P.1.g.056
Effects of antipsychotics on the respiratory chain and NADPH oxidase in
the brain of phencyclidine perinatally treated rats
Nataša Petronijević, Gordana Jevtić, Milica Živković, Milica Velimirović, Tatjana Nikolić, Tihomir Stojković,
Mikan Lazović
Institute of Medical and Clinical Biochemistry, School of medicine, University of Belgrade, Serbia
INTRODUCTION
Perinatal phencyclidine (PCP) administration to rodents represents an actual animal model of schizophrenia (SCH). Oxidative stress has been documented in
SCH patients and recent studies suggest that this could be a consequence of antipsychotic medication. Mitochondria have been recognized as the most
important source of free radicals but lately the role of NADPH oxidase (NOX2) enzymes in redox disregulation has been suggested. n
The aim of this study was to elucidate the
effects of haloperidol (typical antipsychotic)
and clozapine (atypical antipsychotic) on the
activity of respiratory chain enzymes
(complex I and IV), and protein expression
of gp91phox, p22phox, p67phox and p47phox
NOX2 subunits in the cortex and
hippocampus of PCP perinatally treated rats.
NOX2, a superoxide-producing enzyme, is
composed of membrane (gp91phox and
p22phox) and cytosolic regulatory (p47phox,
p67phox and p40phox) subunits. NOX2
activation requires translocation of cytosolic
subunits and activated Rac1/2 (small
GTPase) to the membrane.
Spiers et al. Front Neurosci 2015, 8:456
MATERIAL AND METHODS
Male Wistar rats were divided into six groups and subcutaneously treated on 2nd, 6th, 9th and 12th postnatal
(P) day, either with PCP (10mg/kg) or NaCl. From P35 to P100, one NaCl (NaCI-H) and one PCP (PCP-H)
group have received haloperidol (1mg/kg/day), one NaCl (NaCl-C) and one PCP (PCP-C) groups have
received clozapine (20mg/kg/day) dissolved in drinking water. The remaining NaCl (control) and PCP groups
have received drinking water. Animals were sacrificed on P100. Enzyme activity was determined
spectrofotometrically, while protein expression was determined by Western blot.
RESULTS
**
*
80
60
40
20
0
** **
##
180
Density ratio p67
phox
/Actin (%)
160
*
120
120
#
#
#
60
40
20
Cortex
100
**
##
80
60
40
20
0
Hippocampus
NaCl
PCP
NaCl-H
PCP-H
NaCl-C
PCP-C
Cortex
Hippocampus
NaCl
PCP
NaCl-H
PCP-H
NaCl-C
PCP-C
##
140
*
*
**
#
*#
80
0
Hippocampus
Cortex
120
100
#
#
140
/Actin (%)
*
**
phox
* *
NaCl
PCP
NaCl-H
PCP-H
NaCl-C
PCP-C
140
/Actin (%)
##
120
100
*
phox
##
Density ratio p22
Density ratio gp91
phox
/Actin (%)
140
NaCl
PCP
NaCl-H
PCP-H
NaCl-C
PCP-C
Density ratio p47
**
* *
Activity of complex I and COX in mitochondrial
fraction of NaCl group, PCP group and groups
treated with haloperidol (NaCl-H and PCP-H) and
clozapine (NaCl-C and PCP-C) in the cortex and
hippocampus of adult animals. Results are
presented as the mean±SEM.
100
80
60
NaCl
(n=6)
PCP
(n=6)
Cortex
24.5±4.9
22.3±4.1
Hippocampus
23.5±4.2
22.8±4.0
Complex I
(U/mg protein)
NaCl-H
(n=6)
PCP-H
(n=6)
NaCl-C
(n=6)
PCP-C
(n=6)
26.0±1.8
29.2±2.5
27.8±1.8
23.4±2.2
23.6±3.8
27.3±1.1
24.7±2.3
28.4±5.5
40
20
0
Cortex
NaCl
(n=6)
Hippocampus
*p<0.05 ; **p<0.0
1– compared to NaCl group
#p<0.05 ; ##p<0.01 - compared to PCP group
COX
(mmol/mg protein/60min)
PCP
NaCl-H
(n=6)
(n=6)
PCP-H
(n=6)
NaCl-C
(n=6)
PCP-C
(n=6)
Cortex
13.4±1.9
11.4±1.3
11.8±0.7
9.5±0.8*
9.8±0.89
7.3±1.0*
Hippocampus
11. 2±2.7
8.2±0.8
10.1±2.8
11.0±1.41
7.2±1.4
6.0±1.9*
CONCLUSION
Obtained data suggest that PCP administration and antipsychotic medication induce alteration of different
subunits forming NOX2 complex in both investigated brain structures. Our findings are indicative for the
involvement of NOX2 in free radicals generation in this animal model of SCH. Opposite and even more
pronounced changes noticed in clozapine treated groups, especially in cortical regions, suggest that
clozapine treatment could be more harmful for the brain cells. Further investigations of the role of NOX2 in
pathophysiology of SCH and its potential use as a therapeutic target are needed.
No potential conflict of interest