J. Afr. Ass. Physiol. Sci.

8 (2): 149-157, December 2020

Journal of African Association of Physiological Sciences

Official Publication of the African Association of Physiological Sciences
http://www.jaaps.aapsnet.org

Research Article
Anticonvulsant, antiamnesic and anxiolytic activities of methanol
leaf extract of Bambusa vulgaris (Poaceae) in mice

M.A Adebayo1, L.A Akinpelu2*, E.O. Okwuofu2, D.E Ibia1, A.F Lawson-Jack2, I.
Igbe2,3
Departments of 1Pharmacognosy and 2Pharmacology and Toxicology, College of Pharmacy, Igbinedion
University, Okada, Edo State, Nigeria and 3Pharmacology and Toxicology, Faculty of Pharmacy, University of
Benin, Benin City, Edo State, Nigeria
Keywords: ABSTRACT
Bambusa vulgaris, Background: Previous findings have shown that epilepsy can precipitate amnesia and anxiety,
anticonvulsant, among other neuropsychiatric disorders. Bambusa vulgaris is used in African traditional
antiamnesic, anxiolytic, medicine against convulsion, amnesia and anxiety but there is scanty scientific basis for these
tannin, Fourier ethnomedicinal claims. Hence, this study investigated the anticonvulsant, antiamnesic and anti-
Transform-Infra Red anxiety effects of Bambusa vulgaris in mice. Methods: The acute oral ingestion of Bambusa
spectra vulgaris (100, 200 and 400 mg/kg) was investigated using pentylenetetrazole-, and strychnine-
induced convulsion; antiamnesic using scopolamine-, and diazepam-induced amnesic models
while the anxiolytic effect was assessed using elevated plus maze models. The phytochemical
analysis was carried out using standard methods. Results: The extract at all the doses used
significantly (p<0.05) elongated the death latency while at 400 mg/kg the onset of clonic and
tonic convulsions were significantly (p<0.05) prolonged in pentylenetetrazole-induced
convulsion model. The extract at 100, 200 and 400 mg/kg offered 60, 80 and 100% protection
respectively in pentylenetetrazole-induced convulsion test. The extract showed no significant
(p>0.05) effect on strychnine-induced convulsion model ruling out the involvement of
strychnine-sensitive glycine receptor in the anticonvulsant effect of the extract. The extract at all
the tested doses significantly (p<0.05) in a dose dependent fashion ameliorated the amnesia
induced by scopolamine and diazepam suggesting antiamnesic effect. Bambusa vulgaris at all
the tested doses significantly (p<0.05) in a dose dependent pattern increased the percentage open
arm entries and percentage open arm duration on the open arm of the elevated plus maze as well
as reduced the anxiety indices of the experimental mice consistent with anxiolytic effect. The
phytochemical quantification of the extract showed abundance of tannins and corroborated by
the findings from the Fourier transform infrared spectra of the extract. Conclusion: This study
therefore concluded that Bambusa vulgaris may possess anticonvulsant, antiamnesic and
anxiolytic effects and provided scientific proof for its traditional use.
© Copyright 2020 African Association of Physiological Sciences -ISSN: 2315-9987. All rights reserved

INTRODUCTION
excitatory signals in the brain (Kolawole et al., 2012).
Epilepsy according to the World Health Organisation is
Unfortunately, these antiepileptic drugs can only abolish
a chronic non-communicable brain disorder
seizure in nearly 70% of the patients suffering from
characterized by seizure, which affect individuals of all
epilepsy without complete remission in the remaining
age brackets (WHO, 2012; Kolawole et al., 2012). The
30% of the epileptic patients (Sander, 2004).
currently available antiepileptic drugs target
Earlier research investigation has indicated that epilepsy
GABAergic, serotonergic, noradrenergic and
can precipitate amnesia (Gallassi, 2006) and anxiety
dopaminergic systems involved in inhibitory and
among other neuropsychiatric disorders (Uruaka and
Georgwill, 2020).
Anxiety has earlier been described as “a psychological,
*Address for correspondence: physiological, and behavioral state induced in animals
Email: [email protected] and humans by a threat to well-being or survival, either
 Anticonvulsant, antiamnesic and anxiolytic activities of Bambusa vulgaris

actual or potential.” (Steimer, 2002). It is associated with collection was done within the temporary site of the
a wide range of psychiatric conditions such as epilepsy Igbninedion University Campus Okada in Ovia North
and neurodegenerative disorders (Stein et al., 1990). The Eat Local Government of Edo State. The leaves were
major classes of drugs employed in the treatment of authenticated by Mr. G. A. Ademoriyo of the Herbarium
anxiety disorder are benzodiazepines and selective Unit, Department of Botany, Faculty of Sciences,
serotonin-reuptake inhibitors (SSRIs) (Parek and Obafemi Awolowo University, Ile-Ife and herbarium
Chanda, 2006), but the undesirable effects linked with number IFE-17960 was obtained.
these classes of drugs (Kunovac and Stahl, 1995) have
unfortunately limited their use. Extraction of plant material
The undesirable effect associated with the synthetic The extraction protocols used in this study was as earlier
anxiolytic drugs (Kunovac and Stahl, 1995) and the reported (Velavan, 2015) with mild modification.
inability of the currently available antiepileptic drugs to Briefly, fresh leaves of Bambusa vulgaris were
control epileptic seizure (Stein et al., 1990), coupled collected, shade dried and subsequently ground into
with the untoward effects linked to acetylcholineesterase powder with mechanical grinder. The powdered material
inhibitors used against amnesia (Baradaran et al., 2012) was macerated in 70% methanol for 72 h. The resulting
have undoubtedly warranted the search for novel filtrate was concentrated in vacuo and subsequently
anticonvulsant, antiamnesic and anxiolytic medications freeze dried to yield 12.6 g (4.2 %) coded BV which was
probably from medicinal plants. stored in a desiccator prior to use. The extract was
Bambusa vulgaris, commonly referred to as “Bamboo” dissolved with 3% Tween 80 and made up to the
(Carey et al., 2009) is a member of the Poaceae family. required volume with normal saline and prepared fresh
Bamboo is a large perennial grass dispersed widely from on each day of the experiment.
tropical to subarctic zones (Ambika and Rajagopal,
2019) and commonly found in moist places such as the Experimental animals
river banks (Khan and Hemalatha, 2015). Adult male mice of between 18-25 g were purchased
The different parts of bamboo such as leaf, shoot, stem from the Central Animal House of the Igbinedion
and sap are used as medicine in folkloric medicine. In University Okada. The animals were fed with standard
Nigerian folkloric medicine, bamboo is reported to be animal pellets with water ad libitum. The experimental
used in the management of gonorrhea, respiratory guidelines followed in this research investigation were
diseases, as arbotifacient, as well as an emmenagogue as approved and being implemented by the Igbinedion
and appetizer (Yakubu and Bukoye, 2009). It is also used University Animal Ethical Committee which are in line
in Nigerian traditional medicine as anti-aging and with the internationally accepted principles for
memory enhancer (Elufioye et al., 2012). In Indian Laboratory Animal Use and Care (Waldegrave, 1986).
folkloric medicine, the leaves are used in the treatment
of various inflammatory conditions (Carey et al., 2009). Drugs
In Chinese ethnomedicine, bamboo is used to ease labor Diazepam (DZP) (Roche, Basel, Switzerland);
and placenta expulsion (Ambika and Rajagopal, 2019). Pentylenetetrazol (PTZ), Strychnine (SCN),
The anti-inflammatory (Carey et al., 2009), antioxidant Scopolamine hydrobromide (SCOP), Piracetam
(Goyal et al., 2013), arbotifacient (Yakubu and Bukoye, (PIRAC), Tween 20 (Sigma Chemicals Co, St. Louis,
2009), antimicrobial and antiproliferative (Ambika and Missouri, U.S.A.); Phenobarbitone (May and Baker,
Rajagopal, 2019) activities of Bambusa vulgaris have United Kingdom); and normal saline (Unique
been reported. Pharmaceutical Limited, Lagos, Nigeria).
This study aimed to investigate the anticonvulsant,
antiamnesia and anxiolytic potentials of Bambusa Spectrophotometric phytochemical estimation
vulgaris in mice upon its use in traditional medicine as The tannin content, total flavonoids, total phenols and
anticonvulsant, memory enhancer and anxiolytic agent total alkaloids in BV were estimated as earlier reported
(Clark et al., 1995; Ogunjimi et al., 2009; Elufioye et al., in literature (Akinpelu et al., 2019a).
2012).
Spectroscopic analyses of BV
MATERIALS AND METHODS The extract was scanned at the wavelength range of 200-
Plant identification, collection and authentication 900 nm to obtain the UV-VIS spectrum of BV using UV
Fresh leaves of Bambusa vulgaris were collected by Mr spectrophotometer. The FT-IR spectrum of BV was
M.A Adebayo of the Department of Pharmacognosy, obtained after mixing the extract with the spectra-grade
College of Pharmacy, Igbinedion University Okada. The potassium bromide (KBr) in the ratio of 1:100 and

150 J. Afr. Ass. Physiol. Sci. 8 (2): 2020 Adebayo et al.

 Anticonvulsant, antiamnesic and anxiolytic activities of Bambusa vulgaris

pressed into pellet. The pellet was immediately number of triads was recorded as ‘percentage
introduced into the sample holder of Perkin Elmer alternation’ which indicates the assessment of short-term
Spectrophotometer. The Spectrophotometer was memory (Sarter et al., 1988; Akinpelu et al., 2020),
operated in the range 4000 – 300 cm–1 to detect the according to this equation: An alternation is defined as
spectra data of the different functional groups present in an entry into all three arms on consecutive choices%
the extract (Akinpelu et al., 2019a). Alternation = [(Number of alternations)/ (Total arm
entries -2)] x 100.
General experimental design
Adult male albino mice were randomly sorted into five Diazepam-induced amnesia
experimental groups (n=5). Group-I (negative control) The experiment was carried out as above for
mice received 3% Tween 80 in normal saline (10 mL/kg, scopolamine-induced amnesia, but diazepam (1 mg/kg,
p.o.). Group II-IV mice received BV (100, 200 or 400 i.p.) was administered to induce amnesia instead of
mg/kg, p.o.), while Group-V received intraperitoneal scopolamine (Akinpelu et al., 2020).
injection of positive control drug such as piracetam (200
mg/kg, p.o.) for scopolamine and diazepam-induced Anxiolytic test
amnesia; diazepam (1 mg/kg, i.p.) for anxiolytic and Elevated plus maze (EPM):
PTZ-induced convulsion experiments, while The test was conducted as recently carried out and
phenobarbitone (30 mg/kg, i.p.) was used as positive reported in literature (Akinpelu et al., 2019a). After the
control drug in strychnine-induced convulsion model. respective oral ingestion with BV or intraperitoneal
administrations with piracetam, each mouse was singly
Anticonvulsant tests placed in the central square of the EPM facing one of the
Pentylenetetrazole (PTZ)- induced open arms and the number of entries into the open arm
One hour post oral ingestion of BV (100, 200 and 400 and the time spent on the open arm were noted with a
mg/kg) or half an hour post intraperitoneal treatment stopwatch and recorded manually for a period of 5
with diazepam (1 mg/kg), mice in all groups I-V mice minutes.
were injected with the chemoconvulsant agent PTZ (85 The open arm avoidance index interpreted as level of
mg/kg, i.p.). Immediately after the intraperitoneal anxiety (Trullas and Skolnick, 1993) is calculated as
injection of PTZ, each mouse was gently put in an
observation cage and observed for 30 minutes duration
for the onset of clonic, tonic convulsions and death time
which were recorded with cut off time of 30 minutes and Statistical analysis
the mice that survived beyond 30 minutes were Results are expressed as mean ± S.E.M and analysed
considered protected (Swinyard et al., 1989; Akinpelu et using one-way analysis of variance (ANOVA), followed
al., 2018). by Dunnett’s post hoc analysis. GraphPad InStat®
Biostatistics software (GraphPad Software, Inc., La
Jolla, USA) was employed as the statistical tool. The
Strychnine (SCN)-induced convulsion
level of significance for all tests was set at p <0.05
The experiment was carried out as above for PTZ-
compared to the control group.
induced convulsion except that strychnine (4 mg/kg, i.p.)
was used as chemoconvulsant (Swinyard et al., 1989)
and phenobarbitone (30 mg/kg, i.p.) as positive control RESULTS
drug in this model (Akinpelu et al., 2018). Effect of Bambusa vulgaris on PTZ-induced convulsion
model
Antiamnesic tests The extract at 100 and 200 mg/kg showed significant
(p<0.05) prolongation of death time and offered 60 and
Scopolamine-induced amnesia
80% protection respectively compared to the control
Thirty minutes after oral ingestion of normal saline (10
which offered 0% protection. The extract at 400 mg/kg
mL/kg) in group I, BV (100, 200 and 400 mg/kg, p.o.) in
significantly (p<0.05) elongated the onset of clonic,
groups II-IV and piracetam (200 mg/kg, p.o.) in group
tonic convulsions, death latency and offered 100%
V, scopolamine (1 mg/kg) was intraperitoneally injected
protection. The result is presented in Table 1.
to each mouse in groups I-V, thirty minutes after
scopolamine injection, each mouse was observed on Y-
Effect of Bambusa vulgaris on SCN-induced convulsion
maze (40 x 3 x 12) with angles of 1200 between each of
model
the three arms and the sequence of arms visited on
The extract at all the doses used elongated the onset of
consecutive choices in 8 minutes was recorded. The
clonic, tonic convulsions and death time but not
151 J. Afr. Ass. Physiol. Sci. 8 (2): 2020 Adebayo et al.
 Anticonvulsant, antiamnesic and anxiolytic activities of Bambusa vulgaris

Table 1: Effect of BV on PTZ-, and SCN-induced Effects of BV on percentage alternation in scopolamine-

convulsions in mice. induced amnesia on Y-Maze task.
Scopolamine significantly (p<0.05) lowered the
percentage correct alternation on Y-maze when
compared to the control treated control group. However,
BV significantly (p<0.05) and in a dose dose dependent
pattern increased the reduced alternation induced by
Scopolamine when compared to the Scopolamine treated
control group. Piracetam, a positive control drug
significantly (p<0.05) reversed the reduced alternation
induced by Scopolamine in mice. The result is presented
in Fig. 1 (Panel A).

Effects of BV on percentage alternation in Diazepam-

induced amnesia in Y-Maze task.
Values are Mean ± SEM, ANOVA; one-way analysis of variance
Diazepam showed significantly (p<0.05) reduction in
followed by Dunnett’s post hoc Test, n=5, *p<0.05 compared to
control (ANOVA; Dunnett’s post hoc test). the percentage correct alternation on Y-maze when
compared to the control treated control group. However,
significant (p>0.05) from the control treated mice. No BV showed significant (p<0.05) reversal in a dose
protection was as well offered at the the tested doses. dependent manner the reduced alternation induced by
However, Phenobarbitone (30 mg/kg, i.p., a positive Diazepam when compared to the Diazepam treated
control drug) significantly (p<0.05) prolonged the onset control group. Piracetam, a positive control drug
of clonic, tonic convulsions, death latency and offered significantly (p<0.05) attenuated the reduced alternation
50% protection. The result is presented in Table 1.

Fig. 1: Effect of Bambusa vulgaris on percentage alternation in scopolamine-induced amnesia (Panel A) and diazepam-induced amnesia
(Panel B) in mice. Each bar represents Mean ± SEM, n=5. #p<0.05 and *p<0.05 compared to the control and scopolamine scopolamine
(Panel A) or diazepam (Panel B) treated mice respectively (ANOVA; Dunnett’s post hoc test).

induced by Diazepam in mice. The result is presented in control mice. The effect of the extract at 400 mg/kg is
Fig. 1 (Panel B). comparable to diazepam (1 mg/kg, i.p., a positive control
drug). The result is presented in Fig. 2 (Panel A).
Effect of Bambusa vulgaris on percentage open arm
entries Effect of Bambusa vulgaris on percentage open arm
The extract significantly (p<0.05) and dose dependently duration
increased the percentage number of entries into the open The extract of Bambusa vulgaris significantly (p<0.05)
arm of the elevated plus maze when compared to the and dose dependently increased the percentage time

152 J. Afr. Ass. Physiol. Sci. 8 (2): 2020 Adebayo et al.

 Anticonvulsant, antiamnesic and anxiolytic activities of Bambusa vulgaris

Fig. 2: Effect of Bambusa vulgaris on percentage open arm entries (Panel A) and Percentage open arm duration (Panel B) on
elevated plus maze. Each bar represents Mean ± SEM, n=5. *p<0.05 compared to the control treated mice.

Table 2: Quantitative phytochemical analysis of

methanol leaf extract of Bambusa vulgaris

Values are means of triplicate determination ± Standard

deviation; where AE is atropine equivalent, GAE is gallic acid
Fig. 3: Effect of Bambusa vulgaris on anxiety index on
equivalent, QE is quercetin equivalent and AE is atropine
elevated plus maze. Each bar represents Mean ± SEM, n=5. If
equivalent respectively.
the anxiety index is at least 10 point less than the vehicle
treated control group, the sample is anxiolytic, conversely if
spent on open arms of the elevated plus maze when the anxiety index is at least 10 points greater than the vehicle
compared to the control mice. The effect of the extract treated control group, then the sample is anxiogenic.
at 400 mg/kg is comparable to diazepam (1 mg/kg, i.p.,
a positive control drug). The result is presented in Fig. 2 Result of quantitative phytochemical quantification of
(Panel B). Bambusa vulgaris
The result showed that Tannin content > total
Effect of Bambusa vulgaris on open arm avoidance flavonoids > total phenols > total alkaloids. The result
index (OAAI) is presented in Table 3.
The extract showed at least a ten-point reduction in
anxiety index of open arm avoidance index in a dose UV-VIS Spectrum Peak values of BV
dependent manner when compared to the control treated The UV-VIS spectrum of BV showed absorption peaks
mice. The effect of the extract at 400 mg/kg is at 209.0, 296.5, 349.0, 490.0, 496.5, 556.5, 604.0, 663.5,
comparable to diazepam (1 mg/kg, i.p., a positive control 680.5, 691.0, 718.0, 725.0, 732.5, 752.5, 760.5, 766.5,
drug). The result is resented in Fig. 3.

153 J. Afr. Ass. Physiol. Sci. 8 (2): 2020 Adebayo et al.

 Anticonvulsant, antiamnesic and anxiolytic activities of Bambusa vulgaris

Table 3: FTIR profile of methanol leaf extract of characteristic bands at 3302.4, 2944.6, 2832.8, 1636.3,
Bambusa vulgaris 1449.9, 1397.8, 1338.1, 1110.7, 1017.6 and 641.1 cm-1.
The result is presented in Table 4.

DISCUSSION
The present investigation assessed the anxiolytic and
anticonvulsant potentials of methanol leaf extract of
Bambusa vulgaris and the probable phytocompounds
responsible for these pharmacological effects. The
extract demonstrated anticonvulsant, antiamnesic and
anxiolytic effects with tannins being the most abundant
phytoconstituents assayed.
The prolongation of the onset of clonic, tonic
convulsions and death latency coupled with the varying
degree of protection at the tested doses in PTZ-induced
convulsion model suggest that the extract may possess
anticonvulsant effect. This finding is in consonance with
earlier suggestion of medicinal plant agent that showed
797.5, 837.5, 849.0, 863.0 and 888.5 nm and the anticonvulsant effect in PTZ-induced convulsion in mice
absorption of 3.776, 0.681, 0.463, 0.027, 0.021, 0.010, (Akinpelu et al., 2018).
0.005, 0.010, 0.010, 0.003, 0.002, 0.002, 0.001, 0.005, The anticonvulsant effect of the extract may be acting
0.007, 0.004, 0.003, 0.003, 0.005, 0.006 and 0.004 via GABAA-benzodiazepine receptor neurotransmission
respectively (Fig. 4). since GABAergic pathway is key to the anticonvulsant
FTIR spectrum of BV potential of medicinal agents in PTZ induced convulsion
Table 2 revealed the FTIR spectrum of BV which showed models (Akinpelu et al., 2018).

Fig. 4 (Left): UV-VIS spectrum profile of methanol leaf extract of Bambusa vulgaris
Fig. 5 (Right): FTIR spectrum profile of methanol leaf extract of Bambusa vulgaris

The amelioration of the amnesia induced by central muscarinic cholinergic receptor, while diazepam
scopolamine and diazepam by the extract suggests that induces memory deficit either via acting as either agonist
the extract may have antiamnesic effect probably acting or antagonist at the central GABAA-benzodiazepine
via central muscarinic cholinergic and/or GABAA-BDZ receptor pathway (GABAA-BDZ) since GABAA-BDZ
receptor neurotransmission to elicit the observed receptor complex have been earlier reported to control
antiamnesic effects. Since scopolamine induces memory the release of acetylcholine (Vazquez and Baghdoyan,
impairment via acting as an antagonist action at the 2003), a neurotransmitter pivotal to learning and
central muscarinic cholinergic receptor, while diazepam
induces memory deficit either via acting as either agonist
154 J. Afr. Ass. Physiol. Sci. 8 (2): 2020 Adebayo et al.
 Anticonvulsant, antiamnesic and anxiolytic activities of Bambusa vulgaris

memory (Halsemo et al., 2006). The finding of this study stretch vibration of benzene nucleus and methylol group
is in accordance to the earlier reports of medicinal plants of tannin (Rajeswari and Jothiprakasam, 2017). Moreso
with antiamnesic effects in mice (Akinpelu et al., 2019a; that BV contained functional groups such as O-H, C-H,
2020). C=O, C=O-C aromatic and C-O earlier shown to be
The increase in number of open arm entries and time present in tannins (Grasel et al., 2015). These functional
spent on the elevated plus maze suggest that the extract groups could further support the abundance of tannins
may have anxiolytic effect since increase in these and other phenolic compounds quantitatively assayed in
parameters by medicinal plant has been attributed to an the extract.
anxiolytic effect in previous report (Akinpelu et al.,
2019a). Although it was not part of the objective of this CONCLUSION
study, but it is of interest to note that methanol leaf This study concluded that Bambusa vulgaris may
extract of Bambusa vulgaris extracted with soxhlet possess anxiolytic and anticonvulsant effect. This study
extraction showed anti-anxiety effect at the higher further concluded that tannin either in additive or
intraperitoneal dose of 400 mg/kg in mice (Kaur et al., synergy with other plant secondary metabolites may be
2019), but our study of the methanol leaf extract responsible for the observed pharmacological potentials.
extracted with cold maceration showed anti-anxiety
effect at the three tested doses of 100, 200 and 400 mg/kg
ACKNOWLEDGEMENTS
via oral route in mice
Authors are grateful to Mr. Ademoriyo G.A. of the
The extract may be exerting its anxiolytic effect via
department of Botany of the Obafemi Awolowo
GABAA-benzodiazepine receptor neurotransmission
University Ile-Ife for authenticating the plant and Mr
since elevated plus maze has been reported to be
Akpan Nya of the Central Animal House of the
sensitive to agents acting via this pathway (Nutt and
Igbinedion University, Okada for his assistance.
Maizia, 2001).
The qualitative phytochemical screening of medicinal
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