
bitory effect on 45Ca2+ uptake into CCS during sti-
mulation with NMDA. The inhibition of 45Ca2+ up-
take into CCS depended on the concentration of
L,L-MDP. L,L-MDP in concentrations of 10—7-10—6
M was most potent in inhibiting 45Ca2+ uptake into
CCS (IC50=6×10—9 M). Therefore, MDP affect pre-
synaptic NMDA receptors.
L,D-MDP in a concentration of 10—14 M most
significantly inhibited KA-induced currents in hippo-
campal neurons. However, administration of L,D-
MDP in concentrations of 10—13-10—9 M was follo-
wed by an increase in KA-induced currents. L,D-
MDP in a concentration of 10—8 M had an inhibitory
effect on KA-induced currents (Fig. 2).
Our results indicate that L,D-MDP has a mo-
dulatory effect on postsynaptic GABA, NMDA,
and AMPA receptors and presynaptic NMDA re-
ceptors. L,D-MDP potentiates GABA receptors, which
probably contributes to the activating effect on
slow-wave sleep [3]. MDP inhibit NMDA recep-
tors, but potentiate AMPA receptors. These recep-
tors are involved not only in signal transduction in
CNS, but also in the mechanisms of memory and
realization of cognitive functions in the brain [10,
12]. It should be emphasized that L,D-MDP has a
similar effect on presynaptic and postsynaptic NMDA
receptors. Stereoisomers differ in the ability to
modulate presynaptic NMDA receptors. L,D-MDP
was 800-fold more potent than L,L-MDP in inhi-
biting the uptake of 45Ca2+ in CCS.
This work was supported by the International
Science and Technology Center (project No. 2704).
REFERENCES
1. L. N. Petrova and S. O. Bachurin, Byull. Eksp. Biol. Med., 142,
No. 7, 51-54 (2006).
2. L. N. Petrova, V. V. Grigoriev, and S. O. Bachurin, Ibid., 140,
No. 12, 645-646 (2005).
3. J. Andren, K. Andrews, L. Brown, et al., Behav. Brain Res.,
69, Nos. 1-2, 85-90 (1995).
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Fig. 3. Effects of L,D-MDP (
1
) and L,L-MDP (
2
) on 45Ca2+ uptake
into rat CCS upon stimulation with NMDA.
Bulletin of Experimental Biology and Medicine, Vol. 146, No. 3, 2008 PHYSIOLOGY
279
0007-4888/08/14630279 © 2008 Springer Science+Business Media, Inc.
GENERAL PATHOLOGY AND PATHOPHYSIOLOGY
Cytomorphological Study of the Development
of Fibrotic Complications in Chronic SiO2
Granulomatosis in the Liver during Radon
Treatment
M. S. Novikova, O. V. Potapova, and V. A. Shkurupy
Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 146, No. 9, pp. 250-253, September, 2008
Original article submitted May 20, 2008
Granulomatosis was induced in male Wistar rats by intravenous injection of SiO2. The
course of SiO2 granulomatosis was cyclic in animals receiving radon baths: phase 1 was
characterized by an increase in the number and size of granulomas, while during phase
2 the intensity of the inflammatory process decreased and fibrosing of granulomas
progressed. No trends to alleviation of the inflammatory process were noted in rats with
SiO2 granulocytosis receiving tap water baths.
Key Words: silicon dioxide; radon; granulomatous inflammation; fibrosis; liver
Research Center of Clinical and Experimental Medicine, Siberian
Division of Russian Academy of Medical Sciences, Novosibirsk,
Russia.
Address for correspondence:
marnovcom@yandex.ru. M. S.
Novikova
The nature of positive effects of radon exposure
[2,7] is still disputed; radon procedures are mainly
used in patients with destructive processes in the
locomotor system and chronic inflammatory pro-
cesses, one of the complications of which is ex-
cessive formation of fibrous tissue. The model of
chronic granulomatous inflammation induced by
injection of SiO2 can be used for studies of fibrosis
and interrelationships between cells largely determi-
ning this process in any organ: lymphocytes, ma-
crophages, and fibroblasts [9]. High clearing activity
of the liver towards microcorpuscular objects due to
high concentration of resident macrophages (Kupffer
cells) in it [9] makes this organ convenient for studies
of fibrotic processes induced by injection of a lyso-
somotropic factor, for example, SiO2 [9].
We studied the effect of radon on the develop-
ment of chronic SiO2 granulomatosis and its com-
plication (fibrosis).
MATERIALS AND METHODS
The study was carried out on 36 male Wistar rats
(300-350 g) in which granulomatosis was induced
by a single injection (into the caudal vein) of 0.8
ml of microcrystal SiO2 suspension (1-5-µ particles)
in 0.85% NaCl (SiO2 dose 35 mg/kg).
The animals were divided into 3 groups (12 per
group): group 1 consisted of untreated rats; groups
2 and 3 rats received tap water baths and baths of
silicon nitrate radon-containing water from Belo-
kurikha Health Resort, respectively (3, 10, or 20 daily
baths). The animals were plunged completely, ex-
cept the heads, in water (22-24oC) for 30 min;
radon concentration was 8 nCi/liter.
Material for analysis was collected on days 3,
10, and 20 after the start of balneotherapy, which
Bulletin of Experimental Biology and Medicine, Vol. 146, No. 3, September, 2008
280
corresponded to days 13, 20, and 30 after SiO2
injection. The animals were sacrificed by deca-
pitation under ether narcosis.
Liver specimens for light microscopy were stai-
ned with hematoxylin and eosin and by van Gieson
method. The following parameters were studied:
density (Nai) and diameters (in µ) of SiO2 granulo-
mas in the liver, cell composition of granulomas
(percent from the total number of cells in the granu-
loma), volume density (Vv) of fibrous connective
tissue in granulomas, fibroplastic “activity” of fib-
roblasts, which was expressed through Vv of colla-
gen fibers per fibroblast in the granuloma, fibrous
tissue concentration in the organ (product of fib-
rous tissue Vv in granulomas and Nai of granulomas
in the organ).
The significance of differences between the
means was evaluated using Student’s t test, the dif-
ferences were considered significant at p<0.05 [1,11].
RESULTS
Granulomas formed in the livers of animals of all
groups on day 13 after SiO2 injection (day 3 after
the start of balneotherapy). The Nai was maximum
in the livers of groups 1 and 2 rats (Table 1); the
TABLE 1. Morphometric Parameters of Structural Changes in the Liver of Animals with Chronic SiO2 Granulomatosis after
Balneotherapy (
M
±
m
)
Nai 3 0.520±0.073 0.450±0.015 0.300±0.025+о
10 0.65±0.04 0.500±0.017+0.800±0.038*+
20 0.720±0.061 0.630±0.056* 0.710±0.049
Diameter of granulomas, μ
3 29.53±0.74 30.52±1.07 25.08±1.15+о
10 35.58±1.05* 34.27±1.26* 50.28±0.87*+
20 36.95±1.05 37.18±0.91* 35.40±1.56
Cell composition of granulomas, %
macrophages 3 87.40±1.90 85.40±0.77 71.78±1.29+о
10 87.90±0.89 84.12±1.54+70.15±2.04о
20 84.30±1.19 84.31±1.33 75.17±1.62*+о
lymphocytes 3 5.25±0.025 6.14±0.043 13.43±0.63+
10 4.100±0.029 6.250±0.052 10.21±0.15+
20 4.760±0.037* 5.450±0.063 3.500±0.038+о
neutrophils 3 2.180±0.018 1.61±0.03+7.430±0.071+
10 2.280±0.016* 1.980±0.017* 6.400±0.025+
20 1.96±0.03 1.79±0.08 1.19±0.07+о
fibroblasts 3 5.170±0.074 6.850±0.036 7.360±0.067+
10 5.720±0.062* 7.650±0.075* 13.24±0.96*+
20 8.980±0.053* 8.450±0.048*+20.14±1.32*+
Fibrous tissue Vv in granuloma, % 3 5.57±0.45 7.12±0.61 8.25±0.42+
10 4.95±0.27 7.75±0.45 16.75±0.85*+
20 8.02±0.31* 8.20±0.58 22.84±1.26*+
Fibroplastic “activity” of fibroblasts 3 1.07±0.10 1.03±0.04 1.12±0.08
10 0.90±0.07 1.010±0.015 1.17±0.10
20 0.89±0.05 0.970±0.045 1.130±0.075
Fibrous tissue concentration
coefficient in organ 3 2.90±0.09 3.20±0.10 2.46±0.07+о
10 3.22±0.17 3.86±0.22* 13.40±0.34*+
20 5.77±0.36* 5.17±0.43* 16.18±0.59*+
Note.
p
<0.05 compared to: *previous term, +group 1, ogroup 2.
Parameter Group 3
Balneotherapy
duration, days Group 1 Group 2
Bulletin of Experimental Biology and Medicine, Vol. 146, No. 3, 2008 GENERAL PATHOLOGY AND PATHOPHYSIOLOGY
281
granulomas consisted mainly of macrophages, few
lymphocytes and fibroblasts (Table 1). After 10
days of balneotherapy (day 23 after SiO2 injection),
the granuloma Nai in the liver increased in animals
of all groups (Table 1). In group 3, this value was
by 23% higher than in group 1 and 60% higher
than in group 2. After 20 days of balneotherapy
(day 33 after SiO2 injection), Nai decreased by 10%
only in group 3 rats and virtually did not differ from
the value in animals of other groups (Table 1).
Granulomas increased in size on day 10 of
balneotherapy in animals of all groups. In com-
parison with the previous term (day 3), enlargement
of granulomas in groups 1 and 2 was negligible,
while in group 3 this parameter increased 2-fold
and was 1.4 times higher than in other groups (Ta-
ble 1). On day 20, granulomas in the livers of group
3 rats decreased by 30% in comparison with the
previous term and their size virtually did not differ
from that in rats of other groups (Table 1).
On day 10 of balneotherapy, the number and
size of granulomas in the livers of group 3 rats
were significantly higher than in other groups, pre-
sumably due to the prooxidant effect of radon [3,
10] and activation of free radical oxidative pro-
cesses. It is known that less than 1% radon present
in radon water penetrates into the body after radon
baths, while the greater part (60%) of radon dif-
fuses into the derma [7]. Radon present in thermal
waters releases α-particles, which stimulate the for-
mation of superoxide radicals and peroxide com-
pounds in the derma [7]; in addition, radiogenic
radicals accumulate with subsequent oxidation of
a variety of biosubstrates at the molecular level
[10]. This leads to modification of the serum pro-
and antioxidant activities [6,8,10], which serves as
an indicator of the modulatory effect of radon, pri-
marily on macrophages. Serum prooxidant activity
increases during the early period of balneotherapy
(day 3) [6,10], which under conditions of a gra-
nulomatous inflammation can be caused by macro-
phage activation and enhanced production of acti-
vated oxygen metabolites, proinflammatory cyto-
kines chemoattractants [4,6,10]. This leads to intense
migration of mononuclears and other immunocom-
petent cells (including lymphocytes) into the focus
of inflammation, which presumably explains en-
largement of granulomas on day 10. On day 20 of
balneotherapy (group 3), the increase in serum
antioxidant activity [6,10] was paralleled by macro-
phage activation with increased secretion of anti-
inflammatory cytokines (IL-1, IL-10) [8] and atte-
nuation of the inflammatory process [6,10].
On day 20 of balneotherapy, the specific cel-
lular reaction under the effect of radon degradation
products in liver granulomas in group 3 rats mani-
fested by a 3-fold reduction in lymphocyte count
in comparison with the previous term (day 10),
which was paralleled by a 50% increase in fibro-
blast number (Table 1) and formation of fibrosing
granulomas. Fibrous tissue Vv in liver granulomas
of groups 1 and 2 rats was 2.35 times lower than
in group 3, while fibroplastic “activity” of fibro-
blasts was virtually the same in all groups during
all periods of observation (Table 1). The coefficient
of fibrous tissue concentration in the organ in the
radon therapy group was significantly higher on
day 10 (4-fold higher than in groups 1 and 2) and
on day 20 (3-fold higher than in groups 1 and 2).
According to the results of an analogous experi-
ment, radon baths modify connective tissue meta-
bolism, which can be seen from a reduction of
fibronectin level and increased concentration of
glycosaminoglycans in the sera of animals with
SiO2 granulomatosis [5]. Interleukin-4 and IL-13
stimulate macrophage secretion of arginase 1, re-
sponsible for the synthesis of polyamines and prol-
ine, essential for fibroblast proliferation and colla-
gen production [4]. The results indicate that bal-
neotherapy therapy with radon-containing water
reduces the severity of acute inflammation pre-
sumably due to improvement of the antioxidant
potential at the expense of a sort of a “training”
effect of antioxidant systems [10], and stimulate the
processes of reparative regeneration of the connec-
tive tissue, manifesting in increased proliferative,
but not synthetic activity of fibroblasts. This can be
useful in such diseases as osteochondrosis, verte-
bral instability in distortions of different genesis.
These data necessitate more strict definition of
indications for radon therapy in other diseases, as
its negative effect in chronic inflammatory proces-
ses, for example, in the lungs cannot be ruled out.
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M. S. Novikova, O. V. Potapova, and V. A. Shkurupy
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thods in Medical Research, Oxford (2002).
Bulletin of Experimental Biology and Medicine, Vol. 146, No. 3, 2008 GENERAL PATHOLOGY AND PATHOPHYSIOLOGY
283
0007-4888/08/14630283 © 2008 Springer Science+Business Media, Inc.
Free Radical Lipid Peroxidation during Amiodarone
Therapy for Postinfarction Cardiosclerosis
T. Yu. Rebrova and S. A. Afanasyev
Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 146, No. 9, pp. 254-256, September, 2008
Original article submitted February 26, 2008
Postinfarction remodeling of the heart in animals with myocardial infarction was accom-
panied by activation of lipid peroxidation in myocardial tissue and blood serum and
decrease in antioxidant enzyme activity. In animals with postinfarction cardiosclerosis
treated with amiodarone, we observed decreased accumulation of lipid peroxidation
products and normalization of superoxide dismutase activity.
Key Words: lipid peroxidation; amiodarone; postinfarction cardiosclerosis
Institute of Cardiology, Tomsk Research Center, Siberian Division of
the Russian Academy of Medical Sciences, Tomsk, Russia.
Address
for correspondence:
rebrova@cardio.tsu.ru. T. Yu. Rebrova.
The development of postinfarction cardiosclerosis
(PICS) is accompanied by structural and electro-
physiological remodeling of the myocardium. Elec-
trophysiological remodeling of the myocardium is
a complex of molecular, metabolic, and ultrastruc-
tural changes in cardiomyocytes, which determines
electrophysiological dysfunction of the heart and
development of arrhythmias [5]. These features con-
tribute to high risk of paroxysmal arrhythmias and
sudden cardiac death in the postinfarction period
[4]. Myocardial infarction and PICS are followed
by activation of free radical processes in the myo-
cardium [8,11], which impairs the integrity of car-
diomyocyte membranes and causes dysfunction of
Ca2+-ATPase. These processes result in contractile
dysfunction and change in energy supply to the
myocardium [9,2]. Variations in intracellular ener-
gy metabolism are accompanied by changes in Na+,
K+, and Ca2+ currents and, therefore, disturbances
in automatism and conduction in cardiac cells [2].
Class III antiarrhythmic drugs are widely used
for the correction of cardiac arrhythmias. One of
these drugs is amiodarone (Cordarone). Amioda-
rone is used in clinical practice for the treatment of
severe arrhythmias [10]. Multicenter, randomized,
placebo-controlled trials (CAMIAT and EMIAT)
showed that amiodarone therapy significantly de-
creased the risk of arrhythmic and cardiac death in
postinfarction patients with ventricular extrasystoles
[11,13]. Much recent attention is paid to the me-
chanisms for action of amiodarone on ischemic myo-
cardium during PICS.
Here we studied the intensity of lipid peroxi-
dation (LPO) in myocardial tissue and blood serum
from rats with PICS, which received the course of
amiodarone therapy.
MATERIALS AND METHODS
Experiments were performed on 30 Wistar rats weig-
hing 180-200 g. Groups 1 (control 1) and 2 (con-
trol 2) consisted of sham-operated animals and rats
with PICS, respectively. Group 3 animals with PICS
receiving amiodarone.
The development of cardiosclerosis in group 2
and 3 rats was triggered by myocardial infarction
caused by coronary occlusion [12]. Thoracotomy
was performed in ether-anesthetized animals under
sterile conditions. A ligature was applied to the
upper third of the left descending coronary artery.
The surgical wound was sutured. The animals
were maintained in a vivarium for 45 days under
standard conditions. PICS was observed by the end
of this period.
Bulletin of Experimental Biology and Medicine, Vol. 146, No. 3, 2008 GENERAL PATHOLOGY AND PATHOPHYSIOLOGY
284
Distilled water was administered through a gas-
tric tube to group 1 and 2 rats (twice a day, 14
days) starting from the 30th day after surgery. Group
3 animals intragastrically received amiodarone in a
dose of 10 mg/kg (daily dose 20 mg/kg).
Control and treated animals were examined af-
ter 45 days. Blood samples were centrifuged at
3000 rpm for 5 min. Serum aliquots were stored in
liquid nitrogen. The samples of myocardial tissue
were taken.
The intensity of LPO in blood serum and myo-
cardial tissue was estimated from the contents of
conjugated dienes (CD) [3] and malonic dialdehyde
[6] and activities of antioxidant enzymes super-
oxide dismutase [1] and catalase [7].
The results were analyzed by paired Student’s
t test. The differences were significant at p<0.05.
RESULTS
Postinfarction remodeling of the myocardium in
group 1 animals was accompanied by pronounced
activation of free radical processes in the myo-
cardial tissue. The contents of CD and MDA in
myocardial homogenates from PICS rats were hig-
her than in sham-operated animals (by 1.3 and 1.8
times, respectively; Table 1).
The development of PICS in rats was accom-
panied by an increase in the contents of MDA and
CD in blood serum (by 22 and 90%, respectively,
compared to sham-operated animals; Table 2). These
changes reflect activation of LPO during postin-
farction remodeling of the myocardium.
The course of intragastric administration of amio-
darone for 2 weeks was followed by a decrease in
CD content in the myocardium of group 3 rats
compared to group 2 animals. MDA content in group
3 rats was lower than in group 2 animals. However,
MDA content in group 3 rats remained higher than
in sham-operated animals.
Similar inhibition of LPO was also observed in
blood serum of group 3 rats. Amiodarone decrea-
sed the contents of MDA and CD in group 3 ani-
mals by 37 and 59%, respectively, compared to
group 2 rats (Table 2).
These data show that the course of intragastric
administration of amiodarone decreased accumula-
tion of LPO products in PICS animals. These chan-
ges probably contribute to the antiarrhythmic effect
during postinfarction remodeling of the myocar-
dium.
Components of the endogenous antioxidant
system are the major factors for cell protection
from reactive oxygen species and lipid hydroper-
oxides. Therefore, the effectiveness of myocardial
recovery after coronary disease can be estimated
from antioxidant enzyme activity in the myocar-
dium and blood serum of animals with postinfarc-
tion remodeling of the myocardium. The effect of
amiodarone on the antioxidant system under these
conditions is also of considerable diagnostic signi-
ficance.
Measuring of antioxidant enzyme activity in
myocardial samples and blood serum from animals
with PICS showed that the development of cardio-
sclerosis was accompanied by a significant decrease
TABLE 2. Effect of the Course of Amiodarone Administration on LPO and Antioxidant Enzyme Activity in Blood Serum from
Rats with PICS (
M
±
m
)
1 1.03±0.08 20.99±2.03 20.52±1.63 0.86±0.05
2 1.96±0.09* 25.58±2.26** 15.92±0.95** 0.09±0.01***
3 1.17±0.12+19.22±1.31++ 11.48±0.64***++ 0.24±0.02***++
Group SOD, mmol/mg
protein/min
Catalase, mmol/g
protein/min
CD,
ΔE232/g tissue
MDA,
nmol/g tissue
TABLE 1. Effect of the Course of Amiodarone Administration on the Content of LPO Products and Antioxidant Enzyme
Activity in the Myocardium of Rats with PICS (
M
±
m
)
1 0.32±0.03 9.48±0.85 30.23±2.64 0.87±0.04
2 0.43±0.04* 16.63±0.97** 18.10±1.07** 0.09±0.01***
3 0.34±0.03+11.23±0.89*++ 21.31±1.06**+0.19±0.01**++
Note. Here and in Table 2: *
p
<0.05, **
p
<0.01, and ***
p
<0.001 compared to group 1; +
p
<0.05 and ++
p
<0.01 compared to group 2.
Group SOD, mmol/mg
protein/min
Catalase, mmol/g
protein/min
CD,
ΔE232/g tissue
MDA,
nmol/g tissue
Bulletin of Experimental Biology and Medicine, Vol. 146, No. 3, 2008 GENERAL PATHOLOGY AND PATHOPHYSIOLOGY
285
in catalase and SOD activities (compared to sham-
operated animals; Tables 1 and 2).
The decrease in SOD and catalase activities du-
ring experimental cardiosclerosis is probably rela-
ted to inhibition of their active centers by fatty acid
peroxides. The formation of these compounds in-
creases in myocardial ischemia [9]. On the other
hand, the decrease in antioxidant enzyme activity
induced by various factors contributes to the in-
creased formation of LPO products, which is me-
diated by the feedback mechanism.
Activities of catalase and SOD in the myocar-
dium of group 3 animals increased compared to
those in group 2 rats (Table 1). However, antioxi-
dant enzyme activity in these animals remained
lower than in sham-operated rats.
Serum activities of catalase and SOD in animals
with experimental cardiosclerosis were also lower
than in sham-operated rats.
SOD activity in group 3 rats was higher than
in group 2 animals (Table 2). At the same time,
catalase activity decreased in group 2 rats. The
mechanism underlying the decrease in serum cata-
lase activity remains unclear and requires further
investigation.
Our results indicate that experimental postin-
farction remodeling of the heart is accompanied by
activation of LPO and decrease in antioxidant en-
zyme activity in myocardial tissue and blood se-
rum. The course of amiodarone administration in a
daily dose of 20 mg/kg was followed by a decrease
in the contents of MDA and CD in myocardial tis-
sue and blood serum and contributed to the increase
in SOD and catalase activities. Amiodarone-indu-
ced decrease in LPO during postinfarction cardiac
remodeling is associated with the effect of the test
drug on ion transport in myocardial cells. These
changes promote normalization of the heart rate
and decrease in oxygen demands of the myocar-
dium. Published data show that administration of
amiodarone to animals with dilated cardiomyopathy
has a cardioprotective effect. This drug prevents
irreversible degeneration of cardiomyocytes, im-
proves the dynamics of oxidation-reduction enzyme
activities, and normalizes bioelectric activity of the
heart and total content of adenyl nucleotides [12].
Amiodarone probably has a similar effect under con-
ditions of PICS and contributes to inhibition of LPO.
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T. Yu. Rebrova and S. A. Afanasyev
286
0007-4888/08/14630286 © 2008 Springer Science+Business Media, Inc.
Blood Lymphocyte Proliferation Reaction
in Autoimmune Thyropathies
O. I. Urazova, E. B. Kravets, Yu. V. Nedosekova,
A. V. Rogaleva, V. N. Kuznecova, and O. A. Rumpel
Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 146, No. 9, pp. 257-260, September, 2008
Original article submitted December 5, 2007
Spontaneous and stimulated reaction of lymphocyte proliferation and blood levels of
CD25- and CD120-expressing lymphocytes were studied in patients with autoimmune
thyroiditis and diffuse toxic goiter. Activation of CD25- and CD120-presenting function
of blood lymphocytes was revealed. The findings indicate that the course of autoimmune
thyroiditis and diffuse toxic goiter is associated with increased levels of basal, mitogen-
and cytokine-stimulated proliferation of blood lymphocytes, particularly pronounced in
thyroiditis in the euthyrosis stage.
Key Words: lymphocyte proliferation reaction; autoimmune thyroiditis; diffuse toxic
goiter
Siberian State Medical University, Russian Ministry of Health, Tomsk,
Russia.
Address for correspondence:
nedosekova@yandex.ru. Yu.
V. Nedosekova
Autoimmune thyropathies (AITP) autoimmune thy-
roiditis (AIT) and diffuse toxic goiter (DTG)) are
among the most incident diseases in the structure
of thyroid abnormalities. Despite numerous studies
of this problem, the mechanism of pathogenetic
reactions in AITP remains unknown [3,7].
Autoimmune thyropathies belong to a group of
complex polygenic diseases, their development is
determined by numerous genetic, endo- and exo-
genous factors provoking the antithyroid immune
reaction manifesting in high production of autoanti-
bodies to components of thyrocyte membrane and
thyroid hormones (thyroglobulin, thyroperoxidase,
thyrotropin receptor, etc.) [1,7,9]. In addition, imba-
lance between the pro- and antiinflammatory cyto-
kines leads to changes in the immune homeostasis,
primarily to changes in proliferative activity of im-
munocompetent cells [2,3]. Impairment of B and T
lymphocyte proliferation processes is an important
component in the progress of destructive and in-
flammatory processes in the thyroid tissue.
We evaluated proliferative activity of periphe-
ral blood lymphocytes in AITP.
MATERIALS AND METHODS
Sixty patients with AITP aged 19-55 years (mean
age 42.1±3.9 years) were examined. The diagnosis
was in each case verified with consideration for
clinical picture and paraclinical parameters in ac-
cordance with routine criteria [1]. The patients were
divided into 3 groups in accordance with the noso-
logical entities and functional activity of the thy-
roid. Group 1 consisted of 20 patients with AIT in
a state of euthyrosis (hormonal profiles within the
norm); group 2 consisted of 16 patients with AIT
with thyroid hypofunction (mean level of thyro-
tropin 10.59±3.25 mMI/liter), the concentration o