seminar.2016.Ph.D .4.pptx

NEUROPHARMACOLOGICAL EFFECTS
OF AQUEOUS EXTRACT AND
FRACTIONS OF ANCHOMANES
DIFFORMIS (BLUME) ENGLER
( ARACEAE) RHIZOME IN MICE
AGBOOLA, SAMUEL SUNDAY
B. Pharm.(Ife), M. Sc. (Ife)
(PHP11/12/H/2717)
NOVEMBER 2016
1

ABSTRACT
The search for effective and affordable drugs for the management of common central
nervous system disorders from drugs of natural origin has continued to attract the
attention of scientists all over the world. Many herbs have been claimed to be
efficacious in the treatment of these disorders by traditional herbal practitioners. Some
of these claims have been proved scientifically, thus justifying their ethnomedicinal
uses. The rhizome of Anchomanes difformis Blume Engl. (family Araceae) is claimed
to be used in the management of mental illness, among others, by herbalists in the
Southwestern Nigeria. However, no known detailed neuropharmacological study has
been done to verify this claim. This study carried out some neuropharmacological study
on the aqueous & hydro-alcoholic rhizome extracts and fractions in mice using animal
models for psychosis, anxiety, and depression. The results of acute toxicity test showed
LD50 values by oral route for the extracts and fractions as > 5000 mg/kg. The extract
significantly (p < 0.05) reduced the frequency of novelty induced rearing and
grooming; reduced period of immobility in Tail suspension and Forced swimming tests;
and increased both the frequency of entry and the time spent on the open arms of the
Elevated plus maze, thus showing promising anti-depressant and anxiolytic effects.
These results and other work done will be discussed in this seminar.
2

OUTLINE
• Introduction
• Objectives of the research
• Materials and methods
• Results/ Discussion
• Conclusion
• References
3

INTRODUCTION
Mental Illness
This is a disorder of the brain’s processes that makes
the patient feel ill, and may prevent him/her from
coping with daily life.
Examples include:
 Schizophrenia
 Anxiety
 Depression
4

INTRODUCTION: THE PLANT
ANCHOMANES DIFFORMIS (BLUME) ENGLER:
 Family name: Araceae
 Common names: Forest anchomanes (English)
‘Ogirisako’ or ‘Igo’ (Yoruba)
‘ Olumahi’ (Igbo)
‘Eba-enang’ (Efik)
5

Plate 1: A picture of A. difformis in its natural habitat
along Afao Road, Ado-Ekiti on 2nd May, 2013

7
Plate 2: A picture of a rhizome of A. difformis collected on
2nd May, 2013 along Afao Road, Ado-Ekiti, Nigeria

Ethnobotanical uses of A. difformis:
 Leaf & tuber:
Lactation stimulants (incl. veterinary)
 Sap:
Eye treatment; naso-pharyngeal infection
 Rhizome
Abortifacient, Treatment of asthma; gout, diuretic, laxative,
treatment of venereal diseases, diarrhoea, mental illness

INTRODUCTION: Pharmacological
studies reported on the plant
 Antimicrobial activity (Eneojo et al., 2011; Agyare et
al., 2016)
 Anti-trypanosomal activity (Atawodi et al., 2013)
 Analgesic and anti-inflammatory activities (Akah et
al., 1990; Agyare et al., 2016; Eke et al., 2013)
 Sedative activity (Eke et al., 2013)
 Anti-oxidant activity (Aliyu et al., 2013; Agyare et
al., 2016)
9

Table 1: Phytoconstituents of A.
difformis rhizome and leaf
Phytochemical
constituents
Rhizome Leaf
Tannins +++ +++
Alkaloids ++ +
Saponins +++ +++
Terpenoids ++ _
Cardiac glycosides +++ +++
Flavonoids ++ ++
10
Note: +++ = Conspicuously present; ++ = Moderately present; - = Absent
(Adapted with modification from Eneojo et al., 2011 )

Objectives of the study
a) determine the LD50 of the aqueous extract and fractions of the
rhizome of A. difformis
b) determine the effects of the extract and fractions of the rhizome
of A. difformis (Araceae) on novelty-induced behaviours in mice;
c) evaluate the anxiolytic, antidepressant, antipsychotic and
sedative/hypnotic effects of the extract and fractions; and
d) determine the mechanism(s) of action of the extract and the most
active fraction(s).
11

MATERIALS AND METHODS
 Plant materials
• Collection
• Identification and documentation
• Extraction & fractionation
• Preparation of sample solution & reference drugs
 Experimental animals
• Swiss albino mice of either sex (18-25 g)
 Drugs used: Apomorphine, Cyproheptadine, Diazepam, Flumazenil,
Haloperidol, Imipramine, Pentobarbital, Sertraline, Yohimbine, Normal
saline, Tween 80.
12

METHODS
 CNS effects assessment of Aqueous rhizome extract and fractions:
• LD50 determination by Lorke’s Method (Lorke, 1983)
• Sedative: OFT(Crawley, 1985) & Pentobarbital-induced hypnotic
models (Erden et al., 2001, Olayiwola et al., 2013)
• Anxiolytic: EPM (Trullas & Skolnick, 1993; Akanmu et al., 2011),
Staircase (Simiand et al., 1984) & Hole-board models
(File & Pellow, 1985; Nic Dhonnchadha et al., 2003)
• Antidepressant: FST (Porsolt et al., 1977) & TST models (Steru et
al., 1985)
• Antipsychotic: Swim-induced grooming (Kedves et al., 2008),
Apomophine-induced stereotypy model (Siqueira et
al., 1998). 13

STATISTICALANALYSIS
 The results are presented as the mean (n=5) ±
standard error of mean(SEM).
 The values were compared using the One-way
analysis of variance (ANOVA) followed by
Student-Newman-Keuls post hoc test.
 *significant p< 0.05 versus vehicle-treated control
 #significant p<0.05 versus standard drug 14

RESULTS/DISCUSSION
ADE = Aqueous extract of A. difformis rhizome; HF = n-Hexane fraction; EF = Ethylacetate fraction;
BF = Butanol fraction; AF = Residual aqueous fraction of hydro-alcoholic extract of A. difformis
rhizome.
*0/3 means the mice did not die 15
DOSE
LEVEL
MORTALITY WITHIN 24 HOURS
(n=3)
% MORTALITY WITHIN 24
HOURS
ADE HF EF BF AF ADE HF EF BF AF
10 mg/kg 0/3 0/3 0/3 0/3 0/3 0.00 0.00 0.00 0.00 0.00
100 mg/kg 0/3 0/3 0/3 0/3 0/3 0.00 0.00 0.00 0.00 0.00
1000 mg/kg 0/3 0/3 0/3 0/3 0/3 0.00 0.00 0.00 0.00 0.00
Table 2: Acute toxicity (mortality) test (Lorke’s Method) for extracts and fractions of A.
difformis in mice (oral route): Phase I

Table 3: Acute Toxicity Study-phase II (Oral)
(Lorke’s Method)
ADE = Aqueous extract of A. difformis rhizome; HF = n-Hexane fraction; EF = Ethylacatate fraction; BF
= Butanol fraction; AF = Residual aqueous fraction of hydro-alcoholic extract of A. difformis rhizome.
*0/1 means the mouse did not die
16
DOSE
LEVEL
MORTALITY WITHIN 24
HOURS (n=3)
% MORTALITY WITHIN 24
HOURS
ADE HF EF AF ADE HF EF BF AF
1000 mg/kg 0/1* 0/1 0/1 0/1 0.00 0.00 0.00 0.00 0.00
1600 mg/kg 0/1 0/1 0/1 0/1 0.00 0.00 0.00 0.00 0.00
2900 mg/kg 0/1 0/1 0/1 0/1 0.00 0.00 0.00 0.00 0.00
5000 mg/kg 0/1 0/1 0/1 0/1 0.00 0.00 0.00 0.00 0.00

Fig 1: Effect of ADE on novelty-induced grooming behaviour in mice
ADE: Aqueous extract of A. difformis rhizome; DZP: Diazepam (4 mg/kg,
i.p.); VEH: Vehicle (Normal saline, 10 ml/kg, p.o.); *significant p < 0.05
compared to vehicle; #significant p < 0.05 compared to the standard drug,
Diazepam.
17
0
5
10
15
20
25
30
35
40
VEH 30 60 125 250 500 1000 DZP
Frequency
of
grooming/20
min
ADE (mg/kg, p.o)
*
*
*
*
*
#
#
#
# #
*
*
#
NIB: GROOMING

18
0
5
10
15
20
25
30
35
40
VEH 30 60 125 250 500 1000 DZP
Frequency
of
grooming/20
min
HF (mg/kg, p.o.)
*
*
*
*
*
*
#
#
#
#
#
#
Fig. 3: Effect of HF of A. difformis rhizome on novelty-induced grooming
behaviour in mice
HF: Hexane fraction of A. difformis rhizome of hydro-alcoholic extract; DZP:
Diazepam, 4 mg/kg, i.p.; VEH: Vehicle (i.e. 10 ml/kg 2.5% Tween 80 in saline, p.o.);
*significant p < 0.05 compared to vehicle; #significant p < 0.05 compared to reference
drug, Diazepam.
NIB: GROOMING

Fig. 4: Effect of EF of Anchomanes difformis on novelty-induced grooming in
mice. EF = Doses of Ethanol fraction of A. difformis rhizome; DZP = Diazepam (4
mg/kg, i.p.); VEH = Vehicle (i.e. 2.5 % Tween 80 in saline, 10 ml/ kg, p.o.);
*significant p < 0.05 compared to vehicle; #significant p < 0.05 compared to
reference drug, Diazepam. 19
0
5
10
15
20
25
30
35
40
VEH 30 60 125 250 500 1000 DZP
Frequency
of
grooming/20
min
EF (mg/kg, p.o.)
*
* *
*
# #
#
#
# #
NIB: GROOMING

NIB: GROOMING
20
0
5
10
15
20
25
30
35
40
VEH 30 60 125 250 500 1000 DZP
Frequency
of
grooming/20
min
BF (mg/kg, p.o.)
*
#
*
#
*
#
#
#
#
*
Fig. 5: Effect of BF of Anchomanes difformis on novelty-induced grooming in mice.
BF = Doses of Buthanol fraction of A. difformis rhizome; DZP = Diazepam (4 mg/kg, i.p.);
VEH = Vehicle (i.e. 2.5 % Tween 80 in saline, 10 ml/ kg, p.o.); *significant p < 0.05
compared to vehicle; #significant p < 0.05 compared to reference drug, Diazepam.

Fig. 6: Effect of Anchomanes difformis on novelty-induced grooming in mice.
AF = Doses of residual aqueous fraction of A. difformis rhizome; DZP =
Diazepam (4 mg/kg, i.p.); VEH = Vehicle (i.e. Normal saline, 10 ml/ kg, p.o.);
*significant p < 0.05 compared to vehicle; #significant p < 0.05 compared to
reference drug, Diazepam. 21
0
5
10
15
20
25
30
35
40
VEH 30 60 125 250 500 1000 DZP
Frequency
of
grooming/20
min
AF (mg/kg, p.o.)
*
*
*
#
#
#
#
#
#
*
NIB: GROOMING

NIB: REARING
22
0
10
20
30
40
50
60
70
80
90
VEH 30 60 125 250 500 1000 DZP
Frequency
of
rearing/20
min
ADE (mg/kg, p.o.)
*
*
*
#
#
#
#
#
#
Fig. 7: Effect of ADE on novelty-induced rearing behaviour in mice
ADE: Aqueous extract of A. difformis rhizome; DZP: Diazepam (4 mg/kg, i.p.);
VEH: Vehicle (Normal saline, 10 ml/kg, p.o.); *significant p < 0.05 compared to
vehicle; #significant p < 0.05 compared to the standard drug, Diazepam.

NIB: REARING
23
0
20
40
60
80
100
120
140
VEH 30 60 125 250 500 1000 DZP
Frequency
of
rearing/20
min
HF (mg/kg, p.o.)
* *
*
* *
*
*
#
#
#
# #
#
Fig. 9: Effect of HF on novelty-induced rearing behaviour in mice
HF: Hexane fraction of A. difformis rhizome; DZP: Diazepam (4 mg/kg, i.p.); VEH: Vehicle
(2.5 % Tween 80 in saline, 10 ml/kg, p.o.); *significant p < 0.05 compared to vehicle;
#significant p < 0.05 compared to the standard drug, Diazepam.

NIB: REARING
24
Fig. 10: Effect of EF on novelty-induced rearing behaviour in mice
EF: Ethylacetate fraction of A. difformis rhizome; DZP: Diazepam (4 mg/kg, i.p.);
VEH: Vehicle (2.5 % Tween 80 in saline, 10 ml/kg, p.o.); *significant p < 0.05
Diazepam.
0
20
40
60
80
100
120
140
VEH 30 60 125 250 500 1000 DZP
Frequency
of
rearing/20
min
EF (mg/kg, p.o.)
*
*
*
*
#
#
#
#
#
#

NIB: REARING
25
0
20
40
60
80
100
120
140
VEH 30 60 125 250 500 1000 DZP
Frequency
of
rearing/20
min
BF (mg/kg, p.o.)
*
*
# #
#
#
#
#
Fig. 11: Effect of BF on novelty-induced rearing behaviour in mice
BF: Butanol fraction of A. difformis rhizome; DZP: Diazepam (4 mg/kg, i.p.); VEH: Vehicle
(2.5 % Tween 80 in saline, 10 ml/kg, p.o.); *significant p < 0.05 compared to vehicle;

NIB: REARING
26
0
10
20
30
40
50
60
70
80
90
VEH 30 60 125 250 500 1000 DZP
Frequency
of
rearing/20
min
AF (mg/kg, p.o.)
*
*
#
#
#
#
#
#
Fig. 12:Effect of AF on novelty-induced rearing behaviour in mice
AF: Residual aqueous fraction of A. difformis rhizome; DZP: Diazepam (4 mg/kg, i.p.); VEH:
Vehicle (Normal saline, 10 ml/kg, p.o.); *significant p < 0.05 compared to vehicle;

NIB: LINE-CROSSING
27
Fig. 13: Effect of ADE on novelty-induced horizontal locomotion behaviour
in mice
ADE: Aqueous extract of A. difformis rhizome; DZP: Diazepam (4 mg/kg,
i.p.); VEH: Vehicle (Normal saline, 10 ml/kg, p.o.); *significant p < 0.05
Diazepam.
0
50
100
150
200
250
300
350
VEH 30 60 125 250 500 1000 DZP
Frequency
of
line
crossing/20
min
ADE (mg/kg, p.o.)
*
*
*
#
# # #

NIB: LINE-CROSSING
28
0
50
100
150
200
250
300
350
400
VEH 30 60 125 250 500 1000 DZP
Frequency
of
line
crossing/20
min
HF (mg/kg, p.o.)
* *
*
*
*
*
*
# #
Fig. 15: Effect of HF on novelty-induced horizontal locomotion (line-crossing)
in mice
HF: N-hexane fraction of A. difformis rhizome; DZP: Diazepam (4 mg/kg, i.p.);
VEH: Vehicle (2.5% Tween 80 in saline, 10 ml/kg, p.o.); *significant p < 0.05
compared to vehicle (control); #significant p < 0.05 compared to the standard drug,
Diazepam.

NIB: LINE-CROSSING
29
0
50
100
150
200
250
300
350
400
VEH 30 60 125 250 500 1000 DZP
Frequency
of
line
crossing/20
min
EF (mg/kg, p.o.)
*
* *
*
#
#
#
#
#
*
Fig. 16: Effect of EF on novelty-induced horizontal locomotion (line-crossing) in
mice
EF: Ethyl-acetate fraction of A. difformis rhizome; DZP: Diazepam (4 mg/kg, i.p.); VEH:
Vehicle (2.5% Tween 80 in saline, 10 ml/kg, p.o.); *significant p < 0.05 compared to
vehicle (control); #significant p < 0.05 compared to the standard drug, Diazepam.

NIB: LINE CROSSING
30
0
50
100
150
200
250
300
350
400
VEH 30 60 125 250 500 1000 DZP
Frequency
of
line
crossing/20
min
BF (mg/kg, p.o.)
*
*
#
#
#
# # #
Fig. 17: Effect of BF on novelty-induced horizontal locomotion (Line-crossing) in mice
BF: Butanol fraction of A. difformis rhizome; DZP: Diazepam (4 mg/kg, i.p.); VEH: Vehicle
(2.5% Tween 80 in saline, 10 ml/kg, p.o.); *significant p < 0.05 compared to vehicle
(control); #significant p < 0.05 compared to the standard drug, Diazepam.

HYPNOTIC TEST
31
0
20
40
60
80
100
120
140
160
180
VEH 250 500 1000 DZP
Sleeping
time
(min)
ADE (mg/kg, p.o.)
*
*
#
#
#
Fig. 19: Effects of ADE on pentobarbital-induced sleep latency (Panel A) and sleeping
time (Panel B) in mice
ADE: Aqueous extract of A. difformis rhizome; DZP: Diazepam (2 mg/kg, i.p.); VEH:
Vehicle (Normal saline, 10 ml/kg, p.o.).* p < 0.05 significantly different from vehicle
(control); # p < 0.05 significantly different from standard drug, Diazepam.
0
1
2
3
4
5
6
VEH 250 500 1000 DZP
Sleep
latency
(min)
ADE (mg/kg, p.o.)
*
*
*
#
#
#

HYPNOTIC TEST
32
0
1
2
3
4
5
6
VEH 250 500 1000 DZP
Sleep
latency
(min)
HF (mg/kg, p.o.)
*
*
*
#
#
*
#
0
20
40
60
80
100
120
140
160
VEH 250 500 1000 DZP
Sleeping
time(min)
HF (mg/kg, p.o.)
*
*
*
*
#
+
Pent
o
Panel A
Panel B
Fig. 21: Effects of HF on pentobarbital-induced sleep latency (Panel A) and sleeping
HF: N-hexane fraction of A. difformis rhizome; DZP: Diazepam (2 mg/kg, i.p.); VEH:
Vehicle (2.5% Tween 80 in saline, 10 ml/kg, p.o.).* p < 0.05 significantly different from
vehicle (control); # p < 0.05 significantly different from standard drug, Diazepam.

HYPNOTIC TEST
• Fig. 22: Effect of EF on
Pentobarbital-induced
sleep latency (Panel A) and
sleeping time (Panel B) in
mice
• EF: Ethyl-acetate fraction of
A. difformis rhizome;
• DZP: Diazepam (2 mg/kg,
i.p.);
• VEH: Vehicle (2.5% Tween
80 in saline, 10 ml/kg, p.o.);
• *significant p < 0.05
compared to vehicle
(control);
• #significant p < 0.05
compared to standard drug,
Diazepam.
33
0
1
2
3
4
5
6
Sleep
latency
(min
)
*
*
# # #
0
20
40
60
80
100
120
140
160
VEH 250 500 1000 DZP
Sleeping
time
(min)
EF (mg/kg)
*
* *
*
#
Panel A
Panel B

HYPNOTIC TEST
• Fig. 23: Effects of BF on
Pentobarbital-induced sleep
latency (Panel A) and
mice
• BF: Butanol fraction of A.
difformis rhizome;
i.p.);
• *significant p < 0.05 compared
to vehicle (control);
• #significant p < 0.05 compared
to standard drug, Diazepam.
34
0
1
2
3
4
5
6
Sleep
latency
(min)
*
#
#
#
0
50
100
150
200
VEH 250 500 1000 DZP
Sleeping
time
(min)
BF (mg/kg)
*
*
#
#
#
Panel A
Panel B

HYPNOTIC TEST
• Fig. 24: Effects of AF on
Pentobarbital-induced
sleep latency (Panel A) and
mice
• AF: Aqueous fraction of A.
difformis rhizome;
i.p.);
• VEH: Vehicle (Normal
saline, 10 ml/kg, p.o.);
• *significant p < 0.05
compared to vehicle
(control);
• #significant p < 0.05
compared to standard drug,
Diazepam.
35
Panel A
0
1
2
3
4
5
Sleep
latency
(min
)
*
#
#
#
Panel B
0
20
40
60
80
100
120
140
160
180
VEH 250 500 1000 DZP
Sleeping
time
(min)
AF (mg/kg)
*
*
#
#
#

Mechanism: Hypnotic effect
• Fig. 25: Influence of Flumazenil (3
mg/kg, i.p.) on effect of ADE (1000
mg/kg, p.o.) on Pentobarbital-
induced sleep latency (Panel A) and
sleeping time (Panel B) in mice
• ADE: Aqueous extract of A. difformis
rhizome (1000 mg/kg, p.o.);
• VEH: Vehicle-treated control group
(Normal saline, 10 ml/kg, p.o.);
• DZP: Diazepam (2 mg/kg, i.p.);
Flum: Flumazenil (3 mg/kg, i.p.);
• #significant p < 0.05 : antagonist
versus treatment alone.
36
Panel A
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
Sleep
latency
(min)
#
#
Panel B
0
20
40
60
80
100
120
140
160
180
VEH Flum VEH Flum VEH Flum
Sleeping
time
(min)
ADE
#
#
DZP

Mechanism: Hypnotic effect
• Fig. 27: Influence of Flumazenil
(3 mg/kg, i.p.) on effect of HF
(1000 mg/kg, p.o.) on
Pentobarbital-induced sleep
latency (Panel A) and sleeping
• HF: N-Hexane fraction of A.
difformis rhizome (1000 mg/kg,
p.o.);
• DZP: Diazepam (2 mg/kg, i.p.);
• VEH: Vehicle (2.5% Tween 80 in
• Flum: Flumazenil (3 mg/kg. i.p.);
#significant p < 0.05: antagonist
37
Panel A
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
Sleep
latency
(min)
ns
#
Panel B
0
50
100
150
200
Sleeping
time
(min)
HF
# #
DZP

Anxiolytic Test: EPM
• Fig. 28: Effect of ADE on %
open arm entries (Panel A)
and % time spent in the open
arm (Panel B) in mice
• ADE: Aqueous extract of A.
difformis rhizome;
• DZP: Diazepam (0.5 mg/kg,
i.p.);
• VEH: Vehicle (Normal saline,
10 ml/kg, p.o.); *significant p
< 0.05 compared to vehicle
(control);
to standard drug, Diazepam.
38
Panel A
0
10
20
30
40
50
60
70
%
Open
arm
entries/5
min
*
* *
*
*
Panel B
0
10
20
30
40
50
60
70
VEH 30 60 125 250 DZP
%
Open
arm
duration
/5
min
ADE (mg/kg, p.o.)
*
#
#
*

• Fig. 30: Effects of HF on %
open arm entries (Panel A) and
% time spent in the open arm
(Panel B) in mice
• HF: N-hexane fraction of A.
difformis rhizome;
• DZP: Diazepam (0.5 mg/kg, i.p.);
• *significant p < 0.05 compared to
vehicle (control);
• #significant p < 0.05 compared to
standard drug, Diazepam.
39
Panel A
0
10
20
30
40
50
60
70
%
Open
arm
entries
/5
min
*
* *
*
*
Panel B
0
20
40
60
80
VEH 30 60 125 250 DZP
%
Open
arm
duration/5
min
HF (mg/kg, p.o.)
*
*

• Fig. 31: Effect of EF on %
• EF: Ethyl-acetate fraction of A.
difformis rhizome;
i.p.);
to standard drug, Diazepam
40
Panel A
0
10
20
30
40
50
60
70
%
Open
arm
entries/5
min
* * *
Panel B
0
20
40
60
80
100
VEH 30 60 125 250 DZP
%
Open
arm
duration/5
min
EF (mg/kg, p.o.)
*
* *

• Fig. 32: Effects of BF on %
difformis rhizome;
i.p.);
• VEH: Vehicle (2.5% Tween 80
in saline, 10 ml/kg, p.o.);
41
Panel A
0
20
40
60
80
%
Open
arm
entries/5
min
*
#
# #
Panel B
0
20
40
60
80
VEH 30 60 125 250 DZP
%
Open
arm
duration/5
min
BF (mg/kg, p.o.)
*
*
*
#
#

• Fig. 33: Effects of AF on %
and % time spent in the
open arm (Panel B) in mice
difformis rhizome;
i.p.);
• VEH: Vehicle (Normal saline,
10 ml/kg, p.o.);
42
Panel A
0
20
40
60
80
%
Open
arm
entries/5
min
*
* *
*
#
#
#
Panel B
0
10
20
30
40
50
60
70
VEH 30 60 125 250 DZP
%
Open
arm
duration
/5
min
AF (mg/kg, p.o.)
*
*
*
#
#

Open Arm Avoidance Index
43
0
10
20
30
40
50
60
70
80
VEH 30 60 125 250 DZP
Open
Arm
Avoidance
Index
ADE (mg/kg, p.o.)
Fig. 34: Effect of ADE on OAAI in mice
ADE: Aqueous extract of A. difformis rhizome; DZP: Diazepam (0.5 mg/kg,
i.p.); VEH: Vehicle (Normal saline, 10 ml/kg, p.o.); OAAI: Open Arm
Avoidance Index; Broken lines (…….) indicate the cut-off point which is 10
points below the OAAI for the vehicle-treated control.

44
0
10
20
30
40
50
60
70
80
VEH 30 60 125 250 DZP
Open
Arm
Avoidance
Index
HF (mg/kg, p.o.)
Fig. 36: Effect of HF on OAAI in mice
HF: N-hexane fraction of A. difformis rhizome; DZP: Diazepam (0.5 mg/kg, i.p.);
VEH: Vehicle (2.5% Tween 80 in saline, 10 ml/kg, p.o.); OAAI: Open Arm

45
0
10
20
30
40
50
60
70
80
VEH 30 60 125 250 DZP
Open
Arm
Avoidance
Index
EF (mg/kg, p.o.)
Fig. 37: Effect of EF on OAAI in mice
EF: Ethyl-acetate fraction of A. difformis rhizome; DZP: Diazepam (0.5 mg/kg, i.p.); VEH:
Vehicle (2.5% Tween 80 in saline, 10 ml/kg, p.o.); OAAI: Open Arm Avoidance Index;
Broken lines (…….) indicate the cut-off point which is 10 points below the OAAI for the
vehicle-treated control.

46
0
10
20
30
40
50
60
70
80
VEH 30 60 125 250 DZP
Open
Arm
Avoidance
Index
BF (mg/kg, p.o.)
Fig. 38: Effect of BF on OAAI in mice
BF: Butanol fraction of A. difformis rhizome; DZP: Diazepam (0.5 mg/kg, i.p.);
VEH: Vehicle (2.5% Tween 80 in saline, 10 ml/kg, p.o.); OAAI: Open Arm

47
0
10
20
30
40
50
60
70
80
VEH 30 60 125 250 DZP
Open
Arm
Avoidance
Index
AF (mg/kg, p.o.)
Fig. 39: Effect of AF on OAAI in mice
AF: Aqueous fraction of A. difformis rhizome; DZP: Diazepam (0.05 mg/kg, i.p.);
VEH: Vehicle (Normal saline, 10 ml/kg, p.o.); OAAI: Open Arm Avoidance
Index; Broken lines (…….) indicate the cut-off point which is 10 points below the
OAAI for the vehicle-treated control.

Anxiolytic test: Hole-board
48
0
5
10
15
20
25
30
VEH 30 60 125 250 DZP
No
of
head-dips/5
min
ADE (mg/kg, p.o.)
*
*
# #
#
Fig. 40: Effect of ADE on the number of head-dips made by mice on the Hole-board
ADE: Aqueous extract of A. difformis rhizome;
DZP: Diazepam (0.5 mg/kg, i.p.);
VEH: Vehicle (Normal saline, 10 ml/kg, p.o.);
*significant p < 0.05 compared to vehicle (control);
#significant p < 0.05 compared to standard drug, Diazepam.

49
0
5
10
15
20
25
30
35
40
45
50
VEH 30 60 125 250 DZP
No
of
head-dips/5
min
HF (mg/kg, p.o.)
*
*
*
*
*
Fig. 42: Effect of HF on the number of head-dips made by mice on the Hole-
board
HF: N-hexane fraction of A. difformis rhizome;
VEH: Vehicle (2.5% Tween 80 in saline, 10 ml/kg, p.o.);
*significant p < 0.05 compared to vehicle (control).

50
0
5
10
15
20
25
30
35
40
VEH 30 60 125 250 DZP
No
of
head-dips/5
min
EF (mg/kg, p.o.)
*
* *
*
*
Fig. 43: Effect of EF on the number of head-dips made by mice on the Hole-
board
EF: Ethyl-acetate fraction of A. difformis rhizome;
*significant p < 0.05 compared to vehicle-treated control.

51
Fig. 44: Effect of BF on the number of head-dips made by mice on the
Hole-board
BF: Butanol fraction of A. difformis rhizome;
*significant p < 0.05 compared to vehicle-treated control;
#significant p < 0.05 compared to Diazepam, 0.5 mg/kg, i.p.
0
5
10
15
20
25
30
35
VEH 30 60 125 250 DZP
No
of
head-dips/5
min
BF (mg/kg, p.o.)
*
# #

52
Fig. 45: Effect of AF on the number of head-dips made by mice on the hole-
board
AF: Aqueous fraction of A. difformis rhizome;
DZP: Diazepam (0.05 mg/kg, i.p.); VEH:
Vehicle (Normal saline, 10 ml/kg, p.o.);
*significant p < 0.05 compared to vehicle (control);
#significant p < 0.05 compared to standard drug, Diazepam.
0
5
10
15
20
25
30
VEH 30 60 125 250 DZP
No
of
head-dips/5
min
AF (mg/kg, p.o.)
*
#
# # #

Anxiolytic test: Staircase Model (SCM)
53
0
10
20
30
40
50
No
of
steps
climbed
/5
min
*
*
#
#
#
#
Panel A
Panel B
0
5
10
15
20
VEH 30 60 125 250 DZP
Frequency
of
rearing
/5
min
ADE (mg/kg, p.o.)
*
*
*
*
*
Fig. 46: Effects of ADE on the number of steps climbed (Panel A) and the frequency
of rearing (Panel B) by mice
ADE: Aqueous extract of A. difformis rhizome; DZP: Diazepam (0.5 mg/kg, i.p.); VEH:
Vehicle (Normal saline, 10 ml/kg, p.o.); * significant p < 0.05 compared to vehicle
(control); #significant p < 0.05 compared to standard drug, Diazepam.

Anxiolytic test: Staircase Model
54
Panel A
0
10
20
30
40
50
No
of
steps
climbed
/5
min
*
#
#
#
#
Panel B
0
5
10
15
20
25
30
VEH 30 60 125 250 DZP
Frequency
of
rearing/5
min
HF (mg/kg, p.o.)
* *
*
*
Fig. 48: Effects of HF on the number of steps climbed (Panel A) and the frequency of
rearing (Panel B) by mice
HF: N-hexane fraction of A. difformis rhizome; DZP: Diazepam (0.5 mg/kg, i.p.); VEH:
Vehicle (2.5% Tween 80 in saline, 10 ml/kg, p.o.);* significant p < 0.05 compared to vehicle-
treated control;# significant p < 0.05 compared to standard drug, Diazepam.

55
Panel A
0
5
10
15
20
25
30
35
40
45
No
of
steps
climbed
/5
min
*
#
#
#
#
Panel B
0
5
10
15
20
25
30
VEH 30 60 125 250 DZP
Frequency
of
rearing
/5
min
EF (mg/kg, p.o.)
* *
*
*
*
Fig. 49: Effects of EF on the number of steps climbed (Panel A) and the frequency of
EF: Ethyl-acetate fraction of A. difformis rhizome; DZP: Diazepam (0.5 mg/kg, i.p.);
VEH: Vehicle (2.5% Tween 80 in saline, 10 ml/kg, p.o.); * significant p < 0.05 compared to
vehicle-treated control; #significant p < 0.05 compared to standard drug, Diazepam.

56
Panel A
0
10
20
30
40
50
No
of
steps
climbed
/5
min
*
#
#
#
#
Panel B
0
5
10
15
20
25
30
VEH 30 60 125 250 DZP
Frequency
of
rearing/5
min
BF (mg/kg, p.o.)
*
*
* *
*
Fig. 50: Effects of BF on the number of steps climbed (Panel A) and the frequency of
BF: Butanol fraction of A. difformis rhizome; DZP: Diazepam (0.5 mg/kg, i.p.); VEH: Vehicle
(2.5% Tween 80 in saline, 10 ml/kg, p.o.); *significant p < 0.05 compared to vehicle-treated
control; #significant p < 0.05 compared to standard drug, Diazepam.

57
Panel A
0
10
20
30
40
50
No
of
steps
climbed/5
min
*
#
#
#
#
Panel B
0
5
10
15
20
VEH 30 60 125 250 DZP
Frequency
of
rearing/5
min
AF (mg/kg, p.o.)
*
*
Fig. 51: Effects of AF on the number of steps climbed (Panel A) and the frequency of
BF: Aqueous fraction of A. difformis rhizome; DZP: Diazepam (0.5 mg/kg, i.p.); VEH:
Vehicle (Normal saline, 10 ml/kg, p.o.); *significant p < 0.05 compared to vehicle-treated
control; #significant p < 0.05 compared to standard drug, Diazepam.

Mechanism of Anxiolytic effect: GABAA receptor involvement (EPM)
• Fig. 52: : Influence of Flumazenil (3
mg/kg, i.p.) on effects of ADE (30
mg/kg, p.o.) on % entries into open
arm (Panel A) and % time spent on
open arm (Panel B) of EPM in mice
• VEH: Vehicle (Normal saline, 10
ml/kg, p.o.);
• Flum: Flumazenil (3 mg/kg, i.p.);
• #significant p < 0.05: antagonist
58
Panel A
0
10
20
30
40
50
60
70
%
Open
Arm
entry
#
ns
Panel B
0
10
20
30
40
50
60
70
%
Open
Arm
duration
#
#
ADE DZP

Mechanism of Anxiolytic effect: GABAA receptor involvement (EPM)
• Fig. 54: Influence of Flumazenil
(3 mg/kg, i.p.) on effects of EF
(125 mg/kg, p.o.) on % entries
into open arm (Panel A) and %
time spent on open arm (Panel
B) of EPM in mice
difformis rhizome (125 mg/kg,
p.o.);
• Flum: Flumazenil (3 mg/kg, i.p.);
59
Panel A
0
10
20
30
40
50
60
70
%
Open
Arm
entry
# #
Panel B
0
10
20
30
40
50
60
70
%
Open
Arm
duration
#
#
EF DZP

Mechanism of Anxiolytic effect: Serotonergic/histaminergic receptor
involvement (EPM)
• Fig. 55: Influence of Cyproheptadine
(0.5 mg/kg, i.p.) on effect of ADE (30
mg/kg, p.o.) on % entries into open
arm (Panel A) and % time spent on
open arm (Panel B) of EPM in mice
• VEH: Vehicle (Normal saline, 10 ml/kg,
p.o.);
• CYP: Cyproheptadine (0.5 mg/kg, i.p.);
• STR: Sertraline (20 mg/kg, p.o.);
• #significant p < 0.05: antagonist versus
treatment alone.
60
Panel A
0
10
20
30
40
50
60
%
Open
Arm
entry
# #
Panel B
0
10
20
30
40
50
60
70
80
VEH CYP VEH CYP VEH CYP
%
Open
Arm
duration
# #
ADE STR

involvement (EPM)
• Fig. 57: Influence of
Cyproheptadine (0.5 mg/kg, i.p.)
on effects of EF (125
mg/kg)/STR (20 mg/kg) on %
open arm entries (Panel A) and
% time spent (Panel B) by mice
on EPM
difformis rhizome;
• CYP: Cyproheptadine (0.5 mg/kg,
i.p.);
61
Panel A
0
10
20
30
40
50
60
%
Open
Arm
entry
#
#
Panel B
0
10
20
30
40
50
60
70
80
%
Open
Arm
duration
# #
STR
EF

Mechanism of Anxiolytic effect: GABAA receptor involvement(Hole-board)
62
0
5
10
15
20
25
30
Number
of
head-dips/5
min #
ADE DZP
#
Fig. 58: Influence of Flumazenil (3 mg/kg, i.p.) on the number of head-dips induced
by ADE (30 mg/kg, p.o.) in mice
ADE: Aqueous extract of A. difformis rhizome (30 mg/kg, p.o.);
Flum: Flumazenil (3 mg/kg, i.p.);
#significant p < 0.05: antagonist versus treatment alone.

Mechanism of Anxiolytic effect: GABAA receptor involvement(Hole-board)
63
0
5
10
15
20
25
30
35
40
Number
of
head-dips/5
mins
#
#
EF DZP
Fig. 60: Influence of Flumazenil (3 mg/kg, i.p.) on the number of head-dips induced by EF
(125 mg/kg, p.o.) in mice on the Hole-board
EF: Ethyl-acetate fraction of A. difformis rhizome;
Flum: Flumazenil (3 mg/kg. i.p.); #significant p < 0.05: antagonist versus treatment alone.

Mechanism of Anxiolytic effect: Serotonergic/histaminergic receptor involvement
(Hole-board)
64
0
10
20
30
40
50
60
VEH CYP VEH CYP STR CYP
Number
of
head-dips/5
min
#
#
ADE STR
Fig. 61: Influence of Cyproheptadine (0.5 mg/kg, i.p.) on the number of head-dips
induced by ADE (30 mg/kg, p.o.) in mice on the Holeboard
ADE: Aqueous extract of A. difformis rhizome (30 mg/kg, p.o.);
CYP: Cyproheptadine (0.5 mg/kg, i.p.);
STR: Sertraline (20 mg/kg, p.o.);

involvement
(Hole-board)
65
0
10
20
30
40
50
60
Number
of
head-dips/5
min
#
#
STR
EF
Fig. 63: Influence of Cyproheptadine (0.5 mg/kg, i.p.) on the number of head-dips
induced by EF (125 mg/kg, p.o.) in mice on the Hole-board
EF: Ethyl-acetate fraction of A. difformis rhizome (125 mg/kg, p.o.);
STR: Sertraline (20 mg/kg, p.o.);
CYP: Cyproheptadine (0.5 mg/kg, i.p.);

Antidepressant study: TST & FST
66
Panel A
0
100
200
300
Period
of
immobility
(s)
* *
*
#
#
#
#
*
Panel B
0
50
100
150
200
VEH 60 125 250 500 1000 IMP
Period
of
immobility
(s)
ADE (mg/kg, p.o.)
*
*
#
#
#
#
#
*
Fig. 64: Effects of ADE on period of immobility in mice on TST (Panel A) and FST
(Panel B)
ADE: Aqueous extract of A. difformis rhizome; IMP: Imipramine (25 mg/kg, i.p.); VEH:
Vehicle (Normal saline, 10 ml/kg, p.o.); *significant p < 0.05 compared to vehicle (control);
#significant p < 0.05 compared to standard drug, Imipramine.

67
Panel A
0
100
200
300
Period
of
immobility
(s)
*
#
#
#
#
#
Panel B
0
50
100
150
200
VEH 60 125 250 500 1000 IMP
Period
of
immobility
(s)
HF (mg/kg, p.o.)
*
# # # # #
Fig. 66: Effects of HF on period of immobility in mice on TST (Panel A) and FST (Panel
B)
HF: N-hexane fraction of A. difformis rhizome; IMP: Imipramine (25 mg/kg, i.p.); VEH:
Vehicle (2.5% Tween 80 in saline, 10 ml/kg, p.o.);
*significant p < 0.05 compared to vehicle-treated control; #significant p < 0.05 compared to
standard drug, Imipramine.

68
Panel A
0
50
100
150
200
Period
of
immobility
(s)
* *
*
#
*
#
#
#
#
Panel B
0
50
100
150
200
VEH 60 125 250 500 1000 IMP
Period
of
immobility
(s)
EF (mg/kg, p.o.)
*
*
*
*
#
#
# #
#
Fig. 67: Effects of EF on period of immobility in mice on TST (Panel A) and FST (Panel
B)
EF: Ethyl-acetate fraction of A. difformis rhizome; IMP: Imipramine (25 mg/kg, i.p.); VEH:
Vehicle (2.5% Tween 80 in saline, 10 ml/kg, p.o.); *significant p < 0.05 compared to vehicle-
treated control; #significant p < 0.05 compared to standard drug, Imipramine

69
Panel A
0
50
100
150
200
Period
of
immobility
(s
)
*
# #
# # #
Panel B
0
50
100
150
200
VEH 60 125 250 500 1000 IMP
Period
of
immobility
(s
)
BF (mg/kg, p.o.)
*
# #
#
#
#
*
*
Fig. 68: Effects of BF on period of immobility in mice on TST (Panel A) and FST (Panel
B)
BF: Butanol fraction of A. difformis rhizome; IMP: Imipramine (25 mg/kg, i.p.); VEH: Vehicle
(2.5% Tween 80 in saline, 10 ml/kg, p.o.). *significant p < 0.05 compared to vehicle (control);
#significant p < 0.05 compared to standard drug, Imipramine

70
Panel A
0
50
100
150
200
250
Period
of
immobility
(s)
*
#
#
#
#
#
Panel B
0
50
100
150
200
250
VEH 60 125 250 500 1000 IMP
Period
of
immobility
(s)
AF (mg/kg, p.o.)
*
*
#
*
#
*
#
*
#
*
#
Fig. 69: Effects of AF on period of immobility in mice on TST (Panel A) and FST
(Panel B)
AF: Residual Aqueous fraction of A. difformis rhizome; IMP: Imipramine (25 mg/kg, i.p.);
VEH: Vehicle (Normal saline, 10 ml/kg, p.o.). *significant p < 0.05 compared to vehicle-
treated control; #significant p < 0.05 compared to standard drug, Imipramine

Mechanism: Antidepressant (Serotonergic/histaminergic receptor
involvement)
Cyproheptadine (0.5 mg/kg, i.p.) on
reduction of immobility period
induced by ADE (30 mg/kg, p.o.) in
mice on TST (Panel A) and FST
(Panel B) models
• VEH: Vehicle (Normal saline, 10
ml/kg, p.o.);
i.p.);
71
Panel A
0
50
100
150
200
250
Immobility
time
(sec)/6
min
# #
Panel B
0
20
40
60
80
100
120
140
160
180
Immobility
time
(sec)/6
min
# #
ADE STR

Mechanism: Antidepressant (Serotonergic/histaminergic receptor
involvement)
Cyproheptadine (0.5 mg/kg, i.p.) on
reduction of immobility period
iduced by EF (250 mg/kg, p.o.) on
TST (Panel A) and FST (Panel B)
models
difformis rhizome (250 mg/kg, p.o.);
i.p.);
72
Panel A
0
50
100
150
200
250
Immobility
time
(sec)/6
min
#
#
Panel B
0
20
40
60
80
100
120
140
160
180
Immobility
time
(sec)/6
min
#
EF STR
#

Antipsychotic Test: Swim-induced grooming (SIG)
73
Panel A
0
5
10
15
20
Frequency
of
grooming
*
*
*
*
#
#
#
#
#
#
Panel B
0
50
100
150
200
VEH 30 60 125 250 500 1000 HAL
Duration
of
grooming/5
min
ADE (mg/kg, p.o.)
*
*
*
#
#
#
#
#
#
Fig. 82: Effects of ADE on frequency (Panel A) and duration (Panel B) of SIG
behaviour in mice
ADE: Aqueous extract of A. difformis rhizome; VEH: Vehicle-treated control group
(Normal saline, 10 ml/kg, p.o.); HAL: Haloperidol (1 mg/kg, i.p.); *significant p < 0.05
compared to control; #significant p < 0.05 compared to the reference drug, Haloperidol.

74
Panel A
0
2
4
6
8
10
12
14
Frequency
of
grooming/5
min
*
*
*
* *
*
#
#
#
#
#
# *
Panel B
0
50
100
150
200
250
300
VEH 30 60 125 250 500 1000 HAL
Duration
of
grooming
/5
min
HF (mg/kg, p.o.)
*
*
#
# #
# # #
Fig. 84: Effects of HF on frequency (Panel A) and duration (Panel B) of SIG behaviour
in mice
HF: N-Hexane fraction of A. difformis rhizome; HAL: Haloperidol (1 mg/kg, i.p.); VEH:
Vehicle (2.5% Tween 80 in saline, 10 ml/kg, p.o.); *significant p < 0.05 compared to vehicle-
treated control; #significant p < 0.05 compared to Haloperidol (1 mg/kg, i.p.)

75
Panel A
0
2
4
6
8
10
12
14
Frequency
of
grooming
/5
min
*
*
*
* *
*
*
#
#
# # #
#
Panel B
0
100
200
300
VEH 30 60 125 250 500 1000 HAL
Duration
of
grooming/5
min
EF (mg/kg, p.o.)
*
*
#
Fig. 85: Effects of orally administered EF on frequency (Panel A) and duration (Panel
B) of SIG behaviour in mice
EF: Ethyl-acetate fraction of A. difformis rhizome; HAL: Haloperidol (1 mg/kg, i.p.); VEH:
Vehicle (2.5% Tween 80 in saline, 10 ml/kg, p.o.). *significant p < 0.05 compared to
vehicle-treated control; #significant p < 0.05 compared to Haloperidol (1 mg/kg, i.p.).

• Fig. 86: Effects of BF on
frequency (Panel A) and
duration (Panel B) of SIG
behaviour in mice
difformis rhizome;
• HAL: Haloperidol (1 mg/kg,
i.p.);
• VEH: Vehicle (2.5% Tween 80
in saline, 10 ml/kg, p.o.);
to vehicle-treated control;
to Haloperidol (1 mg/kg, i.p.).
76
Panel A
0
2
4
6
8
10
12
14
Frequency
of
grooming
/5
min
*
*
*
* * *
*
#
#
#
# # #
Panel B
0
50
100
150
200
250
300
VEH 30 60 125 250 500 1000 HAL
Duration
of
grooming/5
min
BF (mg/kg, p.o.)
*
# # # # #
#

• Fig. 87: Effects of AF on
frequency (Panel A) and
duration (Panel B) of SIG
behaviour in mice
difformis rhizome;
• VEH: Vehicle-treated control
group (Normal saline, 10
ml/kg, p.o.);
• HAL: Haloperidol (1 mg/kg,
i.p.); *significant p < 0.05
compared to control;
to the reference drug,
Haloperidol
77
0
2
4
6
8
10
12
14
16
Frequency
of
grooming
/5
min
*
*
* *
* *
*
#
#
# #
#
#
Panel A
Panel B
0
50
100
150
200
250
VEH 30 60 125 250 500 1000 HAL
Duration
of
grooming/5
min
AF (mg/kg, p.o.)
*
*
*
*
#
# #
#
#
#

Conclusion
• The extract and fractions of A. difformis in this study demonstrated
a central nervous system depressant activity at high doses where
they exerted a sedative effect which was comparable to Diazepam.
• They also showed anxiolytic and antidepressant properties at low
doses in mice.
• The sedative effect appeared to be exerted via GABAA receptor;
• The anxiolytic effect seemed to be exerted via GABAAergic and
serotonergic/histaminergic pathways;
• The antidepressant activity was probably exerted via adrenergic,
serotonergic/histaminergic pathways;
• These findings provide a scientific basis for the ethno-medicinal
use of the rhizome of A. difformis in the management of mental
illness in the South-western part of Nigeria.
78

References
• Agyare C, Boakye YD, Apenteng JA, Dapaah SO, Appiah T, Adow A (2016)
Antimicrobial and anti- inflammatory properties of Anchomanes
difformis (Bl.) Engl. and Colocasia esculenta (L.) Schott. Biochemistry
and Pharmacology 5: 201- 204.
• Akah PA and Njike HA (1990) Some pharmacological effects of rhizome
aqueous extract of Anchomanes difformis. Fitoterapia; 61: 368-370.
• Akanmu MA, Olowookere TA, Atunwa SA, Ibrahim OI, Lamidi OF, Adams PA,
Ajimuda BO, and Adeyemo LE (2011)
Neuropharmacological effects of Nigerian honey in mice. African
Journal of Traditional Complementary and Alternative
Medicine; 8 (3): 230-249.
• Aliyu AB, Ibrahim MA, Musa AM, Musa AO, Kiplimo JJ, Oyewale AO (2013)
Free radical scavenging and total antioxidant capacity of
root extracts of Anchomanes difformis Engl. (Araceae). Acta Poloniae
Pharmacuetica-Drug Research; 70 (1): 115 – 121.
• Atawodi SE, Bulus T, Ibrahim S, Ameh DA, Nok AT, Mamman M, Galadima M
(2003) In vitro trypanocidal effect of methanolic extract of
some Nigerian savannah plants (abstract). African Journal of
Biotechnology; 2 (9): 317.
• Eke IG, Obioha FC, Anaga AO (2013) Evaluation of the Methanolic Rhizome
Extract of Anchomanes difformis for Analgesic and Antipyretic Activities.
International Journal of Basic and Applied Sciences; 2 (4): 289-296.
• Eneojo AS, Egwari LO, Mosaku TO (2011) In vitro Antimicrobial Screening on Anchomanes difformis
(Blume) Engl. Leaves and Rhizomes Against Selected Pathogens of Public Importance. Advances in
Biological Research 5 (4): 221-225.
79

References
• File SE and Pellow S (1985) The effect of triazolobenzodiazepines in two animal tests of
anxiety and on the hole-board. British Journal of Pharmacology 86: 729–
735.
• Lorke D (1983) A new approach to practical acute toxicity testing. Archieves of Toxicology;
54: 275 – 287.
• Magaji MG, Yakubu Y, Magaji RA, Musa AM, Yaro AH, Hussaini IM (2014)
Psychopharmacological Potentials of Methanol Leaf Extract of Securinega
virosa Roxb (Ex Wild) Baill. in Mice. Pakistan Journal of Biological Sciences;
17: 855 – 859.
• Nic Dhonnchadha BA, Bourin M., Hascoet M (2003) Anxiolytic-like effects of 5-HT2
ligands on three mouse models of anxiety. Behavioral Brain Research, 140:
203-214.
• Olayiwola G, Ukponmwan O, Olawode D (2013) Sedative and anxiolytic effects of the
extract of the leaf of Stachytarphyta cayennensis in mice. African Journal of
Traditional Complementary and Alternative Medicine; 10(6): 568-579
• Porsolt RD, Bertin A, Jalfre M (1977) Behavioral despair in mice: a primary screening test
for antidepressants. Archives of International Pharmacodynamics and
Therapeutics; 229: 327–336.
• Simiand J, Keane PE, Moore M (1984) The staircase test in mice: a simple and procedure
for screening of anxiolytic agents. Journal of Psychopharmacology
(Berlin) 84: 48– 53.
• Steru L, Chermat R, Thierry B and Simon P (1985) The tail suspension test: a new method
for screening antidepressant drugs. Psychopharmacology; 85:367–370.
• Trullas R, Skolnick P (1993) Differences in fear motivated behaviors among inbred mouse
strains. Psychopharmacology (Berl); 111(3): 323-31.
80

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