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Iqbal et al. European Journal

Europeanof Biomedical
Journal
ISSN 2349-8870
of Biomedical and Pharmaceutical Sciences
Volume: 6
AND Pharmaceutical sciences Issue: 8
443-450
http://www.ejbps.com Year: 2019

PREGNENOLONE SULPHATE REVERSED KETAMINE INDUCED BEHAVIORAL

CHANGES IN RATS

Mohd. Rashid Iqbal1*, Sudhir Mishra1 and Yam Nath Paudel2

1
*Department of Pharmacology, IIMT College of Pharmacy, Greater Noida U.P, India.
2
Neuropharmacology Research Laboratory, Jeffrey Cheah School of Medicine and Health Sciences, Monash University
Malaysia, Bandar Sunway, Selangor, Malaysia.

*Corresponding Author: Mohd. Rashid Iqbal

Department of Pharmacology, IIMT College of Pharmacy, Greater Noida U.P, India.

Article Received on 20/06/2019 Article Revised on 10/07/2019 Article Accepted on 30/07/2019

ABSTRACT
Pregnenolone and its derivatives have been implicated in neuroprotection and enhance NMDA receptor
neurotransmission pointing out their therapeutic potential in schizophrenia. This study was aimed to evaluate the
effects of pregnelonone sulphate (Preg S) on behavioural changes and markers of oxidative stress induced by
ketamine (Ket) in rats. All rats received intraperitoneal (i.p) injection daily for 14 days with different drug
treatments viz. Preg S (11.86 mg/kg), risperidone (2 mg/kg/day) alone and in combination with ketamine (30
mg/kg). Different behavioural tests which included elevated plus maze, spontaneous alternation behaviour,
locomotor activity tests were performed after 24 hrs of last dosing followed by the estimation of markers of
oxidative stress and acetylcholine esterase (AchE) activity in rat’s brain. Ket produced significant changes in
behaviour resembling to that of negative symptoms and memory impairment seen in schizophrenic patients. Preg
S treated rats spent more time in open arm in elevated plus maze test, decreased percentage alternation and
improved locomotor activity as compared to Ket treated rats. The levels of antioxidants such as superoxide
dismutase (SOD) and reduced glutathione (GSH) were reduced while thiobarbituric acid reactive substances
(TBARS) were elevated. Preg S administration normalized the antioxidant parameters. Preg S improved learning
and memory in elevated plus maze, spontaneous alternation behaviour, locomotor activity, decreased AChE
activity and reduced oxidative stress. Thus, we may conclude that Preg S offered neuroprotective and antioxidant
effect in Ket induced behavioural symptoms.

KEYWORDS: Pregnenolone sulphate, ketamine, neuroprotective, antioxidant, schizophrenia.

1. INTRODUCTION these neurosteroids are metabolized in the brain from

Schizophrenia (SCZ) is a major mental illness affecting precursor compounds originating from endocrine
approximately 1% population of world, responsible for sources. These neurosteroids are synthesized de novo in
causing the changes in perception, thoughts and the brain from cholesterol.[8,9] Neuroactive steroids are
behaviour.[1] SCZ is characterized by a wide range of essential for the proper development and functioning of
symptoms which includes positive symptoms, negative the adult brain and play a major role in the stress
symptoms, cognitive and neuropsychological response.[10] Based on plasma and cerebral spinal fluid
dysfunction and mood symptoms.[2,3] In addition to the (CSF) level studies in humans and preclinical evaluation
deficits in the cognitive function like attention and of drugs on brain and plasma levels in laboratory
memory, it is the leading cause of suicide along with animals, it was demonstrated that these steroids may
anxiety and depression in about 10% cases.[4] contribute to the pathology and symptoms of some
psychiatric illnesses and their levels may be affected by
Despite the availability of better anti-psychotic drugs, drugs used to treat these disorders.[11-14]
complete cure for SCZ has been elusive, and research is
continuing for novel anti-psychotics possessing unique Neurosteroids, such as dihydroepiandrosterone (DHEA),
pharmacological profile to take care of positive, negative pregnenolone and their derivatives have been implicated
symptoms as well as cognitive deficits.[5-7] in neuroprotection and enhancement of NMDA receptor
neurotransmission suggesting therapeutic potential in
A number of steroid hormones are synthesized in the SCZ, possibly by actions at sigma1 receptors.[15-17]
brain which exists in higher concentrations in the Pregnenolone sulphate (Preg S) is a neurosteroid with
nervous system than in the plasma. It is now known that excitatory effects in the brain, acting as a potent negative

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Iqbal et al. European Journal of Biomedical and Pharmaceutical Sciences

allosteric modulator of the GABAA receptor and a symptoms like locomotor activity changes seen in
weak positive allosteric modulator of the NMDA elevated plus maze.[25]
receptor and agonist of the sigma receptor.[17-19]
2.3. Treatment schedule
With respect to SCZ, Pregnenolone actions on learning Wistar albino rats of either sex were used in the present
and memory in rodent models have been studied as study. The duration of study was 14 days. The Animals
cognitive symptoms were improved and it may be were divided randomly into 6 groups each containing of
hypothesized that their long-term treatment may improve 6 animals. Group-I Normal Control, Group-II Ketamine,
the outcome and quality of life in patients with SCZ.[20,21] Group-III, Preg S per se, Group-IV Risperidone per se,
The neurosteroid pregnenolone may represent a Group-V Preg S + Ketamine, Group-VI Risperidone +
promising and mechanistically novel agent for cognitive Ketamine. In group I, rats were administered normal
and negative symptoms in SCZ. Pregnenolone and its saline (1 ml/kg body weight, i.p). Group II, Ketamine (30
sulphated derivative, Preg S enhance learning and mg/kg/d, i.p), Group III, received Preg S per se (11.86
memory in animal models at concentrations that are mg/kg/d, i.p. dissolved in 0.9% NaCl), Group IV,
physiologically relevant and known to be present in received Risperidone Per se (2 mg/kg/day, i.p), Group V,
human brain.[22-24] received Preg S (11.86 mg/kg, i.p) + Ketamine (30
mg/kg i.p) and Group-VI- Risperidone (2 mg/kg/day, i.p)
In lab animals, the symptoms of SCZ are produced by + Ketamine (30 mg/kg i.p) respectively.
various techniques including use of chemicals etc.
Becker et al reported that sub-chronic treatment of After 24 hrs of last dosing, the behavioural tests were
Ketamine (Ket) (30 mg/kg, i.p) induced changes in rat carried out followed by the estimation of markers of
behaviour resembling to that of the symptoms of SCZ[25] oxidative stress and acetylcholine esterase activity in
and this model was employed in the present research rat’s brain.
study to observe the schizophrenic behavioural
symptoms of animals. Further, the role of oxidative 2.4. Behavioural tests
stress has been implicated in the pathophysiology of 2.4.1. Spontaneous Alternation Behaviour (SAB)
SCZ[26-28] thus, the present study was carried out to This method was used to study the effect of drugs on
determine the effect of Preg S on behavioural changes learning and memory. Spontaneous alternation behaviour
and markers of oxidative stress in Ket induced (SAB) was performed in a plus maze to assess effect of
behavioural changes in rats resembling to schizophrenic drugs on short term memory with respect to spatial
symptoms in humans. orientation and perception as per earlier reported
method.[30] The animals were placed in plus maze. The
2.0. MATERIALS AND METHODS maze (85 cm height) was constructed of wood painted
2.1. Animals grey and contained a central platform (25 cm diameter),
Wistar albino rats, weighing 200-230 gm, were procured from which radiated four symmetrical arms (55cm long×
from the Central Animal House Facility, Hamdard 10 cm wide), with 12 cm wall. After being placed in the
University, New Delhi, India. The animals were kept in central platform, rats were allowed to traverse the maze
polypropylene cages under standard laboratory freely for 12 min. The number and sequence of entries
conditions (12 hours’ light/dark cycles) and had free were recorded. An alternation was defined as entry into
access to a commercial pellet diet and water ad libitum. four different arms on an overlapping quintuple set. Five
The animal house temperature was maintained at 25 ±2 consecutive arm choices within the total set of arms
ºC. This study was approved by the Institutional Animal choices constitute a quintuple set. A quintuple consisting
Ethics Committee (IAEC) [Reg. No1115] Jamia of arm choices A, B, A, C, D was considered as an
Hamdard, New Delhi on dated 25 Feb 2016. alternation, while the set with A, B, A, C, B did not.
Using this procedure, percentage alternation is equal to
2.2. Preparation and drug Administration the ratio of actual alternation to possible alternation ×
All the drugs solutions were administered 100. Possible alternation sequences are equal to the no of
intraperitoneally (i.p) for 14 days to rats. Preg S was arms entries minus 4.
procured from Sigma Aldrich Bangalore and was
administered at a dose of 11.86 mg/kg/day, i.p. It was Elevated Plus Maze Test (EPMT)
dissolved in 0.9% normal saline and sonicated for 10 min Behavioural symptoms were measured in the elevated
in ultrasonicated bath.[15] Ketamine was purchased from plus-maze 2 weeks after the final injection. The maze
Troikaa Pharmaceutical Ltd and was administered (30 was made of black polyvinyl chloride and had two open
mg/kg/day, i.p).[25] Risperidone (2 mg/kg/day, i.p) was and two closed arms (50×10×40 cm) mounted 50 cm
dissolved in 0.4 molL-1 tartaric acid.[29] above the floor. The floor of the arms was smooth. Light
levels were 30 or 400 lx. Bright illumination is
Behavioural symptoms were induced by intraperitoneal considered more stressful to animals. A rat was placed in
injection of Ket (30 mg/kg/day) for two weeks. Sub- the central platform of the apparatus facing a closed arm.
anaesthetic dose of ketamine produced behavioural This is expressed as the animal spending more time in
the enclosed arms. This model is based on rodent’s

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Iqbal et al. European Journal of Biomedical and Pharmaceutical Sciences

aversion of open spaces which involves avoidance of 2.4.7. SOD activity[34]

open areas by confining movements to enclose space.[31] The supernatant was assayed for SOD activity by
following the inhibition of pyrogallol autoxidation. 100
2.4.2. Locomotor Activity monitoring µl of cytosolic supernatant was added to Tris HCL buffer
It was observed in open field arenas consisting of an (pH 8.5). The final volume of 3 ml was adjusted with the
acrylic box (40.6×40.6×40.6 cm3) accommodated with same buffer. At least 25µl of pyrogallol was added and
two photo beam frames (16 beams/dimension; 2.5 cm change in absorbance at 420 nm was recorded at 1 min
between beams; Coulbourn Instruments, Allentown, PA). interval for 3 min. The increase in absorbance at 420 nm
The horizontal locomotor activity was recorded by the after the addition of pyrogallol was inhibited by the
lower frame (2.5 cm above the arena floor) while the presence of SOD.
upper frame (15 cm above the floor) records rearing. The
open field chamber was joined to a computer running 2.4.8. Estimation of brain AChE[35]
software (True Scan 2.0 version, Coulbourn Instrument, Rat brain was harvested by decapitation, immediately
Allentown, PA) that records beam break (100 ms after elevated plus maze test, weighed and kept at -70°C
sampling rate). Rats were kept for half an hour in home until AChE assay. The whole brain AChE activity was
cage for habituation. Then they were place in an open measured according to the method of Ellman et al., 1961.
field chamber for half an hour prior to observe the A known weight of the brain tissue was homogenized in
locomotor activity.[4] Locomotor activity was recorded 0.32 M sucrose solution to get a 10% homogenate that
for 20 minutes during which different parameters of was centrifuged at 3000 rpm for 15 minutes followed by
horizontal locomotion activity were recorded for each centrifugation at 10,000 rpm for 10 min at a constant
rat. temperature 4°C. 1 ml. of supernatant was mixed with 9
ml of sucrose solution to get a 1% post mitochondrial
2.4.4. Biochemical estimations supernatant (PMS). Test samples were prepared by
Brain was harvested by decapitation, immediately after mixing 2.7 ml of phosphate buffer, 0.1ml of DTNB and
behavioural studies, weighed and kept at -70°C till the 0.1 ml of PMS. Reaction mixture was taken in a cuvette
time it was used for assay of oxidative stress marker i.e. and pre-incubated for 5 min and 0.1 ml of acetyl
thiobarbituric acid reactive substances (TBARS), thiocholine iodide was added to the mixture to initiate
reduced glutathione (GSH), superoxide dismutase the reaction and immediately absorbance was recorded at
(SOD), were measured to establish antioxidant properties 412 nm for 3 min interval. Protein was determined
of Preg S and acetylcholine esterase activity (AChE) in according to well established method.[36]
rats brain.
2.5. Statistical Analysis
2.4.5. TBARS estimation[32] Data were expressed as the mean ± SEM. For a statistical
One ml of the suspension medium was taken from the analysis, group means was compared by one-way
supernatant of the 10% tissue homogenate and analysis of variance (ANOVA) followed by Tukey-
centrifuged at 10,000 rpm. 0.5 ml of 30% TCA followed Kramer multiple comparison tests which can be used to
by 0.5 ml of 0.8% TBA was added to it. The tubes were identify differences between groups. P value < 0.05 was
covered with aluminum foils and were kept in a shaking considered significant. Statistical analysis was carried
water bath for 30 min at 80оC. After 30 min, the tubes out using Graph Pad Prism 5.00.288 (Graph pad software
were taken out and were kept in ice-cold water for 10 San Diego, CA).
min. They were then centrifuged at 3000 rpm for 15 min.
The absorbance of the supernatant was read at 540 nm at 3.0. RESULTS
room temperature against an appropriate blank. Blank 3.1. Spontaneous Alternation Behaviour (SAB)
consisted of 1.0 ml distilled water, 0.5ml 30% TCA, There was slightly decrease in the percentage alternation
0.5ml 0.8% TBA. of animals in pregnenolone sulphate per se (11.86
mg/kg, Preg S) and risperidone (2 mg/kg, Risp) groups
2.4.6. GSH level[33] as compared to control group. However, a significant
A known weight of tissue ranging from (300-600 mg) increased in the percentage alternation was observed in
was homogenized in 5-8 ml of 0.02 M EDTA and then combination with ketamine (30 mg/kg, Ket) as compared
4.0 ml of cold distilled water was added to it. After to control group (p<0.05 (Fig 1).
mixing it well, 1 ml of 50% trichloroacetic acid (TCA)
was added and shaken intermittently for 10 min using a 3.2. Elevated plus maze test (EPMT)
vortex mixer. After 10 min the contents were transferred Administration of Preg S increased the time spent in
to centrifuge tube (rinsed in EDTA) and centrifuged at open arms and reduced the time spent in closed arms as
6000 rpm for 15 min. Following centrifugation, 2ml of compared to ket treated groups where closed arm entry
the supernatant was mixed with 4.0 ml of 0.4 M tris was highly significant (P<0.001) as compared to control
buffer (pH 8.9). The whole solution was mixed well and group. Total no. of open arm entries decreased in ket
0.1 ml of 0.01 M DTNB was added to it. Absorbance treated groups when compared with control groups and
was read within 5 min of the addition of DTNB at 412 Preg S per se groups showed highly significant results
nm against a reagent blank with no homogenate. (p< 0.001) (Table1).

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3.3. Locomotor activity monitoring study demonstrated relieving and improvement in the
Total movements, move time, move distance, mean stress and anxiety by Preg S which may be useful in
velocity were increased and rest time was decreased in ameliorating the symptoms of SCZ.
Ket treated group as compared with control group.
However combination of Preg S and Ris pre-treated SAB has been used as a measure of recording short term
groups showed highly significant reduction in total memory.[37] In the present study it was also observed that
movements, move time, move distance, mean velocity upon administration of Ket, the % alteration was
and increased in rest time as compared with Ket group increased as compared with saline treated groups in
(P<0.001) (Table 2). elevated plus maze- test. Combination of Preg S with Ket
reduced the % alternation suggesting its role in learning
3.4. Biochemical parameters and memory and anti-amnestic effect. These behavioural
3.4.1. GSH observations were corroborated with a decrease in AChE
Preg S (11.86 mg/kg) treatment showed significant activity in rat brain by Preg S thus, confirming its anti-
increase in GSH levels (p<0.05) and Ket administration amnesic activity against Ket induced amnesia in elevated
(30 mg/kg) resulted in decrease in GSH level however, plus maze model.
co-administration showed an increase in GSH level as
compared to their respective controls (Table 3). Total movement time, rest time and horizontal activity
were recorded in activity monitoring system to assess the
3.4.2. TBARS LA. Total movement time was increased in ketamine
Ket pre-treatment produced increased in TBARS value treated groups as compared with control groups. Preg S
as compared to control group (p<0.01). Preg S showed a combination groups showed a highly significant
reduction in TBARS level (p<0.05) as compared to reduction in LA as compared with ketamine group. N-
control group. Combination of Preg S and ketamine Methyl-D Aspartate (NMDA) receptor antagonists,
administration resulted in reduction of TBARS value phencyclidine and dizocilpine (MK-801), also produce
slightly more than control groups (Table 3). similar behavioural effects in rodents characterized by
increased LA similarly as our observations. In particular,
3.4.3. SOD activity NMDA antagonist-induced hyperlocomotion has been
SOD activity was significantly reduced in ketamine used to compare the effects of typical and atypical anti-
group (P< 0.001), however combination of Preg S and psychotic drugs in the NMDA-model of schizophrenic
Ket showed marked increased in SOD activity as symptoms.[38] Thus, Preg S may be a good candidate to
compared to control groups. (Table 3). have effects produced by NMDA receptor antagonistic
activity.[8] Recent studies provide additional evidence
3.4.4. AChE activity that Preg S may function as an endogenous
Ket (30 mg/kg) produced significant increased in acetyl neurotransmitter or neuromodulator, creating renewed
cholinesterase activity as compared to their control interest in the identification of novel neuroactive steroid
(p<0.05). Combination of Preg S and Ket treated animals targets for pharmacological intervention.[24,39]
showed decreased in AChE activity as compared to
control group (Fig. 2). Evidence for increased oxidative stress in chronic SCZ
patients is primarily based on the altered levels of
4.0. DISCUSSION antioxidants enzymes, free radical production or reactive
Intraperitoneal injection of multiple sub-anaesthetic dose oxygen species (ROS) can cause cellular damage or
of Ket (30 mg/kg) for two weeks as reported by earlier neuronal death, because oxidation of cellular components
reported study[25] demonstrated behavioural symptoms like lipid, protein and DNA and alteration of signalling
and memory impairment similar to that of schizophrenic pathway that finally promote the damage of cells.[40]
symptoms. The present study evaluated Preg S in
behavioural tests which include EPMT and SAB Multiple Sub-anaesthetic dose of Ket (30 mg/kg, i.p)
including the locomotor activity (LA) in rats. showed significant (p<0.01) increased in TBARS value
as compare to control treated groups. Impaired
During the EPMT, total time spent in closed arm, total antioxidant defences are suggested to participate in the
time spent in open arms, % preference to closed arm and pathophysiology of schizophrenia and other
open arm were recorded, animal spent more time in neurodegenerative conditions. Altered SOD and
closed arm as compared to open arm in group treated increased lipid peroxidation, measured by the TBARS,
with ketamine but when it was treated with Preg S, there are increased in schizophrenic patients.[41] Preg S showed
was an enhancement in the time spent in open arms as a highly significant reduction in TBARS level,
compared to closed arm which showed that rat suggesting its neuroprotective and/or anti-oxidant
demonstrated aversion toward open arm entries which properties.
indicates that anxiety and stress like symptoms were
reduced which is a core symptoms of schizophrenia. GSH represents main cellular non-protein antioxidant
Others symptoms also recorded in elevated plus maze and redox regulator in protecting nervous tissue against
like freezing to open space for long time. The present ROS[42] and in modulating redox sensitive sites,

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 Iqbal et al. European Journal of Biomedical and Pharmaceutical Sciences

including NMDA receptors. GSH level was significantly showed that neurosteroids play important role to
(p< 0.001) decreased in ketamine treated group as preventing oxidative damage cause neuronal cell death
compared to control group, after Preg S administration and apoptosis in schizophrenia by restoring this enzyme.
GSH level was increased, combination of Preg S and
ketamine show slightly increased in GSH level as AChE was increased in Ket treated rats (p<0.001) as
compared to Ket which also showed neuroprotective compared to control, Preg S showed marked decreased in
and/or anti-oxidant properties of drug. AChE activity. ACh release from rat hippocampus is
linked to cognitive function[43]s. Thus, indirectly our
SOD is an important enzyme in reducing oxidative study confirms that Preg S increased the Ach levels and
stress, in the present study there was a decrease in Ket improved the cognitive function.
treated rats, but highly increased in Preg S treated group

Table 1: Effect of Pregnenolone sulphate on Elevated Plus Maze test in rats.

Groups Drug Dosage % Time spent Total no. of Arm entries
(n=6) treatment (mg/kg) Open arm Close arm Open arm Close arm
I Normal Control 1 ml 65 ± 3.26 36.5 ± 2.56 14.89 ± 1.00 10.33 ± 0.88
II Ketamine 30 25.5 ± 2.65** 79.55 ±1.73*** 7.61 ± 0.40*** 16 ± 1.15**
## ## *##
III Preg S per se 11.86 66.75 ± 1.7 30.44 ± 0.96 12.36±0.49 14.66 ±0.88*#
### ## ###
IV Risp Per se 2 61.6 ± 2.7 35.00 ± 2.24 15.99±0.31 11.33±0.79##
V Preg S + ket 11.86 + 30 71.66 ± 1.39 32.81±1.31 9.97 ± 0.41 9.00 ± 0.97
VI Risp+ ket 2 + 30 66.69 ±1.88 40.30± 1.17 13.67 ± 0.67 8.66 ± 0.66
All the values were expressed as mean ±SEM and each data point was the average of 6 animals in each group (n=6).
Statistical analysis was carried out using analysis of variance (ANOVA) followed by Tukey-Kramer multiple
comparison test. P <0.05 was considered significant. * p<0.05, **p<0.01, *** p<0.001 when compared with control, #
p<0.05, ## p<0.01, ### p<0.001 when compared with ketamine. Preg S= Pregrenolone Sulphate; Risp =Risperidone;
ket=Ketamine

Table 2: Effects of Pregnenolone sulphate on ketamine induced locomotor activity in rats.

Average Mean Total
Groups Horizontal Move Rest time
treatment dist/move velocity movement
(n=6) activity(cm) time (s) (s)
(cm) (cm/s) (#)
Normal Control (1ml/kg, 2068.37 243± 682.57 2.58 2.73± 563.38
I
i.p) ±98.65 15.68 ±14.98 ±0.17 0.11 ±23.78
Ketamine 6082.62 ± 738 ± 246 5.16± 8.19 1689
II
(30 mg/kg, i.p) 252.70** 16.87** ±15.94** 0.35** ±0.07** ±78.26**
Preg S per se 1974.71 165 671± 1.89 ± 2.81 452.93
III
(11.86 mg/kg, i.p) ± 92.51**## ±8.79**## 8.57*# 0.06*## ±0.06# ±26.31*##
Risp per se 1052 232± 693.67 2.36 2.63 611.52
IV
(2 mg/kg, i.p) ±35.69**### 25.68## ±10.31*### ±0.04*## ±0.02### ±11.04**###
Preg S (11.86 mg/kg, i.p) 3032 ± 308 ± 612.65 ± 3.15± 3.18 ± 862.73
V
+ ket (30 mg/kg, i.p) 203.54 15.73 15.54 0.13 0.12 ± 27.54
Risp (2 mg/kg, i.p) + ket 4031± 302 ± 602.70 ± 3.24 ± 3.82 ± 903.21
VI
(30 mg/kg, i.p) 263.27 24.78 25.73 0.24 0.21 ± 22.13
All the values were expressed as mean ±SEM and each data point was the average of 6 animals in each groups (n=6).
Statistical analysis was carried out using analysis of variance (ANOVA) followed by Tukey-Kramer multiple
comparison test. P <0.05 was considered significant. * p<0.05, **p<0.01, when compared with control, # p<0.05, ##
p<0.01, ### p<0.001 when compared with ketamine. Preg S =Pregrenolone Sulphate; Risp =Risperidone;
ket=Ketamine

Table 3. Effects of Pregnenolone sulphate on GSH, TBARS and SOD in rats.

Groups GSH TBARS (nmol/mg of SOD (Unit/mg of
Drugs treatment
(n=6) (µg/mg of protein) protein) protein)
I Normal Control (1ml/kg, i.p) 14.25 ± 0.40 6.28 ± 0.16 171.8 ± 13.86
II Ketamine (30 mg/kg,i.p) 7.49 ± 0.38*** 7.52 ± 0.11** 76.74 ± 6.94***
*##
III Preg S per se (11.86 mg/kg, i.p) 11.26 ± 0.28 6.14 ± 0.25## 164.83 ± 16.16*##
###
IV Risp per se (2 mg/kg.i.p) 13.85 ± 0.27 4.05 ± 0.21***### 175.44 ± 12.83###
##
V Preg S (11.86 mg/kg,i.p) + ket (30 mg/kg, i.p) 9.6 ± 0.63 6.95 ± 0.14 127.61 ± 5.26**##
VI Risp (2 mg/kg, i.p) + ket (30 mg/kg, i.p) 10.1 ± 0.87 6.28 ± 0.23## 140.32 ± 8.17***###

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Iqbal et al. European Journal of Biomedical and Pharmaceutical Sciences

All values were expressed as mean ± SEM, analyzed by ANOVA followed by Tukey- Kramer multiple comparison
test. P values <0.05 was considered significant and P value <0.001 was considered extremely significant. N= 6 number
of animals in each group* p<0.05, **p<0.01, *** p<0.001 when compared with control, ## p<0.01, ### p<0.001 when
compared with ketamine. Preg S =Pregrenolone Sulphate; Risp =Risperidone; ket=Ketamine

CONCLUSION Figure 2: Effects of Pregnenolone Sulphate on acetyl

Ket (30 mg/kg i.p) administered for 14 days produced cholinesterase esterase activity in elevated plus maze
significant changes in behaviour resembling to that of test in rat.
negative symptoms as well as cognitive memory All the values were expressed as mean ± SEM and each
impairment seen in schizophrenia. Preg S treated rats data point was the average of 6 animals in each group
spent more time in open arm in EPMT, decreased (n=6). Statistical analysis was carried out using analysis
percentage alteration and improved LA as compared to of variance (ANOVA) followed by Tukey-Kramer
Ket treated rats. These observations indicated the multiple comparison test. P<0.05 was considered
improvement in learning and memory with Preg S. significant * p<0.05, **p<0.01, *** p<0.001 when
Oxidative stress was observed in Ket treated rats and compared with control, # p<0.05, ## p<0.01, ###
various antioxidant parameters were estimated. The p<0.001 when compared with ketamine. Preg S
levels of SOD and GSH were reduced and TBARs level =Pregrenolone Sulphate; Risp =Risperidone; Ket =
was elevated as compared to control groups. Upon Preg Ketamine
S administration, ameliorating effects was observed in
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