Post on 19-Dec-2021
transcript
www.wjpps.com Vol 4, Issue 12, 2015.
905
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
PHYTOCHEMICAL EVALUATION & PHARMACOLOGICAL
SCREENING OF ETHANOLIC BARK EXTRACT OF
DOLICHONDRONE FALCATA & ROOT EXTRACT OF
SCROPHULARIA HYPERICIFOLIA FOR
PSYCHOPHARMACOLOGICAL ACTIVITIES IN MICE.
Mehnoor Farheen*¹, Sumera Mahreen Khatoon², Asfia Kouser³
*¹M.Pharm (Ph.D.) Associate Professor, Department of Pharmacology, Shadan Women’s
College of Pharmacy, JNTU, Hyderabad, India
²,³Pharmacology, Shadan Women’s College of Pharmacy, JNTU, Hyderabad, India.
ABSTRACT
Psychopharmacology is concerned with drug effects that modify the
behaviour of animal organisms. This study assessed the antidepressant,
antianxiety, antipsychotic, antiepileptic, hypnotic and sedative
activities of ethanolic extract of Dolichondrone falcata and
Scrophularia hypericifolia in diseases induced mice. Extraction of the
ethanolic extract from bark of Dolichondrone falcata and roots of
Scrophularia hypericifolia was performed by maceration. 24 mice were
divided into six groups. Group I consisted of normal mice that were
given only sterile saline solution and served as control group. Group II
mice consisted of normal mice induced with disease either through
drug or physically. Group III, IV, V consisted of normal mice induced
with depression, anxiety, psychosis, epilepsy. Oral administration of
extract resulted in significant antidepressant, antianxiety,
antipsychotic, antiepileptic, hypnotic and sedative effect and muscle relaxant activity. Group
VI consisted of normal mice induced with depression, anxiety, psychosis, epilepsy and were
treated with standard drug. The effect produced by the extracts were closely similar to the
standard drugs. In conclusion, the present study indicates that the ethanolic extract of
Dolichondrone falcata and Scrophularia hyperictifolia appears to exhibit antidepressant,
antianxiety, antipsychotic, antiepileptic, hypnotic and sedative activity, muscle relaxant
activity in disease induced mice.
WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES
SJIF Impact Factor 5.210
Volume 4, Issue 12, 905-934 RReesseeaarrcchh Article ISSN 2278 – 4357
Article Received on
05 Oct 2015,
Revised on 29 Oct 2015,
Accepted on 22 Nov 2015
*Correspondence for
Author
Dr. Mehnoor Farheen
M.Pharm (Ph.D.) Associate
Professor, Department of
Pharmacology, Shadan
Women’s College of
Pharmacy, JNTU,
Hyderabad, India.
www.wjpps.com Vol 4, Issue 12, 2015.
906
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
KEYWORDS: elevated plus maze, forced swim test, haloperidol induced catalepsy, cerebral
edema, neuronal degeneration.
INTRODUCTION
In this study the extracts of parts of the plants Dolichondrone Falcata and Scrophularia
Hypericifolia was evaluated for their use in the management of diseases like anxiety,
depression, memory disorder and was evaluated for sedative and hypnotic property. From the
thorough study and investigation of the available literature of plants. Both the plants have
shown that they serve as an important sources of many therapeutically efficient chemicals
like flavonoids, organic acids like sugar and terpenes and various other constituents. The
ethanolic extract of bark of dolichondrone falcata and roots of scrophularia hypericifolia was
evaluated for various psychopharmacological disorders in mice. The disease was induced by
drug or through physical induction.
REVIEW OF DRUG UNDER STUDY
Scrophularia hypericifolia [1, 2]
Scrophularia hypericifolia belong to the genus Scrophularia, represented by over 300 species
worldwide in Saudi Arabia, about 5 species were described. Genus Scrophularia is rich in
iridoid glycosides, phenylpropanoids and flavonoids.
Dolichondrone falcata [3, 4]
Dolichondrone falcata is a small deciduous tree in the Bignoniaceae family. It
is endemic to India.
MATERIALS AND METHODS
Collection and preparation of extract
Dried bark extract of dolichondrone falcata and dried root extract of scrophularia
hypericifolia were collected. The plants were taxonomically identified and authenticated by
DR. K. MADHAVA CHETTY assistant professor of botany department of pharmacognosy,
Sri Venkateshwara University, Tirupati.
Preparation of Plant Extracts
The plant extract was prepared by maceration [5]
process. Standard screening test [6]
of the
extract was carried out for various plant constituents using standard procedures. Acute
www.wjpps.com Vol 4, Issue 12, 2015.
907
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
toxicity studies [7]
were performed for the extracts according to the acute toxic classic method
as per the method of Litchfield and Wilcoxon (1949).
Experimental animals
Mice (20 – 50 gms) of either sex housed in standard conditions of temperature (55+ 5%) or
light (12 hrs. light /dark cycles) were used. Animals were randomly selected for grouping. All
experiments were performed according to the forms of the ethical conditions. (CPSCSEA).
Histopathological studies [8]
The animals were euthanized using anesthetic ether and their brains were dissected out. The
isolated organ was sliced into 5mm pieces and fixed in neural formalin (10% solution) for at
least 3 days. This was followed by dehydration with isopropyl alcohol of increasing strength
(70%, 80%, and 90%) for 12 hours each and the final dehydration was done using absolute
alcohol with about three changes for 12 hours each. Clearing was done by using chloroform
with two changes for 15 to 20 minutes each. After clearing the object pieces were subjected
to paraffin infiltration in automatic tissue unit. The organ pieces were then dropped into
molten paraffin quickly and allowed to cool. The blocks were cut using microtome to get
sections of thickness of 5µ. The sections were applied with egg albumin then allowed to
remain in an oven at 600C for 1 hour. Paraffin melts and egg albumin denatures, thereby
fixing tissues to the slide. Staining with Eosin and hematoxylin stain.
Fig 1: isolated brain of mice
Elevated plus maze /elevated zero maze[9]
The Elevated plus-maze (EPM) was used to evaluate the anxiolytic effects of the plant
extract. Group I Mice received normal saline, group II received control drug , group III mice
www.wjpps.com Vol 4, Issue 12, 2015.
908
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
received control drug (levodopa) and bark extract of dolichondrone falcata (400mg/kg),
group IV mice received control drug (levodopa) and root extract of scrophularia hypericifolia
(200mg/kg), group V were received control drug and combination of both extracts
(dolichondrone falcata + scrophularia hypericifolia) and group VI were received control drug
and standard drug (diazepam 3mg/kg) and 60 min later, they were individually placed in the
center of the EPM for 5 min. The number of entries to the open and enclosed arms, and time
spent in the open arms, were recorded. Diazepam (3 mg/kg, i.p.) was used as positive control.
Fig 4: elevated plus maze test
Locomotor activity[10]
The locomotor activity can be easily studied with the help of actophotometer in a digital
actophotometer, continuous beam of light falls on the photoelectric cells. After 30 min of
administration of extracts and 1hr of i.p administration animals were individually placed in
actophotometer and the digital count as the number of line crossing by animal due to beam
interruption, were recorded for 10 minutes.
Fig 5: locomotor activity
Novelty-induced hypophagia[11]
Rodents encountering a desirable food in a novel environment will consume very limited
quantities after considerable investigation. Mice tend to avoid exploration of novel open
www.wjpps.com Vol 4, Issue 12, 2015.
909
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
environments, yet are motivated to approach and consume palatable food. This inhibition of
feeding behavior has been termed hypophagia and is robust in both rats and mice. Singly
housed mice are trained to consume the palatable food source by introducing it to them in
their home cage for 30 min daily over three consecutive days. Diluted condensed milk in
plastic serological pipettes (10 mL) with attached sippers and rubber stoppers are mounted to
the wire cage lid. On the fourth day mice are tested in the home cage condition. Record the
latency to the first lick and the total volume consumed in 5-min intervals across the 30-min
session. Note any mice that do not consume any condensed milk. They should be excluded
from further testing as they failed the training protocol. On day 5, position the pipette in the
wire lid of the novel cage and place the mouse into the novel cage environment. Record
latency and total volume consumed.
Forced swim test [12]
The FST was carried out on mice according to the method of Porsolt et al. mice are
individually forced to swim inside a vertical plexiglass cylinder (height 40 cm, diameter 18
cm, containing 15 cm of water at 25o C). Mice when placed first time in cylinder are initially
highly active, vigorously swimming in circles, trying to climb the wall or diving to the
bottom. After 2-3 min this activity begins to subside and gets interspersed with phases of
immobility or floating of increasing duration. After 5 -6 min immobility reaches a plateau
where the animals remain immobile for approximately 80% of time. After 15 min the animals
are removed and allowed to dry in heated (32o C) enclosure and returned to their home cages.
After 24 hours test drug and standard drugs are administered. Animals were forced to swim
for 6 min, and the time spent in immobility during the last 5 min of a 6 min observation
period was recorded manually by the competent observer.
Fig 6: forced swim test
www.wjpps.com Vol 4, Issue 12, 2015.
910
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Tail suspension test [13]
Tail suspension test is a facile means of screening the antidepressant drugs. The immobility
displayed by mice, when subjected to unavoidable and inescapable stress, reflects behavioral
despair, which in turn may reflect depressive disorders in human. Clinically effective
antidepressants significantly reduce the immobility that mice display following active and
successful attempts to escape when suspended by tail. The total duration of immobility was
quantified during a test period of 5min. Mice were considered immobile when they were
completely remain motionless for at least 30s.
Fig 7: tail suspension test
Test for motor coordination (Rota-rod test) in mice [14]
It is one of the classical methods introduced by dunham and miya in 1957 for the evaluation
of drugs interfering with the motor co-ordination activity by testing their ability to remain on
a revolving rod. The apparatus consist of a horizontal wooden rod or metal rod coated with
rubber with 3cm diameter attached to a motor with the speed set at 2 rotations per minute.
Only those mice, which demonstrate their ability to remain on the revolving rod for at least
60s, are chosen for the test.
Fig 8: motor coordination test
www.wjpps.com Vol 4, Issue 12, 2015.
911
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Pentobarbital-induced hypnotic test[15]
This test is indicative of CNS depressant activity of the test compound. Hypnotic, sedatives,
tranquilizers and even antidepressants have been shown to prolong the sleeping time after the
single dose of pentobarbital.
The loss of righting reflex is used as a criterion for the duration of sleep in this model. After
30 min of drug administration the animals are then placed on their back on thermo-
statistically warmed pad (37o C) and the duration of the loss of righting reflex, starting from
the time of pentobarbital injection is measured until the animal regains the righting reflex.
Mean value of duration of anesthesia in min are recorded in control and experiment group.
Fig 9: pentobarbital induced hypnotic test
Haloperidol induced catalepsy in mice[16]
Catalepsy in mice is induced by certain neuroleptic drugs due to their inhibitory effect on the
nigrostriatal dopamine system. It has been defined as a failure to correct an externally
imposed nasal posture over a prolonged period of time. After 30 minutes of drug
administration they are placed individually into a translucent plastic boxes with a wooden
dowel mounted horizontally 10 cm from the floor and 4cm from the end of the box, the floor
of the box is covered with approximately 2cm of the bedding material. The animals are
allowed to adapt to the box for 5min. thereafter each animal is grasped gently around the
shoulders and under the forepaws and placed carefully on the dowel. The amount of time
spent with at least one forepaw on the bar is determined. When the animal removes its paws,
the time is recorded and the mice is repositioned on the bar. Four trials are conducted for each
animal at 30, 60, 90, 120, an animal is considered cataleptic if it remains on the bar for 60
www.wjpps.com Vol 4, Issue 12, 2015.
912
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
sec. the percentage of cataleptic animals is calculated. The phenomenon of catalepsy predicts
antipsychotic activity.
Fig 10: haloperidol induced catalepsy
Step-down inhibitory avoidance[17]
Rodents when placed in an open field, are known to spend most of the time close to walls or
corners. When they are placed on an elevated platform in the center of rectangular
compartments, they instantaneously step down to the floor and explore the enclosure to
approach the wall or corner. The inhibitory avoidance model (IAM) was used to assess the
effects of the plant extract on the animals’ memory. In the training session (day 1),
immediately after stepping down and placing their paws on the grid, the animals received a
0.3 mA 2 s scrambled foot shock and were immediately withdrawn from the cage. Drugs
were given immediately after the training session to investigate their effect on memory
consolidation. Twenty-four hours later, in the test sessions (day 2), no foot shock was given
and the stepdown latency was used as a measure of retention (to a ceiling of 300 s).
Hole cross test[18]
The method was adopted as described by Takagi el al. (1971). A wooden partition was fixed
in the middle of a cage having a size of 30 x 20 x 14 cm. a hole or 3 cm diameter was made at
a height of 7.5 cm in the center of the cage. Diazepam (1mg/kg) is used as a standard drug.
After 30, 60, 90 min of administration of drugs, the mice were placed in the apparatus and the
number of passage of mouse through the hole from one chamber to other was counted for a
period of 3 min.
www.wjpps.com Vol 4, Issue 12, 2015.
913
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Fig 11: hole cross test
Head dip test[19]
Boisser et al (1964) have used an open field with holes on the bottom into which the animals
cold poke their noses. The phenomenon indicates the curiosity component of the behavior of
mice. Swiss albino mice of either sex are used. The hole board has a size of 40 E and 40 cm
in which 16 holes of 3cm diameter each are evenly distributed on the floor. Nose pocking is
considered as an indication of curiosity/ exploratory behaviour and is measured by visual
observation or by electronic devices in more recent modifications. 30 minutes after
administration the animals are subjected to test for 5min. Diazepam is used as standard drug.
The average count of nose poking of treated animals is calculated as the percentage of control
animals.
Fig 12: head dip test
Isoniazid – induced convulsion[20]
Isoniazid is known to produce clonic – tonic seizures which can be used as one of the battery
of tests for anticonvulsant activity. Thirty minutes after i.p and 1 hour after oral
administration the animals are subjected to subcutaneous administration of isoniazid. During
www.wjpps.com Vol 4, Issue 12, 2015.
914
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
the next 2 hours the occurrence of colonic and tonic seizures and death is recorded. The
percentage of seizures and death occurring in control mice is taken as 100%. The suppression
of these effects in the treated group or the protection against death is calculated as percentage
of controls.
Fig 13: isoniazid induced convulsions
Effect of plant on normothermic animal[21]
Antidepressants have been shown to prevent the hypothermia induced by apomorphine in
mice. Compounds with a significant noradrenergic or dopaminergic activity are active against
apomorphine induced hypothermia but not the antidepressants which act mainly through
serotoninergic mechanism. A time vs temperature curve is constructed by mean temperature
of each group against time. The area under the curve (AUC) is calculated for all groups and
converted into percent inhibition of apomorphine induced hypothermia in.
Fig 14: effect of plant extract on normothermic animal control group.
www.wjpps.com Vol 4, Issue 12, 2015.
915
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Antipsychotic induced weight gain [22]
The difference in the mice weight before the administration of drugs and after 0, 2, and 4
weeks of administrations of drugs was then compared.
Caffeine induced stereotypy and locomotion[23]
The method described by Randrup and Mnkvad (1967) was used for the stereotype
behavioural studies. Adult mice were randomized into five groups of 4 mice each. Group I
mice were received normal saline, group II mice were received caffeine as a control, group
III mice were given ethanolic bark extract of dolichonrone falcata (400mg/kg), group IV mice
were treated with ethanolic root extract of scrophularia hypericifolia (200mg/kg), group V
mice were treated with combination of both the extracts (DF+SH) and Group VI received
standard drug risperidone (1ml/kg) respectively. One hour after administration of saline and
extract and thirty minutes after administration of risperidone, group III, IV, V and VI mice
were given 2mg caffeine/kg. The signs of stereotype behaviour that included circling,
jumping, sniffing, and general locomotion were recorded for period of 2h using a hand held
tally counter (Irwin, 1968).
Stereotyped behaviour was scored as follows.
A. complete absence of stereotyped behaviour =0,
B. Presence of stereotyped movements of the head and intermittent sniffing = 1,
C. sniffing and chewing = 2,
D. chewing and intense licking = 3.
Evasion test [24]
The method of Turner (1965) was used, the animals were introduced into this rectangular box
with an inclined plane by which the mice can escape from the box, and the mice that escaped
within 5min from the rectangular box were selected for this test, 15 minutes after
administration of normal saline, diazepam as control, dolichondrone falcata and scrophularia
hypericifolia the animals (n=8) were placed in the box again and the number of mice
remaining in the box after 15 min in each group was noted.
www.wjpps.com Vol 4, Issue 12, 2015.
916
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Fig 15: evasion test
RESULTS
phytochemical analysis of ethanolic bark extract of dolichondrone falcata
TABLE 1: Phytochemical analysis of ethanolic bark extract of dolichondrone falcata.
Types of constituents Ethanolic extracts
Alkaloids -
Flavonoids +
Steroids +
Phenolic compounds +
Carbohydrates +
phytochemical analysis of ethanolic root extract of scrophularia hypericifolia
TABLE 2: Phytochemical analysis of ethanolic root extract of scrophularia
hypericifolia.
Types of constituents Ethanolic extracts
Alkaloids +
Flavonoids +
Iridoid glycosides +
Phenylpropanoids +
Carbohydrates +
Steroid -
www.wjpps.com Vol 4, Issue 12, 2015.
917
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Elevated plus maze.
TABLE 3: Effect of ethanolic extract of bark of dolichondrone falcata (test drug 1) and
root of scrophularia hypericifolia (test drug 2) and combination of both extracts (test
drug 3) and standard drug and standard drug on elevated plus maze test.
Group Drug treatment
Number of
entries in open
arms
Time spent in
open arm
Number of
entries in
closed arms
Time spent in
closed arms
Group I Normal saline (1ml) 2.750 ± 0.8539 1.500 ± 0.2887 3.500 ± 0.2887 1.000 ± 0.0
Group II Dolichondrone falcate
(DF)(400 mg/kg) 4.500 ± 0.5000 4.500 ± 0.2887 3.250 ± 0.4787 4.500 ±0.2887
Group III
Scrophularia
hypericifolia(SH)
(200mg/kg)
3.250 ± 0.2500 3.250 ± 0.2500 2.250 ± 0.2500 3.250 ± 0.4787
Group IV DF(200mg/kg) +
SH(100mg/kg) 3.500 ± 0.2887 3.500 ± 0.2887 2.250 ± 0.2500 3.500 ± 0.2887
Group V Diazepam(0.7mg/kg) 2.500 ± 0.2887 3.500 ± 0.2887 1.500 ± 0.2867 3.000 ± 0.4082
Data are presented as mean values (± S.E.M.). *P values: < 0.001 compared with treated
control. Statistical test employed was ANOVA.
Graph 1: open arm entries Graph 2: time spent in open arm
Graph 3: closed arm entries Graph 4: time spent in closed arm
www.wjpps.com Vol 4, Issue 12, 2015.
918
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Locomotor activity
TABLE 4: Effect of ethanolic extract of bark of dolichondrone Falcata (test drug 1) and
root of scrophularia hypericifolia (test drug 2) and combination of both extracts (test
drug 3) and standard drug on locomotor activity.
Group Drug treatment
Locomotor activity in 10 min Percentage
decrease in
activity Before
Treatment
After
Treatment
Group I Normal saline (1ml) 131.3 ± 5.154 138.3 ± 4.049 5%
Group II Caffeine (2mg/kg) 134.0 ± 3.719 138.8 ± 4.270 67.95%
Group III
Caffeine (2mg/kg) +
Dolichondrone falcata(DF)
(400mg/kg)
134.8 ± 2.287 58.75 ± 10.36 84%
Group IV
Caffeine (2mg/kg) +
Scrophularia
hypericifolia(SH) (200mg/kg)
1105 ± 7.089 46.50 ± 17.95 90.1%
Group V Caffeine (2mg/kg) + (DF
200mg/kg +SH 100mg/kg) 118.3 ± 3.065 22.75 ± 5.851 90.5%
Group VI Caffeine (2mg/kg) +
Diazepam(3mg/kg) 110.3 ± 3.705 24.25 ± 2.780 62%
(Observation period: 10 min) values are expressed as mean values ± S.E.M from 4 mice
significant at * *P < 0.01 as compared to control using one way ANOVA followed by
Dennett’s t- test.
Graph 5: locomotor activity score
www.wjpps.com Vol 4, Issue 12, 2015.
919
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Novelty induced hypophagia
TABLE 5: Effect of ethanolic extract of bark of dolichondrone Falcata (test drug 1) and
root of scrophularia hypericifolia (test drug 2) and combination of both extracts (test
drug 3) and standard drug on novelty induced hypophagia.
Group Drug Dose/treatment Latency to drink (sec)
I Normal saline 1ml 321.8 ± 10.48
II Dolichondrone falcate(DF) 400mg/kg 207.3 ± 9.961
III Scrophularia hypericifolia(SH) 200mg/kg 243.8 ± 16.75
IV DF+SH 200mg/kg+ 100mg/kg 255.0 ±9.574
V Imipramine 10mg/kg 231.0 ± 5.260
Data are presented as mean values (± S.E.M.). *P < 0.05 compared with vehicle‐treated
control with ANOVA.
GRAPH 6: Novelty induced hypophagia test
Forced swim test
TABLE 6: Effect of ethanolic extract of bark of dolichondrone Falcata (test drug 1) and
root of scrophularia hypericifolia (test drug 2) and combination of both extracts (test
drug 3) and standard drug on forced swim test.
Group Drug Dose (mg/kg) Immobility (sec)
I Normal saline 1ml 137.5 ±1.190
II Dolichondrone falcate(DF) 400mg/kg 110.8 ± 11.80
III Scrophularia hypericifolia(SH) 200mg/kg 115.5 ± 3.227
IV DF + SH 200mg/kg + 100mg/kg 100.8 ± 4.732
V Imipramine 10mg/kg 95.3 ± 2.394
Data are presented as mean values (± S.E.M.). *P < 0.05 compared with vehicle‐treated
control.
www.wjpps.com Vol 4, Issue 12, 2015.
920
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
GRAPH 7: forced swim test
Tail suspension test
TABLE 7: effect of ethanolic extract of bark of dolichondrone Falcata (test drug 1) and
root of scrophularia hypericifolia (test drug 2) and combination of both extracts (test
drug 3) and standard drug on test.
Group Drug Dose Immobility(sec)
I Normal saline 1ml 173.3 ± 1.181
II Dolichondrone falcate(DF) 400mg/kg 95.75 ± 0.7500
III Scrophularia hypericifolia(SH) 200mg/kg 97.75 ± 0.4787
IV DF + SH 200mg/kg + 100mg/kg 97.50 ± 0.5000
V Imipramine 10mg/kg 65.75 ± 2.136
Data are presented as mean values (± S.E.M.). *P < 0.05 compared with vehicle‐treated
control.
GRAPH 8: Tail suspension test
www.wjpps.com Vol 4, Issue 12, 2015.
921
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Rota rod test
TABLE 8: Effect of ethanolic extract of bark of dolichondrone Falcata (test drug 1) and
root of scrophularia hypericifolia (test drug 2) and combination of both extracts (test
drug 3) and standard drug on Rota rod test.
Group Dose Time spend on rod in sec
0 sec 30 sec 60 sec 120 sec
I Normal saline (1ml) 262.5 ± 2.255 266.5 ± 5.606 262.5 ± 4.211 280.5 ± 6.357
II Dolichondrone falcate(DF)
(400mg/kg) 255.0 ± 2.887 137.5 ± 8.780 117.5 ± 11.09 130.0 ± 7.071
III Scrophularia hypericifolia(SH)
(200mg/kg 246.3 ± 8.985 125.0 ± 11.37 123.8 ± 25.44 132.5 ± 11.81
IV DF (200mg/kg)+SH
(100mg/kg) 259.0 ±5.115 128.8 ± 7.181 120.0 ± 4.564 123.8 ± 3.750
V Diazepam (5mg/kg) 263.8 ± 2.394 51.50 ± 2.901 43.50 ± 1.936 59.75 ± 4.090
Data are presented as mean values (± S.E.M.). *P < 0.05 compared with vehicle‐treated
control.
Graph 9: Rota rod test
Pentobarbital induced sleeping time
TABLE 9: Effect of ethanolic extract of bark of dolichondrone Falcata (test drug 1) and
root of scrophularia hypericifolia (test drug 2) and combination of both extracts (test
drug 3) and standard drug on pentobarbtone induced sleeping time.
Group Drug Dose/treatment mg/kg Onset of
sleep(min)
Duration of
sleep(min)
I Normal saline - - -
II Pentobarbital 30mg/kg 3.9 ± 0.33 26.9 ± 2.3
III Pentobarbital +Dolichondrone
falcate (DF) 30mg/kg + 400mg/kg 3.2 ± 0.24 41.1 ± 4.4
IV Pentobarbital + Scrophularia
hypericifolia (SH) 30mg/kg + 200mg/kg 5.2 ± 0.56 53.3 ± 6.5
V Pentobarbital + DF + SH 30mg/kg + 4.3 ± 0.15 80.3 ± 4.6
www.wjpps.com Vol 4, Issue 12, 2015.
922
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
200mg/kg+100mg/kg
VI Pentobarbital 30mg/kg +
Diazepam(5mg/kg) 30mg/kg + 4mg/kg 2.5 ± 0.16 100.7 ± 2.94
Data are presented as mean values (± S.E.M.). *P < 0.001 compared with vehicle‐treated
control.
Graph 10: Pentobarbital induced sleep time
Haloperidol induced catalepsy in mice
TABLE 10: Effect of ethanolic extract of bark of dolichondrone Falcata (test drug 1)
and root of scrophularia hypericifolia (test drug 2) and combination of both extracts
(test drug 3) and standard drug on haloperidol induced catalepsy.
Group Drug Mean cataleptic score(sec)
30 min 60 min 90 min 120 min
I NS (1ml) 25.50 ± 0.2887 51.50 ± 0.2887 115.0 ± 2.887 100.0 ± 0.0
II Haloperidol (1mg/kg) 26.25 ± 2.394 23.75 ± 9.437 33.75 ± 3.750 35.00 ± 4.564
III Haloperidol + Dolichondrone
falcate(DF) (400mg/kg) 5.500 ± 0.2887 3.500 ± 0.2887 3.500 ± 0.2887 7.250±0.4787
IV Haloperidol+Scrophularia
hypericifolia(SH) (200mg/kg) 11.00 ± 0.4082 5.000 ± 0.4082 14.75 ± 0.2500 13.50 ±0.866
V Haloperidol + DF (200mg/kg) +
SH (100mg/kg) 14.50 ± 0.2887 11.75 ± 0.2500 13.25 ± 0.4787 9.500±0.2887
VI Haloperidol + Risperidone
(1mg/kg) 6.750 ±0.2500 4.750 ± 0.4787 6.250 ± 0.2500 4.750±0.4787
Data are presented as mean values (± S.E.M.). *P < 0.001 compared with vehicle‐treated
control.
www.wjpps.com Vol 4, Issue 12, 2015.
923
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Graph 11: Haloperidol induced catalepsy
Step down inhibitory avoidance
TABLE 11: Effect of ethanolic extract of bark of dolichondrone Falcata (test drug 1)
and root of scrophularia hypericifolia (test drug 2) and combination of both extracts
(test drug 3) and standard drug on step down inhibitory avoidance test.
Treatment Dose mg/kg Latency /time to step down in sec
I Normal saline(1ml) 193.8 ± 10.68
II Dolichondrone falcate (DF)400mg/kg 475.0 ± 20.62
III Scrophularia hypericifolia (SH)200mg/kg 473.8 ± 22.67
IV DF 200mg/kg+ SH 100mg/kg 307.5 ± 12.99
V Paracetamol (200mg/kg) 502.5 ±13.15
Data are presented as mean values (± S.E.M.). *P < 0.001 compared with vehicle‐treated
control.
Graph 12: step down test
www.wjpps.com Vol 4, Issue 12, 2015.
924
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Head dip test
TABLE 12: Effect of ethanolic extract of bark of dolichondrone Falcata (test drug 1)
and root of scrophularia hypericifolia (test drug 2) and combination of both extracts
(test drug 3) and standard drug on head dip test.
Group Drug Dose No. of head
dipping
I Normal saline 1ml 39.25 ± 0.4787
II Dolichondrone falcate (DF) 400mg/kg 22.25 ± 0.478
III Scrophularia hypericifolia
(SH) 200mg/kg 26.75 ± 0.2500
IV DF + SH 200mg/kg + 100 mg/kg 19.25 ± 0.4787
V Diazepam 2mg/kg 21.25 ± 0.4787
Data are presented as mean values (± S.E.M.). *P < 0.001 compared with vehicle‐treated
control.
Graph 13: head dip test
Hole cross test
TABLE 13: Effect of ethanolic extract of bark of dolichondrone Falcata (test drug 1)
and root of scrophularia hypericifolia (test drug 2) and combination of both extracts
(test drug 3) and standard drug on hole cross test.
Group Drug /Dose No. of movements (sec)
30 min 60 min 90 min
I Normal saline (1ml) 7.750 ± 0.2500 7.750 ± 0.2500 8.750 ± 0.2500
II Dolichondrone falcata (DF)
(400mg/kg) 1.500 ± 0.2887 2.000 ± 0.0 3.500 ± 0.5000
III Scrophularia hypericifolia
(SH) (200mg/kg) 4.000 ± 0.4082 3.000 ± 0.4082 4.000 ± 0.4082
IV DF (200mg/kg) + SH
(100mg/kg) 6.250 ± 0.4787 2.250 ± 0.4787 2.250 ± 0.4787
V Diazepam (1mg/kg) 1.250 ± 0.2500 6.250 ± 0.2500 4.250 ± 0.2500
Data are presented as mean values (± S.E.M.). *P < 0.001 compared with vehicle treated
control.
www.wjpps.com Vol 4, Issue 12, 2015.
925
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Graph 14: hole cross test
Isoniazid induced convulsions
TABLE 14: Effect of ethanolic extract of bark of dolichondrone Falcata (test drug 1)
and root of scrophularia hypericifolia (test drug 2) and combination of both extracts
(test drug 3) and standard drug on isoniazid induced convulsions.
Group Drug Dose/treatment
mg/kg
Onset of
seizure
%
mortality
%
protection
I Normal saline - - - -
II Isoniazid 300mg/kg 81.2 ± 4.21 100 0
III Isoniazid+Dolichondrone
falcate (DF)
300 mg/kg+
400mg/kg 145.2 ± 6.2 0 100
IV Isoniazid +Scrophularia
hypericifolia (SH) 300mg/kg+200mg/kg 133.0 ± 6.38 25 75
V Isoniazid +DF + SH 300mg/kg+200mg/kg
+ 100 mg/kg 159.0 ± 6.38 0 100
VI Isoniazid+Diazepam 300mg/kg +2mg/kg A 0 100
Data are presented as mean values (± S.E.M.). *P < 0.001 compared with vehicle treated
control.
Graph 15: isoniazid induced convulsions
www.wjpps.com Vol 4, Issue 12, 2015.
926
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Effect of drug on normothermic animals.
TABLE 15: Effect of ethanolic extract of bark of dolichondrone falcata (test drug 1)
and root of scrophularia hypericifolia (test drug 2) and combination of both extracts
(test drug 3) and standard drug on normothermic animals (mice).
Group Drug Dose/treatment
mg/kg
After drug administration
30min 60min
I Normal saline 1ml 36.55 ± 0.1100 36.66 ± 0.0
II Dolichondrone
falcate(DF) 400mg/kg 35.09 ± 0.5657 34.61 ± 0.1330
III Scrophularia
hypericifolia (SH) 200mg/kg 34.86 ± 0.1400 35.08 ± 0.05266
IV DF + SH 200mg+ 100mg 35.29 ± 0.1961 35.36 ± 0.1324
Data are presented as mean values (± S.E.M.). *P < 0.001 compared with vehicle treated
control.
Graph 16: effect of drug on normothermic animal
Antipsychotic induced weight gain.
Table 16: Effect of ethanolic extract of bark of dolichondrone Falcata (test drug 1) and
root of scrophularia hypericifolia (test drug 2) and combination of both extracts (test
drug 3) and standard drug on antipsychotic induced weight gain in mice.
Group Drug / dose (mg/kg) Weight gain
0 weeks 2 weeks 4 weeks
I Normal saline(1ml) 23.75 ± 2.394 25.00 ± 0.0 24.75 ± 2.056
II Risperidone (1mg/kg) 24.00 ± 1.354 35.25 ± 2.056 39.50 ± 2.021
III Risperidone +dolichondrone
falcate (400mg/kg) 27.75 ± 1.315 37.00 ± 1.915 42.50 ± 1.443
IV Risperidone + scrophularia
hypericifolia (200mg/kg) 23.75 ± 1.250 36.75 ±1.974 45.50 ±1.658
V Risperidone + (DF200mg/kg
+ SH100mg/kg) 22.50 ± 1.443 36.75 ± 1.974 46.00 ± 1.354
Data are presented as mean values (± S.E.M.). *P < 0.001 compared with vehicle‐treated
control.
www.wjpps.com Vol 4, Issue 12, 2015.
927
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Graph 17: weight gain
Caffeine induced stereotypy
TABLE 17: Effect of ethanolic extract of bark of dolichondrone Falcata (test drug 1)
and root of scrophularia hypericifolia (test drug 2) and combination of both extracts
(test drug 3) and standard drug on caffeine induced stereotypy.
Treatment Dose mg/kg Score for stereotypy
behavior
Control (caffeine) 2mg/kg 2.72 ± 0.09
Caffeine + Dolichondrone falcate 2mg/kg + 400 mg/kg 1.34 ± 0.05
Caffeine + scrophularia hypericifolia
(200mg/kg) 2mg/kg + 200mg/kg 1.22 ± 0.04
Caffeine + (DF 200mg/kg + SH
100mg/kg)
2mg/kg + 200mg/kg
+ 100mg/kg 0.66 ± 0.04
Caffeine + Haloperidol 2mg/kg + 1mg/kg 0.00 ± 0.0
Data are presented as mean values (± S.E.M.). *P < 0.001 compared with vehicle treated
control.
Graph 18: caffeine induced stereotypy
www.wjpps.com Vol 4, Issue 12, 2015.
928
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Evasion test
TABLE 18: Effect of ethanolic extract of bark of dolichondrone Falcata (test drug 1)
and root of scrophularia hypericifolia (test drug 2) and combination of both extracts
(test drug 3) and standard drug on evasion test in mice.
Treatment Dose No. of mice remaining
in box after 5 min
Normal saline 1ml 0
Dolichondrone falcata (DF) 400mg/kg 3
Scrophularia hypericifolia (SH) 200mg/kg 2
DF + SH 200mg/kg +100mg/kg 6
Diazepam 2mg/kg 6
Data are presented as mean values (± S.E.M.). *P < 0.001 compared with vehicle treated
control.
Graph 19: evasion test
Histopathological Reports.
Fig 16: microphotograph of brain of mouse Fig 17: microphotograph of brain of mouse
treated with normal saline for one month with Ethanolic Bark Extract of
Dolichondrone falcata. one month.
www.wjpps.com Vol 4, Issue 12, 2015.
929
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Fig 18: microphotograph of brain of
mouse treated with Ethanolic Root
Extract of scrophularia hypericifolia for
one month.
Fig 19: microphotograph of brain of
mouse treated with combination of
ethanolic bark extract of dolichondrone
falcata and root extract of scrophularia
hypericifolia.
Fig 20: microphotograph of brain of mouse treated with control group for one month.
DISCUSSION
The ethanolic extract of dolichondrone falcata and scrophularia hypericifolia gave positive
chemical reactions for glycosides, saponins, proteins, amino acids and flavonoids etc. No
toxic symptoms were observed for the drug up to of 400mg/kg body weight and 200mg/kg.
The extracts showed similar results when compared to standard drug diazepam. The
experimental result indicates that the extracts influence general behavior profile as evidenced
in awareness, alertness touch response, pinna reflex, righting reflex. However standard
www.wjpps.com Vol 4, Issue 12, 2015.
930
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
diazepam showed significant depression of all these responses. In Elevated plus maze in mice
Diazepam significantly increased the number of entries and time spent in open arms of
elevated plus maze. Dolichondrone falcata 400mg/kg and schrophularia hypericifolia
200mg/kg significantly increased the number of entries and percentage of time spent in open
arms as compared to vehicle (0.01% ethanol) treated mice. Diazepam and plant extracts
showed decrease in closed arm entries and time spent in closed arm. Therefore showed
significant antianxiety effect as compared to vehicle (0.01% ethanol) treated mice. However
scrophularia hypericifolia showed less result as compared to dolichondrone falcata.
Locomotor activity in mice showed that, both extracts significantly depressed the locomotor
activity at the tested dose level in mice. The activity was found to be maximum for
dolichondrone falcata and minimum for scrophularia hypericifolia. Further the activity of
extracts were more when compared with caffeine, a CNS depressant agent. In Novelty
induced hypophagia test on mice the latency to drink the condensed milk was decreased as
compared to normal saline. Both the extracts showed near similar results as that of standard
imipramine. In Forced swim test on mice drugs reliably decrease the duration of immobility
in animals during these tests. This decrease in duration of immobility is considered to have a
good predictive value in the evaluation of potential antidepressant agents (Porsolt RD,
et.al.1977). It was observed that the extract of Dolichondrone Falcata was more significant
than extract of Scrophularia Hypericifolia. In Tail suspension test on mice drugs reliably
decrease the duration of immobility in animals. This decrease in duration of immobility is
considered to have a good predictive value in the evaluation of potential antidepressant
agents (Porsolt RD, et.al.1977). It was observed that the extract of dolichondrone falcata was
more significant than extract of scrophularia hypericifolia.
The mean time duration on rotating rod was significantly decreased (P<0.001) and the
number of times animals falling down were increased in animals treated with Dolichondrone
falcata (400mg/kg) and Scrophularia hypericifolia (200mg/kg) as compared to control group,
indicating muscle relaxant activity and decline in motor coordination dolichondrone falcata
showed better result compared to scrophularia hypericifolia. However the extracts showed
good results in combination. In Pentobarbital induced sleeping test on mice, both extracts
increased sleep duration significantly modify the sleep latency. In agreement with the
previously published reports and as expected, diazepam significantly prolonged
pentobarbital-induced sleeping time, indicating that our study procedures were optimized.
www.wjpps.com Vol 4, Issue 12, 2015.
931
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Both the extracts has protective effects against haloperidol induced catalepsy, which is
comparable to standard drug. In Step down inhibitory avoidance on mice the extracts showed
enhanced memory and learning behavior in mice which is showed by delay in the stepping
down of mice. Both the drugs increased the number of head pokes significantly (P < 0.01) as
compared to the reference standard (diazepam, 1 mg/kg, i.p.)-treated group showed
significant increase in exploratory activity. In the present study, the effect of ethanol extract
of dolichondrone falcata and scrophularia hypericifolia on CNS has been evaluated by hole
cross test. The result indicated that the extract significantly decreased locomotor activity
which indicates it has CNS depressant activity. Locomotor activity refers to an increase in
alertness and decrease in locomotor activity considered as sedative effect. The major
inhibitory neurotransmitter in the central nervous system is Gamma-amino-butyric acid
(GABA). Different types of anxiolytic, muscle relaxant, sedative-hypnotic drugs are shown
their action through GABAA, that’s why the extracts may acts by membrane
hyperpolarization which potentiating GABA-ergic inhibition in the CNS that leads to either
decrease in the firing rate of critical neurons in the brain or direct activation of GABA
receptor by the extracts
Most of conventional antiepileptic drugs are associated with many side effects such as
neurotoxic effects, cognitive deficits and teratogenic effects which decrease their clinical
utility (Trimble, 1987; Yerby, 1988; Meador et al., 1990). Recently, the search for novel
pharmacotherapy from medicinal plants for neurological and psychiatric diseases has
progressed significantly owing to their less side effects and better tolerability (Zhang, 2004.
Isoniazid exerts its convulsive effect by inhibiting GABA synthesis (Costa et al., 1975). It is
a potent monoamine oxidase (MAO) inhibitor and a glutamic acid decarboxylase (GAD)
inhibitor (enzyme involved in GABA synthesis) thus increases the brain monoamine content
and inhibited GABA synthesis respectively thereby leading to CNS excitation and
convulsions (Wood and Peesker, 1973; Marcus and Coulsto, 1985). Isoniazid induced
seizures was carried out to further confirm the GABA enhancing activity of the plant extract.
The extracts when compared to control treated produced significant increase in the time of
onset of clonic seizures, this shows dose-dependent increase in the anticonvulsant activity.
In present study the effect of the extracts on body temperature of mice were evaluated.
When compared with the normal saline the extracts showed a slight decrease in the
temperature, there was no change with the normal saline administration after 60 minutes.
www.wjpps.com Vol 4, Issue 12, 2015.
932
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
Effect of ethanolic bark extract of dolichondrone falcata and ethanolic root extract of
scrophularia hypericifolia on Caffeine induced stereotypic behavior on mice. In present
study, we investigated effects of extract of dolichondrone falcata and scrophularia
hypericifolia against caffeine induced stereotypy. Caffeine directly activates dopamine
receptors in the brain and doses of the drug induced stereotyped behaviour (sniffing, licking
and gnawing). The stimulant effect of high doses of APM is attributed to activation of post
synaptic receptors in the CNS. The behavioral responses observed in animals after
administration of the dopamine agonist, caffeine are attributed to activation of D1 and D2
receptors. Stereotyped behaviour is more closely associated with the caudate striatum area of
brain. Treatment of alcoholic extracts of bark and roots of dolichondrone falcata and
scrophularia hypericifolia extracts antagonized spontaneous motor activity and also caffeine
induced hyperactivity in mice. Findings indicate that the appetite stimulation–weight gain
associated with AAPDs is mediated by activation of hypothalamic AMPK linked to blockade
of the histamine H1R. AMPK stimulation parallels the orexigenic actions of the drugs, with
clozapine and olanzapine producing the most marked effects. The drug actions are very
potent, with substantial effects evident at 5 nm concentration. They are selective, with effects
restricted largely to the arcuate and paraventricular nuclei of the hypothalamus. Orexigenic
potencies of AAPDs parallel their affinities for histamine H1Rs, These findings are in accord
with studies implicating central histamine in weight control as well as the orexigenic role of
the paraventricular and arcuate nuclei. Moreover, mice, like humans, manifest weight gain in
response to AAPDs, although inhibition of locomotor activity sometimes impairs
characterization of orexigenic actions. The ethanolic extracts showed significant weight gain
in mice thus showing the property of antipsychotic drugs.
The extracts produced a significant decrease in exploratory behaviour pattern as evident from
the results of evasion tests. Positive control diazepam inhibited total residual curiosity. The
histopathological report shows the presence of cerebral edema, mild neuronal degeneration
when treated with the ethanolic plant extracts for one month, these abnormalities were due to
the induction of disease in mice. However with the control drug there was a glial reaction
which is generally considered to be consequence of neuronal death in neuro degenerative
disease such as Alzheimer’s, Huntington and Parkinson disease. The effect of plant ethanolic
extracts on mice brain showed absence of glial reaction which may be due to the inhibition of
glial reaction by plant extracts which thus represent a therapeutic target in treatment of many
psychological diseases.
www.wjpps.com Vol 4, Issue 12, 2015.
933
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
CONCLUSION
In conclusion, this study revealed that oral administration of ethanolic extract of bark of
dolichondrone falcata and roots of scrophularia hypericifolia exhibit psychoactive property.
However in most of the activities dolichondrone falcata showed higher results compared to
scrophularia hypericifolia. The drugs showed greater results when used in combination. The
plant material should be further investigated for use in humans to treat brain disorder.
ACKNOWLEDGEMENT
I’m thankful to Mrs. Mehnoor Farheen M.Pharm, Ph.D. project guide who has continuously
helped and gave valuable suggestion to guide me in successful completion of the project.
With great pleasure, I acknowledge my profound and sincere thanks to Principal Mrs.
Sunitha, Shadan women’s college of pharmacy, Hyderabad for inspiring me, I wish to
express my gratitude to beloved vice principal Mrs. Nishath Farheen M.Pharm, Ph.D. Shadan
women’s college of pharmacy, Hyderabad for constant encouragement throughout the period,
which helped me in the successful completion of the project.
REFERENCES
1. Hascoét, M., Bourin, M. In Mood and Anxiety Related Phenotypes in Mice.
2. Dobson, K. S., & Joffe, R. (1986). The role of activity level and cognition in depressed
mood in a university sample. Journal of Clinical Psychology, 42(2): 264-271.
3. Pharmacographia India page 40.
4. Compendium of medicinal plants By Niir Board.
5. Hamza OJ, van den Bout-van den Beukel CJ, Matee MI, Moshi MJ, Mikx FH, Selemani
HO, et al. Antifungal activity of some Tanzanian plants used traditionally for the
treatment of fungal infections. J Ethnopharmacol., 2006; 108: 124–32.
6. Kokate CK. 4th ed. New Delhi: Vallabh Prakashan; 1994. Practical Pharmacognosy; p.
107.
7. Veeraraghavan, Prema Expert Consultant, CPCSEA, OECD Guideline No. 420. 2000
8. Chapter 11 in: Mitchell, Richard Sheppard; Kumar, Vinay; Abbas, Abul K.; Fausto,
Nelson. Robbins Basic Pathology. Philadelphia: Saunders. ISBN 1-4160-2973-7. 8th
edition.
9. Rodgers, R.J. (1997). "Animal models of anxiety: where next?” Behav. Pharmacol., 8:
477–496.
www.wjpps.com Vol 4, Issue 12, 2015.
934
Farheen et al. World Journal of Pharmacy and Pharmaceutical Sciences
10. Barraco RA, Coffin VL, Altman HJ, Phillis JW. Central effects on adenosine analogs on
locomotor activity in mice and antagonism of caffeine. Brain Res., 1983; 272: 392–395.
11. Dulawa SC, Hen R. Recent advances in animal models of chronic antidepressant effects:
The novelty-induced hypophagia test. Neurosci Biobehav Rev., 2005; 29(4–5): 771–83.
12. "Porsolt Forced Swim Test — Penn State University". Research.psu.edu. 2013-04-29.
Retrieved 2014-03-24.
13. Steru L, Chermat R, Thierry B, Simon P. The tail suspension test: a new method for
screening antidepressants in mice. Psychopharmacology (Berl)., 1985; 85: 367–370.
14. Dunham, M.W. and T.S. Miya, 1957. A note on a simple apparatus for detecting
neurological deficit in rats and mice. J. Am. Pharma. Assoc., 46: 208-209
15. Emamghoreishi M, Heidari-Hamedani G. Sedative-Hypnotic Activity of Extracts and
Essential Oil of Coriander Seeds. Iran. J. Med. Sci., 2006; 31: 22–27.
16. Koffer, K. B., Berney, S. & Hornykiewicz, O. Eur. J. Pharmac. 47, 81−86
(1978). | Article | ISI | ChemPort
17. Drug Discovery and Evaluation: Pharmacological Assays edited by Hans G. Vogel,
Wolfgang H. Vogel.
18. Takagi K, Watanabe M, Saito H. Studies on the spontaneous movement of animals by the
hole cross test: Effect of 2dimethylaminoethane. Its acylates on the central nervous
system. Jpn. J Pharmacol., 1971; 21: 797.
19. Boisser J R, Simon P. Dissociation dedeux compasanates dansle compartment
investigation de lasouris. Arch Int Pharmacodyn., 1964; 147: 372-88.
20. Madhu A, Keerthi PH, Jaideep S, Shivalinge GK. Antiepileptic activity of aqueous root
extract of Hemidesmus indicus in mice. Arch Pharm Sci Res., 2009; 1: 43–7.
21. Romanovsky, ―Thermoregulation: some concepts have changed. Functional architecture
of the thermoregulatory system,‖ American Journal of Physiology—Regulatory
Integrative and Comparative Physiology, vol. 292, no. 1, pp. R37–R46, 2007. View at
Publisher · View at Google Scholar · View at Scopus.
22. Allison DB, Casey DE. Antipsychotic-induced weight gain: a review of the literature. J
Clin Psychiatry., 2001; 62(Suppl 7): 22–31.
23. Randrup, A. and I. Munkvad, 1967. Stereotyped activities produced by amphetamine in
several animal species and man. Psychopharmacology., 11: 300-310.
24. Turner RA (1965): In: Screening Methods in Pharmacology., 113-117.