Evaluation of Anti-Epileptic Activity of Ethanolic Extract On Swiss Albino Male Mice

World Journal of Pharmaceutical Research
Divyashree et al. SJIF Impact

World Journal of Pharmaceutical Factor 8.084
Research
Volume 12, Issue 15, 556-591. Research Article ISSN 2277–7105
EVALUATION OF ANTI-EPILEPTIC ACTIVITY OF ETHANOLIC

EXTRACT ON SWISS ALBINO MALE MICE
*Divyashree L. M. Pharm. and Nagalakshmi N. C.M. Pharm.
Department of Pharmacology, Mallige College of Pharmacy, #71, Silvepura, Bangalore 90.
ABSTRACT
Article Received on
06 July 2023, Objective: To study the antiepileptic activities,ethanolic extract of the
Revised on 06 August 2023, leaves of Costus Igneus (EECI) in mice. Materials and Method: The
Accepted on 26 August 2023
DOI: 10.20959/wjpr202315-29190
antiepileptic activities of EECI was studied against maximal
electroshock (MES) and chemical (Pentylenetetrazole and strychnine)
induced seizure in mice. Result: The duration of tonic hind leg
*Corresponding Author
extension in MES model was significantly reduced by EECI at the
Divyashree L.
dose of 100mg/kg and 200mg/kg. In PTZ model, oral administration of
Department of Pharmacolo-
gy, Mallige College of EECI at dose of 200mg/kg significantly delayed the onset of clonic
Pharmacy, #71, Silvepura, convulsion and decrease the percentage of mortality. In strychnine
Bangalore 90. mode EERA at the dose of 100mg/kg and 200mg/kg significantly
delayed the onset of tonic extensior convulsion. Conclusion: The study
revealed the protective action of the EECI against the various model of epilepsy in
exprimental mice. This beneficial effect of Costus Igneus plant might be due to presence of
antioxidant (β-carotene), Saponins, Flavonoids property and CNS depressent property.
KEYWORDS: Epilepsy, Costus Igneus, Maximalelectroshock, Pentylenetetrazole,

Strychnine seizure.
1. INTRODUCTION
Epilepsy is a common neurological disorder characterized by paroxysmal dysrhythmia, sei-
zure, with or without body convulsion and sensory of psychiatric phenomena.[1] Epilepsy is
the variety of disorder reflecting underlying brain dysfunction.[2] Epilepsy is the second most
common chronic neurological condition seen by neurologist after stroke.[3]
As proposed by the International Bureau for Epilepsy (IBE) and League Against Epilepsy
(ILAE), epilepsy is defined as a brain disorder characterized by an enduring predisposition to
www.wjpr.net │ Vol 12, Issue 15, 2023. │ ISO 9001:2015 Certified Journal │ 556
Divyashree et al. World Journal of Pharmaceutical Research
generate epileptic seizure and by the neurobiologic, congnitive, psychologicand social conse-
quences of this condition.[4]
During epilepsy nerve cell in the brain becomes disrupt causing seizure or period of unusual
behavior, sensation and sometime loss of consciousness. Seizure symptoms can vary wildly.
Some people simply stare blankly for few second during a seizure, while others repeatedly
twitch their arm or leg.[5]
Seizure episodes are result of excessive electrical discharge in the group of brain cells and
can affect the any part of the brain. Frequency may vary from 1 per year to several times per
day. One seizure doesn’t signify epilepsy because up to 10% of the worldwide have one sei-
zure during their life period.[6]
Epilepsy is the Greek word as “epilambanein” meaning “to be seized” or “to be overwhelmed
by surprise”.[7] Epilepsy usually begins in children, potentially impeding education, employ-
ment, social relationship and development of a sense of self-worth.[8]
1.1. Incidence and prevalence of epilepsy

It is the most common neurological disorder with reported prevalence and incidence 5 per
1000 in India and slightly higher in rural area. The high prevalence is found in adolescent and
early age group. In North, Central and South America high prevalence is found in male ex-
cept in New York were as Bolivia, Honduras and Argentina female has high prevalence. In
Asian country such as China, India, Turkey and Saudi Arabia, the prevalence is high in male
except in Pakistan where prevalence is high in female. In European country prevalence is
high in female.
The incidence is higher in developing country than the industrialized country. Similarly inci-
dence is higher Male then Female.
Generally it is found that there is increase prevalence in adolescence and early childhood age.
Adolescent and childhood are commonly idiopathic (primary) in nature whereas rests of the
groups are commonly secondary epilepsy and in case of gender man has higher prevalence
then female.
1.2. History of epilepsy[9-11]

Being the word epilepsy is derived from the Greek and Latin word for “seizure” or “to seize
upon” which implies that epilepsy is an ancient disorder. British Museum, London is the ear-
liest detailed account of epilepsy which was written over 3000 years ago, i.e. before 1000
BC. The Babylonians were keen observer of clinical phenomena and provide remarkable de-
scription of many of the seizure types that we recognize today viz. tonic clonic, absence, drop
attack, simple and complex partial seizure and even focal motor or gelastic attacks. They
don’t know the pathophysiology but they say that it is because of evil spirit so treatment was
not medical but spiritual. They believed that the diseases was also caused by the presence of
the demons in the person and believe that if the person breathed on or touched another, the
demon would spread to the other person unless that person spit immediately.
Unsuccessful challenged by the School of Hippocrates in 5th century BC Greece, which was
first suggested that the brain was the seat of this disorder. Hippocrates said epilepsy as the
natural diseases and could be treated through natural product and method. He suggests the
use of medicine and control of the diet in order to cure the diseases. He had some notation
that epilepsy could become chronic and intractable if not treated effectively and early alt-
hough it is not clear what treatment he had in mind. Unfortunately the Hippocratic concept of
a treatable brain disorder had little influence.
In 17th and 18th century epilepsy was one of the several key area of debate in the gradual
identification and separation of “nerve disorder” from “mental disorder” which lead to begin-
ning of modern neurology of 19th century. Later it has been separated off from epilepsy in the
19th century by Robert Bentley Todd in 1849 and Jackson in 1890 by Lectures on convulsion
diseases.
Jackson concludes that motor cortex was concerned with movement rather than individual
muscles. By using Faradays concept of “disruptive discharge” Jackson viewed seizure as a
result of electric discharge in the brain, which he confirmed experimentally in the rabbit.
In 1929 Berger reported the discovery of the human EEG which leads rapidly conformation
of seizure were the result of electrical discharge in brain. In 1952 Hodgkin and Huxley win
the Nobel Prize on discovers of the ionic basic of Todd’s nervous polarity/force.
Modern therapy of the epilepsy probably began with bromides in 1856, Phenobarbital in
1912, and Phenytoin 1938. These are still using with great effectiveness.
1.3. Seizure type and clinical manifestation

Table 1: describes types of seizure and their clinical manifestations.
Table 1: Seizure type and there clinical manifestation.[12]
Seizure type Key Ictal EEG Manifestation Major clinical manifestation
Seizure may be limited to a limb or
muscle group; may show sequential
involvement of body parts; con-
I. Partial(focal, local) sei-
sciousness usually preserved; may be
zure Local contralateral discharge.
somatosensory; may have autonomic
 Simple partial seizure
symptom or sigh such as epigastric
sensation, sweeting, and papillary di-
lation.
Impairment of consciousness may
Unilateral or bilateral asyn-
have automatisms, flash back; auto-
 Complex partial seizure chronous focus, most often in
nomic activity such as pupil dilation,
temporal region.
flushing, piloeraction.
 Partial seizure evolving
May generalize to tonic, clonic, or
to secondary general-
tonic-clonic.
ized seizure
Brief loss of consciousness with or
II. Generalized seizure
with our motor involvement; occurs
 Absence seizure 3-Hz polyspike and wave.
in childhood with a tendency to dis-
(petit mal epilepsy)
appear following adolescence.
Sudden, brief, shock like contraction
 Myoclonic seizure
of musculature
Fast activity (10Hz or
more) increasing in ampli-
Fast activity (10 Hz or more;
tude during tonic phase; Repetitive muscle jerks.
slow waves).
interrupted by waves during
clonic phase.
Loss of consciousness; sudden sharp
tonic contraction of muscle, falling to
 Tonic- clonic seizure
ground, following by clonic convul-
(grand mal epilepsy)
sion movement; often postictal de-
pression and incontinence.
Rigid, violent muscular contraction
 Tonic seizure Low voltage, fast activity.
with limbs fixed.
Sudden diminution in muscle tone
affecting isolated muscle group or
 Atonic seizure (astatic) Polyspikes and wave.
loss of all muscles tone; may have
extremely brief loss of consciousness.
1.4. Causes of epilepsy

Epilepsy has no identifiable cause but condition may be traced to various factors:[13,14]
 Genetic influence
Some type of epilepsy run in families. It is estimated up to 500 genes could be tied to the
condition. Gene may make person more sensitive to environment condition that trigger sei-
zure.
 Head trauma
Head trauma as a result of car accident or other traumatic injury can cause injury.
 Brain conditions
Like brain tumour or strokes can cause epilepsy. Stroke is the leading cause of epilepsy in
adult older than age 35.
 Infectious diseases
Like AIDS, meningitis and viral encephalitis can cause epilepsy.
 Prenatal injury
Before the birth of the child, the brain of the child is more sensitive, any factor such as infec-
tion on mother, poor nutrition or oxygen deficiencies.
 Developmental disorder
Some time it associated with the disorder such as autism and neurofibromatosis.
 In people over 65, stroke is the most common cause of new onset seizures. Other condi-
tions such as Alzheimer’s disease or other conditions that affect brain function can also
cause seizures.
1.5. Pathophysiology of epilepsy[15-18]

Figure 1 shows pathophysiology of epilepsy. Neuronal message are transmitted by electrical
impulses called the action potential. This is actually a net positive inward ion flux that leads
to depolarization or voltage change in the neuronal membrane. The ions involved include so-
dium, potassium, calcium and chloride. In normal brain, hyper excitability is prevented by
inhibitory mechanism involving negative ions like chloride ions. Disturbance of normal ex-
citability leads to hyper- excitability causes the increase in excitatory transmission of impulse
and decrease inhibitory transmission. Once activated impulses flow via the neuronal circuits
along the axons of the nerve. An action potential travels down the axon to the terminal but-
tons and then releases neurotransmitters in the synaptic cleft. Glutamate is the primary excita-
tory neurotransmitter in the brain whereas GABA is the principal inhibitory neurotransmitter.
Glutamate are of two type, Ionotropic(NMDA receptor) that modulate calcium channel and
sodium channel and are responsible for fast synaptic transmission and Metabotropic (non
NMDA) that are for slow synaptic transmission. GABA is mediated via Chloride and Potas-
sium channel.
When any stimuli come in contact cause the inward of current of Na+, Ca++ and involves the
release of the neurotransmitter like Glutamate and Aspartate. Glutamate binds to the NMDA
receptor and causes the excitation of neuron. Patient with absence seizure, plasma glutamate
level were found to be significant increased. When there is diminish synthesis of GABA or
interaction of post synaptic GABA causes the convulsion. In the epileptic condition there is
decrease in the dopamine whereas nor-adrenalin increases.
The neuron of mortar cortex, limbs system and the temporal lobes are particularly epilepto-
genic.
Figure 1: Flow chart of pathophysiology of epilepsy.
1.6. Treatment of epilepsy[19-21]

1.6.1. Goal of treatment (desire outcome)
It is to control or reduce the frequency of the seizure, minimize the side effect and ensure
complains, allowing the patient to live a normal life.
1.6.2. General principle of management of epilepsy

In the some case treatment of cause can cure the seizure. It should be consider as illness not a
social stigma. Occupation like driving the vehicles, working on the machine near water, at the
height is not suitable. For the rational management of epilepsy it needs an accurate evaluation
of the epileptic syndrome. Currently there is no any medicine for cure epilepsy. So current
aim is to achieve prolong seizure free period with the low drug toxicity. Regular use of the
drug can abolish the seizure completely in 60-80% of the patient.
Vagus nerve stimulation is the non-pharmacological approaches for the treatment and man-
agement of epilepsy and is now widely used in treatment of partial seizure nowadays. This
therapy is for the patient who is poorly tolerated with anti -seizure drugs. Stimulatory elec-
trodes are kept on left vagus nerve and peace maker is implanted on the chest or axilla.
To get the best result with drugs following should be observe carefully:
 Proper initial evaluation is necessary.
 AED is advised if the patient is having two or more seizure, have the history of seizure,
abnormal neurological finding, abnormal EEG, and initial choice is depending on the type
of seizure.
 Therapy should be started with singe drugs and drug with less toxicity.
 Repeated examination of plasma and EEG helps in difficult case.
 Mild toxicity can be seen like skin rashes and can be manage by reducing dose of the
drugs.
 Attending regular follows up and for the children parents should ne council regarding
compliance and duration of treatment.
 Mon therapy with standard drugs can give satisfactory result but if it became inadequate
then another drug is substituted and again if it fails then combine therapy should be start-
ed.
 For second drug start first drugs should be tapped slowly and second drug should ne in-
troduce gradually.
 Patient with multiple seizure and focal epilepsy due to underlying structural lesion should
be treated with the multiple drugs and start with the older drugs and if resistance then new
drugs should be started.
 The patient should be advised to avoid using OTC formulation and drugs from alternative
medicine as it may contain drugs which lower seizure threshold and precipitate seizure.
 1.6.3. Pharmacological treatment of epilepsy[19-23]

 Table 2 depicts classification of AED based on mechanism of action.
Table 2: Classification of antiepileptic drug (AED) based in mechanism of action.
Class of drug Example
Decrease in the neuronal excitability by delaying the Phenytoin, Carbamazepine,
recovery of inactivated Na+ channel Lamotrigin, Topiramte
Decrease in the low threshold calcium channel cur-
Ethosuximide
rents(T- current) in the thalamic neuron
Action similar to both Phenytoin and Ethosuximide as
Sodium valproate, Zonisamiede
mention above
Enhancement of inhibition through GABA
Acting through GABA related receptor Barbiturate, Benaodiazepines
By releasing GABA from neuronal ending Gabapentin
By inhibiting GABA transaminase Sodium valproate, Vigabatrin
By inhibiting neuronal reuptake of GABA Tiagabine
Inhibition if excitatory neurotransmitter glutamate Lamotrigine
Miscellaneous Levetiracetam, Acetazolamide
1.6.3.1. Mechanism of action of different AED[24-27]

Generally the mechanism of action for the ADEs includes effect on the ion channel (sodium
and Ca++), inhibitory neurotransmission (increasing CNS GABA), or excitatory neurotrans-
mission (decrease or antagonizing glutamate and aspartate). AEDs that are effective against
general tonic clonic and partial seizures probably work by delaying recovery of sodium chan-
nel from activation. Drugs that reduce corticothalamic T- type Ca++ current are effective
against generalized absence seizure. Figure 2 deficits the different mechanism of action of
AED. Figure 3 and 4 show the structure of GABA receptor and mechanism of action of AED
binding on GABA receptor.
Figure 2: Representing the different mechanism of action of AED.
GABA receptor is the inhibitatory receprot and are of 3 types GABA-A, GABA–B and
GABA–C. GABA-A and GABA-C receptor is inotropic where as GABA-B is metabotropic
that modulate the oepning of the potassium channel throug secondary messenger involving a
G-protein. Amont all GABA-A is well known receptor and Benzodiazepine, Alcohol,
Barbiturate convulasnt steroid undergoes binding on it.
Figure 3: GABA receptor with different site for binding of drugs.
Figure 4: Mechanism of action of AED in GABA receptor.
1.6.3.2. Choice of AED

Initial in newly diagnosed patient first line AED (Table 3) should be given with the low dose
because of rapid introduction may leads to the side effect and also lose patient confidence.
For the maintenance dose there is no any AED that suits to all patients. Amount of the dose
varies from patient to patient and drugs to drugs. There should be gradual increment in the
dose and maintain. Control of the seizure should be assessed and the dose of the drugs should
be changed if necessary. More care is needed in Phenytoin as the serum level dose relation is
not linear because the small change in the dose can cause the considerable change in the se-
rum level. If the maximal tolerated dose of the drug doesn’t control seizure or if develop the
side effect then drug can be replaced with another first line drug. For initiation of the second
drug the first drug should be gradually removed and another is gradually introduced till once
a good dose of the new drug is established. AED should not be withdrawn abruptly. It may
cause rebound of seizure. It should be slow process to avoid the precipitation of withdrawal
seizure. It is generally seen in the barbiturate and benzodiazepines. E.g. Phenytoin 50mg eve-
ry 2 weeks (as a part of a drug change), 50mg every 4 week (total withdrawal). The choice of
the drugs for the different kind of seizure can be listed as below in table 3 as US guidelines.
Table 3: The choice of the drugs for the different kind of seizure as US guidelines.[23]
Type of seizure First line drug Alternative choice
Carbamazepin, Gabapentine,
Lamotrigine, Oxcarbazepine,
Partial seizure( Newly diagnosed) Levetiracetam
Phenobarbitol, Phenytoin, To-
piramate, Valporic acid.
Lamotrigine, Oxcarbazepine,
Partial seizure (refractory Monotherapy)
Topiramate
Generalised seizure absence (newly di-

Lamotigine
agnosed)
Primary generalized (tonic-clonic) Topiramte Lamotrigine, Topiramte
Juvenile myoclonic epilepsy Valporic acid Levetiracete, Topiramate,
Many attempts have been made in the past to obtain anticonvulsant from plant origin and the-
se efforts will continue till a satisfactory treatment becomes available. Besides a number of
allopathic medications available, there is increase in demand for medicinal plants, as these
plants have less side effects.[28]
1.7. Need for the study[37-39]

The use of herbal medicines by physicians in Europe and Asia, exploring their traditional
remedies to find to a suitable cure of these “mind affecting disease” and herbal medicines are
often considered to be gentle and safe alternative to synthetic drugs. Epilepsy is traditionally
treated with anti-seizure medication. But these medications don’t work for everyone, and they
come with risk of side effects. Some herbal have the potential to reduce the seizure.
Flavonoids have the great medical importance and they are found in plant as glycosides.
When they are taken by orally the aglycone part will conjugates by methylation, sulphatation
or glucuronidation. Both the aglycone and conjugates part can pass the blood brain barrier
and bind to the benzodiazepine site of GABAA receptor resulting sedation, anxiolytic or anti
convulsive activity.
β-carotene which is the one of the chief constituents of the leaves of the plant shows the seda-
tive, anticonvulsant, antioxidant, pain relief, antianxiety, antidepressant etc. activity. As for
my knowledge, the anticonvulsant activity on the leaves of the title plant had not done. Tak-
ing these into consideration, there is paucity of antiepileptic activity of the title plant. There-
fore, study was designed for afore said activity.
2. Objectives of the study

The present research work is an attempt to establish the antiepileptic efficacy of leaves ex-
tract of Costus igneus albino mice with the following objectives
1. To collect and authentication of Costus igneus Leaves.
2. To prepare ethanolic extract of Costus igneus Leaves.
3. Test for phytoconstituents of extract
4. Screening for anticonvulsant activity in experimental swiss albino mice by the following
models:
 Electroshock Induced Convulsions
 Pentylenetetrazole Induced Convulsions
 Strychnine Induced Convulsions
3. Review of literature
Review of literature
Distribution and Description
Costus igneus Insulin plant (Costus igneus) is indigenous to Southeast Asia, especially on the
Greater Sunda Islands in Indonesia. It is a relatively new entrant to Kerala and India. The
plant is characterized by large fleshy looking leaves. The undersides of these large, smooth,
dark green leaves have light purple shade. The leaves are spirally arranged around the stem,
forming attractive, arching clumps arising from underground rootstocks. Costus igneus plant
grows very swiftly. Propagation of this plant is by stem cutting. It desires sunshine but it also
grows in vaguely shady areas. Costus igneus does not have a problem with pests and diseas-
es. Outdoor plants might be chewed by caterpillars, and in indoors plants might be preten-
tious by red spider mite.[38] This plant reaches a height of 6 inches and has an indefinite
spread.[39]
Spreading plant reaching about 2 feet tall, with the tallest stems falling over and lying on the
ground. Leaves are simple, alternate, entire, oblong, evergreen, 4-8inches in length with par-
allel venation beautiful, 1-5inch diameter, orange flowers are produced in the warm months,
appearing on cone-like heads at the tips of branches. Fruits are inconspicuous, not showy,
less than 0.5inch, and green colored.[40]
VERNACULAR NAMES
Common name : Fiery costus,Spiral flag
Hindi : Banda, Jarul, keukand
kannada : Kempu honne

Tamil : kostum, Neyccarilcamaram
Telgu : peddavesiga, Yeangesha
Bengali : piasal
Sanskrit : Asana, Bandhukapushpa
Marathi : Honi, pushkarmula
PLANT PROFILE
Kingdom: Plantae
Subkingdom: Viridaeplantae
Domain: Eukaryota
Phyllum: Tracheophyta
Subphyllum: Euphyllophytina
Class: Liliopsida
Subclass: Commelinidae
Order: Zingiberales
Tribe: Coreopsideae
Genus: Costus
Specific epithet: Igneus
Pharmacognosy
Morphological characters
The costus igneus is a rhizomatous shrub and penetrates through the tuberous rhizome. The
rhizome is about 20-40cm grown, cylindrical, soft and fleshy with smooth pale brown
surface. It narrow with several parallel equally thick veins. Top root sub-cylindrical, wider at
the top 28-50cm long,Outer surface light brownish to pale dark brown.
RHIZOMES
Rhizome is circular with smooth and even surface.
LEAF
The leaf is thin with smooth even surfaces, isobilateral and has no difference between the
upper and lower sides. Leaves are simple, alternate, entire, oblong, 4-8 inches long with par-
allel venation, spirally arranged around the stems. Large, fleshy, smooth, and dark green
leaves have a light purple underside.
ROOT
Thin roots have fairly wide superficial sequent periderm and narrow homogenous
parenchymatous cortex.
FLOWERS
. Flowers are orange, 1.5 inches in diameter, on cone-like head at the tips of branches.
PHOTO CHEMISTRY
The following table show the presence of different kind of chemical compound in different
part of Costus igneus plant.
Table 4: Detection of phytoconstituent in different part of plant.

Chemical component Leaves Stem Rhizome
Tannin + + +
Phlotannin + + +
Saponin + + +
Flavonoid + + +
Steroids + + +
Terpenoids + + +
Cardiac glycosides + + +
+Presence
Reported pharmacological activities

Chacko et al, conducted the study of hepatoprotective activity by the alcoholic extract of the
leaves of Costus igneus was used for the experiment. Paracetamol was used in a dose of
300mg/kg orally to induce hepatic damage. The induction of liver damage was confirmed by
elevated serum enzyme levels and histopathological picture showing zonal focal necrosis.
Silymarin was used as reference standard in a dose of 100mg/kg. Administration of Costus
igneus extract prior to Paracetamol administration effectively (P< 0.05) prevented the induc-
tion of damage by Paracetamol, which was confirmed by normal enzyme levels and lack of
necrotic changes in the histopathological studies. The effect produced by 400mg/kg of Costus
igneus was comparable with that produced by the standard drug silymarin. In histopathologi-
cal studies paracetamol treated group showed severe inflammation with focal necrosis. Liver
tissues of rats pretreated with Costus igneus showed almost normal hepatocytes. It was con-
cluded that alcoholic extract of Costus igneus was able to reverse the hepatotoxicity induced
by Paracetamol.[41]
Sardessai et al, conducted the study of antimicrobial activity by the methanolic extract of the
rhizomes of Costus igneus was subjected to the antibacterial studies. The results indicate sig-
nificant activity against both gram-positive (Staphylococcus aureus and Bacillus subtilis) and
gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa and Salmo nella typhim-
rium). The antifungal study revealed no antifungal activity against both Clavi ceps purpurea
and Aspergillus niger. It has been observed that as the concentration of the methanolic extract
of the rhizomes of Costus igneus increases, there was significant inhibition seen in the growth
of the cultures. This is indicated by drastic decline in the absorbance values and confirmed by
plate counts, thus exhibiting strong activity. The present extract ex hibits promising antibacte-
rial and no antifungal acitivity. The above activity has been report ed for the first time from
the methanolic extract rhizomes of Costus igneus.[39]
Danasekaran et al, conducted the study of antiproliferative activity of the plant Costus pictus
was establish to have anti-oxidant and anti-cancer activity .So auxiliary we expanded our
studies on anti-cancer activity of Costus igneus which belongs to the family Costaceae. The
given extract showed potent cytotoxicity against the MCF-7 cell line when compared to the
control group. Danasekaran concluded that the extract showed, it does not impinge on the
normal cell lines. The IC50 value was found to be 750μg/ml. In conclusion the extract has
dose dependent cytotoxicity against the MCF-7 cell line.[39]
Chacko et al, conducted the study of anti hyperlipidemic activity of Costus igneus extract in
Triton X-100 induced hyperlipidemic rats. Administration of tritonX-100 (100mg/kg) to rats
caused an elevation of total cholesterol, total triglycerides, VLDL and LDL and reduction in
HDL levels. Costus igneus was administered at various doses, 100, 200, 400 mg/kg day, (p.o)
to Triton induced hyperlipidemic rats. Atorvastatin was used as reference standard. Treatment
with plant extract was able to significantly (p<0.05) decrease the levels of TC, TG, VLDL
and LDL. Also the extract was found to cause a significant(p<0.05) increase in the HDL lev-
els. The artherogenic index also was decreased in a dose dependent manner. Therefore it can
be concluded that Costus igneus extract was able to effectively suppress Triton induced hy-
perlipidemia in rats, suggesting the potential protective role incoronary heart disease.[42]
Majumdar et al, conducted the study of antibacterial activity, antioxidant activity, anti-
glycation activity of the different parts (leaf, flower, stem and root) of Costus pictus which
exhibited pronounced activity against all tested microorganisms and the activity was quite
comparable with the standard antibiotics screened under similar conditions. The remarkable
antibacterial activity exhibited by the methanolic extracts can be attributed to the synergic
effect of the antimicrobial agents present in it. In conclusion, the data presented here indicate
that the methanolic extracts of flower and stem of Costus pictus possess invitro antioxidant
activity against oxidative protein damage and should be considered as new sources of natural
antioxidants. In this study, Costus pictus extracts showed protective effects against glucose-
induced protein modifications, significantly inhibiting the AGEs formation. Majumdar find-
ings also provide a strong rationale for further investigation to understand the molecular
mechanism of the antiglycation activity of the extracts with both high antioxidant and high
antiglycation activities and explore the possible synergistic antiglycation between them.[43]
Abhirami et al, conducted the study of antifungal activity As antioxidants, flavonoids and
phenolics have been reported to be able to interfere with the biochemical pathways involved
in the generation of reactive oxygen species (ROS), quenching free radicals, chelating transi-
tion metals and rendering them redox inactive in the Fenton reaction. Therefore, the presence
of high amount of flavonoids and other phytochemical compounds along with antifungal ac-
tivity justifies these important medicinal plants of the Islands to be used in traditional and
modern health care practices.[44]
Anyasor et al, conducted the study of anti-inflammatory activity and antioxidant activity of
Costus afer ker Gawl. hexane leaf fraction possesses substantial anti-inflammatory and anti-
oxidant activities against inflammatory diseases especially arthritis. It could be considered as
a choice candidate in pharmaceutical anti-inflammatory drug development.[45]
Palanivel et al, conducted the study of Hypoglycemic activity of Ethanolic extract of Costus
igneus (whole plant) at dose of 250 mg/kg showed significant hypoglycemic activity in allox-
an induced hyperglycemic rats. Thus ethanolic extract of Costus Igneus (Whole plant) is a
multitargeted, cheaper, herbal drug which is a promising candidate for consideration for the
treatment of diabetes mellitus. However, further study is needed to find out the exact mecha-
nism and the phytoconstituents responsible for observed effects.[46]
Alasyam et al, concluded that the Mimosa pudica plant extract possesses anticonvulsant ac-
tivity. However, further studies are necessary to examine underlying mechanisms of anticon-
vulsant activity and to isolate the active compound responsible for these pharmacological ac-
tivities using MES, PTZ and results were found to be significant.[47]
Jay et al, concluded that the methanolic extract of Moringa oleifera leaves possess anticon-
vulsant properties. Further study is required for isolation and identification of active constitu-
ents and to confirm exact mechanism using MES, PTZ and results were found to be signifi-
cant.[48]
Maheshwar Gurunath et al, concluded that methanolic and aqueous extracts of Solanum su-
rattense possessed significant anticonvulsant activity against MES and PTZ seizures than
other extracts when compared to control group.[49]
Plant showing antiepileptic activity

Spondiasmombin. L. (Anacardiaceae): Air dried leaves of the plant had subjected for ex-
traction with aqueous, methanol, and ethanol solvent. Hexobatbital induce sleeping time and
novelty induced rearing (NIR) behaviour in mice and rat were the experiment animal. The
methanolic and ethanolic extract prolonged Hexobarbital inducing sleeping time and reduced
the NIR at dose of 50 and 100 mg/kg. Extract blocked the effect facilitated by the Flimazenil,
which concluded that the effect of Spondiasmombinare not mediated by via muscarinic, α2
adrenergic, and opoid receptor where extract facilitates GABAergic transmission. The extract
block the picrotoxin-induce convulsion due to the presence of phenolic compound in the eth-
anolic and methanolic extract.[29]
Bryonialaciniosa (Cucurbitaceae): The entire plant with the fruits which was shades dried
and made powder into fine and obtained powder was taken into the phytochemical and phar-
macological investigation. It caused the protection of electrically induce convulsion in rat by
39.27%.[30]
Crocus sativusL.(lridaceae): Stigmas of saffron was macerated with ethanol and then anti-
convulsant activity was studied in PTZ induce and MES induce convulsion in the mice. The
alcoholic extract of it decreased the motor activity and prolong the sleep time induced by
Hexobarbital.[31]
Panax ginseng (Araliaceae): Development of the seizure by the injection of the PTZ in al-
ternate days with the 14 injection results the 5 stage of seizure. By o.p. administration of
100mg/kg the Panax ginseng showed the significant protection of seizure. It also caused a the
reduction in the motor activity, eyelid ptosis and bristling fur.[32]
Solanumnigrumlinn (solanaceae): The leaves extract of the Solanumnigrum-

linn(solanaceae) were administered to the 56 rats and 56 chicken before electric shock was
given. The aqueous extract produced a significant protection against electrically induced sei-
zure in rats. Concurrent administration of amphetamine with 30mg/kg of extract produced the
significant protection of 87.5% against electrically induces seizure. Extract at the dose of
40mg/kg of showed the total abolish of the seizure in 3 days. i.p. injection of the extract (10-
60mg/kg) increased the latency of the convulsion induced by PTZ and 30-60 mg/kg extract
showed the complete protection of the seizure and mortality in rates.[33]
Carissa carandasLinn (Apocynaceae): The root of the plant was made into the powder and
extracted with the ethanol by maceration process. This extract was taken into the drying. The
dry product was made soluble with the saline of 80% and 2% of tween. In the electrically in-
duce seizure, the dose of 100,200,400 mg/kg i.p. before 30 min was given to the mice. PTZ
and Picrotoxin were used as then chemical for to induce the seizure in the mice. In the MES
induce seizure model, the dose of 200mg/kg and 400mg/kg of extract protected the mice by
25% and 50%, respectively from seizure whereas in PTZ induced seizure, 200 and 400mg/kg
protect 50% and 62.5%, respectively.[34]
Boerhaaviadiffusa(Nyctaginaceae): The methanolic extract of root of B. diffusa

(1000mg/kg, 1500mg/kg, 2000mg/kg) was given by i.p. prior to 30 min before induction of
the seizure on the mice. In PTZ induced convulsion, the plant extract showed significant pro-
tection, delay in onset of convulsion and reduction in mortality. This may be due to the block-
ing of Ca++ channel by the Liriodendrin.[35]
Clerodendrum infortunatum Linn. (Verbanaceae): The leaves extract was used in the dose
of the 30,50,75, and 100 mg/kg by i.p. routes in the mice. The extract had a saponoin as the
main constitunet, which showed the analgesic action and potentiate the action of the asprine
and pentazocine. It also showed the anticonvulsant action by decreasing the duration of the
seizure and produced a protection in a dose dependent manner against leptazol induce
convulsion.[36]
4. MATERIALS AND METHODS

4.1 Materials
4.1.1. Experimental animals
Species : Albino mice
Sex : Either of sex

Source : Breed and rewired at Bioneeds, Tumkur.
Body weight : 20-30 g
Identification: By cage card and corresponding picric acid colour by body markings.
Number of animals per dose group: 6
Acclimatization: One week in experimental room.
Selection of animals: After acclimatization the animals were subjected to a gross observation
to ensure that the selected mice were in good state of health. mice were randomly selected for
final allotment to the study.
Environmental condition: Air conditioned rooms with optimal air changes per hour, relative
humidity, temperature (20-25ºC) and elimination cycle set to 12 hour light and 12 hour in
dark. The animals were maintained under standard condition in an animal house approved by
the Purpose of Control and Supervision of Experiments on Animals (CPCSEA). The study
protocol was approved by the Institutional Animal Ethics Committee (IAEC), Office of
(IAEC of Mallige College of Pharmacy, Bangalore.
Accommodation: animals are housed in polypropylene cages with stainless steel grill top.
Facilities for food and water bottle and bedding of clean paddy husk.
Diet: “Amrut” brand pelleted feed was provided ad libitum.
Water: UV purified and filtered water was provided ad libitum in polypropylene bottles with
stainless steel sipper tubes.
4.1.2. Apparatus
Table 5: Name of equipment’s.
Sl. No Name of equipment’s Manufacturer
Instruments and chemicals PVT. LTD. Model
1. Convulsiometer
town, Ambala city-134003 (India)
2. Digital balance ACCULAB-Sartorious group
SUPERFIT, Rotatory “vaccum digital bath”
3. Flash evaporator
PMTc-3040
4. 18 gauge oral feeding needle Mallige college laboratory
5. Polypropylene cage Mallige college laboratory
6. Soxhlet Techno scientific, Mumbai
4.1.3. Chemical used

Table 6: Name of chemicals.
SI.NO Names of materials Manufacturer
1. Absolute alcohol Nice – Cochin
2. Phenytion Karnataka fine chem- Bangalore
3. Normal saline Baxter, Aurangabad
4. Pentylenetetrazole Karnataka fine chem- Bangalore
5. Strychinine S D fine Chemicals – Mumbai
6. Diazepam Cadilla Pharmaceutical Limited Gujarat
4.2 Methods
4.2.1. Collection, extraction and preliminary phytochemical studies on Costus igneus
4.2.1.1. Collection and authentication of plant material
The leaves of Costus igneus were collected from Karnataka India. The plant was identified
and authenticated by Prof-Channabasappa, Botanist Sree Siddaganga science of college,
Tumkur, Karnataka, A herbarium specimen was preserved in the college museum for future
reference. The leaves were shade dried at room temperature for 15 days and pulverized.
4.2.1.2. Preparation of extract

The leaves of Costus igneus was dried under shade and then powdered with a mechanical
grinder. The powder was stored in an air tight container for further use.
Extraction procedure
The leaves of Costus igneus will be collected from Karnataka, India. They will be shade
dried. The shade dried powdered leaves (5kg) will be soaked in ethanol (95%) and kept aside
for four days. After four days the ethanol layer is decanted off. The Process will be repeated
for four times. The solvent of total extract is distilled off and concentrate will be evaporated
on a water bath to a syrupy consistency and evaporated to dryness.[41]
4.2.1.3. Preliminary phytochemical studies[41,50]

Ethanolic extract of Costus igneus subjected to qualitative chemical tests for the identification
of their active constituents. Test for the presence of Tannin, Phlotannin, Saponin, Flavonoid,
Steroids, Terpenoids, Cardiacglycosides, were conducted as per the standard procedure. And
the results obtained are presented in table 10.
Chemical tests were carried out on the aqueous extract and on the powdered specimens using
standard procedures to identify the constituents as described by Sofowara (1993) and Har-
borne (1973).
1 Test for tannins

About 0.5g of the powdered samples was boiled in 20ml of water in a test tube and then fil-
tered. A few drops of 0.1% ferric chloride was added and observed for brownish green or a
blue-black colouration.
2 Test for phlobatannins

Deposition of a red precipitate when an aqueous extract of each plant sample was boiled with
1% aqueous hydrochloric acid was taken as evidence forthe presence of phlobatannins.
3 Test for saponin

About 2g of the powdered sample was boiled in 2ml of distilled water in a water bath and fil-
tered. 10ml of the filtrate was mixed with 5ml of distilled water and shaken vigorously for a
stable persistent froth. The frothing was mixed with 3 drops of olive oil and shaken vigorous-
ly then observed for the formation of emulsion.
4 Test for flavonoids

Three methods were used to determine the presence of flavonoids in the plant sample
(Sofowara 1993; Harbrone, 1973). 5ml of dilute ammonia solution were added to a portion of
the aqueous filtrate of each plant extract followed by addition of concen trated Sulphuric ac-
id. A yellow colouration was observed in extract indicated the pres ence of flavonoids. The
yellow colouration disappeared on standing.
In the second method, few drops of 1% aluminium solution were added to a portion of each
fitrate. A yellow colouration was observed indicating thepresence of flavonoids. In the third
method, a portion of the powdered plant sample was in each case heated with 10ml of
ethylacetate over a steam bath for three minutes. The mixture was filtered and 4ml of the fil
trate was shaken with 1ml of dilute ammonia solution. A yellow colouration was observed
indicating a positive test for flavonoids.
5 Test for steroids

Two ml of acetic anhydride was added to 0.5g ethanolic extract of each sample with 2ml sul
phuric acid. The colour changed from violet to blue or green in some samples indicating the
presence of steroids.
6 Test for terpenoids (Salkowski test)

5ml of each extract was mixed in 2ml of chloroform and concentrated sulphuric acid (3ml)
was carefully added to form a layer. A reddish brown colouration of the interface was found
to show positive results for the presence of terpenoid.
7 Test for Cardiac Glyosides (Keller-Killani test)

Five ml of each extract was treated with 2ml of glacial acetic acid containing one drop of fer-
ric chloride solution. It was underlayed with 1ml of conc. sulphuric acid. A brown ring of the
interface indicates a deoxysugar characteristic of cardenolides. A violet ring may appear be-
low the brown ring while in the acetic acid layer, greenish ring may form just gradually
throughout thin layer.
4.2.5. Report of Acute toxicity and dose selection[42]

The maximum lethal dose of Costus igneus same chemical constituents was found to be 2000
mg/kg body weight, hence 1/10th and 1/20th of maximum lethal dose was taken as effective
dose for the ethanolic extract of Costus igneus for antiepileptic activity.
4.2.6. Screening of antiepileptic activity

4.2.6.1. Electroshock Induced Convulsions
Electro-convulsion shock, induce Hind Leg Tonic Extension (HLTE) in 99% of the animal.
The MES test, developed by the Toman and Collaborators more than 60 years ago and proba-
bly the best validated test that predict drugs effective against generalized seizure of the tonic-
clonic type. MES induce tonic extension has been suggested to identify drug that are effective
in preventing seizure spread through neural tissue.[51] Mice weighting 20-30g received an
electrical stimulus of 50 mA for 0.2 sec through auricular electrode to induce maximal sei-
zure of their hind limbs, with tonic extension as the endpoint of the test.
The MES induced convulsion in animal represent grand mal type of epilepsy. This test can
easily conducted and requires a minimal investment in equipment and technical expertise,
and is well standardized.[52]
Experimental design
Electro-convulsive shock, induce Hind Limb Tonic Extension (HLTE) in 99% of the animals.
The electrical stimulus (50 mA for 0.2 sec duration) will be applied through Auricular elec-
trodes using a stimulator apparatus. Animals were divided in 4 groups of 6 animals in each.
Group I were treated with vehicle (0.9% saline). Group II was administered with Phenytoin
(25mg/kg, i.p.). Group III and Group IV were administered with test drug (100mg/kg, p.o.),
(200mg/kg, p.o.) respectively. Electroshock was used to induce epilepsy on 15th day. Imme-
diately after given electroshock, hind limb extension will be observed. Decrease in the dura-
tion of hind limb extension was consisdered as a protective action and determine for each
group.
Table 7: Experimental design for MES model.

SL.no Treatment Duration of Treatment
Vehiclecontrol(0.9%saline)+ Auricular
Group I Daily for 15days+ On 15th day
eletricshock(50mA for 0.2sec)
Phenytoin(25mg/kg, i.p.)+ Auricular
Group II On 15th day+ On 15th day
electricshock(50mA for 0.2sec)
EECI(100mg/kg, p.o.)+ Auricular
Group III Daily for 15days+ On 15th day
electricshock(50mA for 0.2sec)
EECI(200mg/kg,p.o.)+Auricular elec-
Group IV Daily for 15days+ On 15th day
tricshock(150mA for 0.2sec)
i.p= Intraperitonial, p.o=Per oral EECI= Ethanolic extract of Costus igneus
Evaluation
The animals were observed closely for 2 min. Disappearance of the hind leg extensor tonic
convulsion was used as positive criterion. Percent of inhibition of seizures relative to controls
were calculated.
4.2.6.2. Pentylenetetrazole Induced Convulsions

Pentylenetetrazonle (PTZ) has been used widely to induce the convulsion since this chemical
is highly sensitive for comparing different chemical under standard condition.[54] It is consider
as the model for the petit-mal epilepsy. PTZ may elicit seizure by inhibiting ganaergic mech-
anism.[34] The dose of PTZ (80 mg/kg, s.c.) used in this study was determined and found to be
approximate minimal dose(CD100) that induce convulsion in 100% male SWR mice (i.e. con-
trol group). Mice that received PTZ were observed for 30 min.[53]
Experimental design
Swiss albino male mice weighing 20-25g were used. The animals were divided into 4 groups
of 6 each Group I was served as control (0.9% saline). Group II were administered with
standard Diazepam (5mg/kg, i.p). Group III and Group IV were administered with test drug
(100mg/kg p.o) and (200mg/kg, p.o) respectively for 15days. On 15th day PTZ 80mg/kg i.p
was administered to Group I,II,III and IV to induce epilepsy.
Table 8: Experimental design for PTZ model.

SL.no Treatment Duration of treatment
Vehicle control (0.9% saline) + PTZ
Group I Daily for 15 days + PTZ on 15th day
(80mg/kg, i.p).
Diazepam (5mg/kg, i.p.) + PTZ
Group II On 15th day+ PTZ on 15th day
(80mg/kg, i.p).
EECI (100mg/kg, p.o.) + PTZ
Group III Daily for 15days+ PTZ on 15th day
(80mg/kg, i.p).
EECI (200mg/kg, p.o.) + PTZ
Group IV Daily for 15days + PTZ on 15th day
(80mg/kg, i.p).
Evaluation
The time interval between PTZ-injection and occurrence of seizures was measured.
4.2.6.3. Strychnine induce convulsion.

Experimental design
Swiss albino male mice weighing 20-25g were used. The animals were divided into 4 groups
of 6 each Group I was served as control (0.9% saline). Group II was administered with stand-
ard Diazepam (5mg/kg, i.p). Group III and Group IV were administered with test drug
(100mg/kg p.o) and (200mg/kg, p.o) respectively for 15days. On 15th day Strychnine 2mg/kg
i.p was administered to Group I,II,III and IV to induce epilepsy.
Table 9: Experimental design for Strychnine model.

SL.no Treatment Duration of treatment
Vehicle control (0.9% saline) + Daily for 15 days + Strych-
Group I
Strychnine (2mg/kg, i.p). nine on 15th day
Diazepam (5mg/kg, i.p.) + Strychnine On 15th day+ strychnine on
Group II
(2mg/kg, i.p). 15th day
EECI (100mg/kg, p.o.) + Strychnine Daily for 15days+ Strychnine
Group III
(2mg/kg, i.p). on 15th day
EECI (200mg/kg, p.o.) + Strychnine Daily for 15days + Strychnine
Group IV
(2mg/kg, i.p). on 15th day
5.5. Statistical Analysis

All the values were expressed as mean ± SEM. Statistical analysis was performed by one way
analysis of variance (ANOVA) followed by Dunnett’s tests. P values p<0.05, p<0.01,
p<0.001 were considered as statically significant.
6. RESULTS
Table 10: Phytoconstituents present in 95% ethanolic
extract of leaves of Costus igneus.
Serial no. Phytocostitunets Result
1 Tannins +
2 Phlotannins +
3 Saponins +
4 Flavonoids +
5 Cardiac glycosides +
6 Terpenoids +
7 Steriods +
Note:+Present –Absent
Table 11: Effect of ethanolic extract of Costus igneus leaves (EECI) on Maximal
Electroconvulsive Shock (MES) induced seizure in mice.
%Protection %Protection
Group Design of treatment HLTE against against
extensor mortality
I. Vehicle (0.9% Saline) 14.6±0.331 0 50
II. EECI (100mg/kg) 12.5±0.166 16.85 66.66
III. EECI (200mg/kg) 2.31±0.122 85.44 100
IV. Phenytion (25mg/kg) 1.72±0.021 94.76 100
Value are expressed as mean ± SEM (n=6)
Statistical significant test for comparison was done by ANOVA followed by Dunnett’stest
*p<0.05, **p<0.01, ***p<0.001 as compared to control group
D a ta 1
15
c o n tro l
d u r a t io n o f H L T E ( S E C )
EECI I
10 E E C I II
s ta n d a rd
0
1
T r e a tm e n t
Figure 6: Effect of ethanolic extract of Costus igneus leaves on MES induced tonic sei-
zure in mice.
HLTE= Hind Leg Tonic Extension
Each bar represent mean ±SEM (n=6)

Statistical significant test for comparison was done by ANOVA followed by Dunnett’s test
Table 12: Effect of ethanolic extract of Costus igneus leaves (EECI) on PTZ
induced epilepsy in mice.
%Delay in %Protection
Design of Onset of
Group onset of against
treatment seizure
convulsion mortality
Vehicle control
I. 175±6.93 0 0
(0.9% saline)
II. EECI(100mg/kg) 212±6.15 12 33.33
III. EECI(200mg/kg) 215±8.49 22.22 50
IV. Diazepam (5mg/kg) 323±9.9 81.14 83.33
Statistical significant test for comparison was done by ANOVA followed by Dunnett’s test
Figure 7: Effect of ethanolic extract of Costus igneus leaves (EECI) on PTZ induced sei-
zure in mice.
Statistical significant test for comparison was done by ANOVA followed by Dunnett’s
Table 13: Effect of ethanolic extract of Costus igneus leaves (EECI) on Strychnine
induced epilepsy in mice.
Design of Onset of %Delay in onset % Protection
Group
treatment convulson of convulsion of mortality
I. Control(0.9%saline) 294±16.8 0 0
II. EECI(100mg/kg) 333±9.1 22.2 16.66
II. EECI(200mg/kg) 429±12.6 50.06 50
IV. Diazepam (5mg/kg) 457±43.5 75.23 66.66
Figure 8: Effect of ethanolic extract of Costus igneus leaves (EECI) on Strychnine in-
duced convulsion in mice.
5.1 Phytochemical analysis

The Ethanolic (95%) extract of the Costus igneus leaves were subjected to the phytochemical
analysis to identify the presene of the phytoconstituent and result are summerised in (Table
10). The result conform the presence of flavonides, terpenoids, tannins, phlotannins, steriods,
terpinoids,cardiac glycosides, saponines.
5.2 Effect of ethanolic extract of Costus igneus leaves extract(EECI) on MES-induce

epilepsy
Duration of tonic hind leg extension in mice treated with vehicle was 14.6±0.331 seconds.
The EERA at dose of 100mg/kg and 200mg/kg protected the animal from seizure and
significantly reduced the duration of tonic hind lind leg extension to 12.5±0.166 seconds and
2.31±0.122 seconds, respectively. The standard drug phenytion (25mg/kg, i.p.) almost
abolished tonic hind leg extension in the treated animal (i.e. duration of hind led extension
was 1.72±0.0211). Phenytion treatd animal had 94.76% protection against MES induced
seizure where EERA at 200mg/kg and 400mg/kg had 16.85% and 84.44% protection,
respectivey which is shown in table 11 and figure 6.
5.3 Effect of ethanolic extrfact of Costus igneus (EECI) on PTZ induce epilepsy
In the experiment the mice treated with the vehicle (i.e. 0.9% saline) clonic convulsion was
appered at 175±6.93 seconds after PTZ injection via i.p. High dose of EECI (i.e. 200mg)
significantly delayed the onset clonic convulsion for 215±8.49 seconds where as the standard
drug Diazepam (5mg/kg) delayed the onset of clonic convulsion for 323±9.9 seconds.
Diazepam treated animal showed the 81.14% protection against PTZ induced seizure where
as EECI at 100mg/kg, 200mg/kg showed 12% and 22.22% protection respectively by
delaying the onset of seizure.(Table 12 and figure 7)
5.4. Effect of ethanolic extract of Costus igneus leaves (EECI) on strychnine induce
epilepsy
In vehicle treated animal tonic extensior convulsion was appeared at 294±16.8 seconds after
Strychnine injection vai i.p at the dose of 2mg/kg. Similarly at low dose of the extract i.e.
100mg/kg and high dose of extract i.e. 200mg/kg the the tonic extensior convulsion was
appeared at 333±9.1 seconds and 429.2±12.6 seconds, respectively. Standard drug i.e.
Diazepam treated animal showed the tonic extensior convulsion at the 457±43.5 seconds.
Percentage delay in the onset of convulsion was 22.2% for the low dose and 50.66% for high
dose and 75.23% for Diazepam. (table 13 and figure 8)
6. DISCUSSION
Epilepsy is the one of the most common serious neurological conditions. Seizure refres to a
transient alteration of behaviours due to disorder, synchronous and rhythimc firing of
population of brain neurons.[55] Epileptic seizure are common problem of medicinal practice.
The managemant of most epilepsies is fairly satisfactory with the drug availability. However,
a certain percentage of epilepsises do not respond to even polypharmaceutial therapy termed

refractory epilepsy, and the risk of toxicity is very high with such therapy. Also the available
range of drug for use in epilepsy itself indicates that there is no single satisfactory drug to
meet the needs of therapy. Although, advance are being made in the recent years to treat
epilepsy using second-generation drugs. Polypharmay is often advocated to 30% of all
epilepsy patients of refractory or generalized tonic-clonic seizure. Hence necesssary exist for
exploring new drugs, either great anti-seizure activity or less side effect or less interaction
when given combination with other drugs.[56,57]
The most popular and widely used animal seizure model are the traditional MES and PTZ
test. The MES test is the most frequently used as an animal model for the identification of
anticonvulsant activity of drug for the generalized tonic-clonic seizure “grand mal”. PTZ-
induced seizure test is considered as an exprimental model for the “generalizes absence
seizure” and also a valid model for human generalized myoclonic seizure and genaralized
seizure of the petitmal type.[58]
The mechanims of action of AED have been broadly classified into three major category.
Most common form of seizure, namely partial and secondary generalized tonic-clonic
seizures, appears to act by:
1. Reduction or limiting the sustained repetitive firing of neurones, an effect mediated by
promotion or prolonging the inactivated state of voltage-activated Na+ there by reducing
the ability of neurones to fire at high frequencies.
2. Enhancing and faciliting GABA mediated synaptic transmission and inhibition, an effect
mediated either by a pre-synaptic action. In the presence of GABA, the GABAA receptor
is opened, thus allowing an influx of Cl- ion influx in response to GABA or by promoting
GABA release.
Anticonvulsant drugs that are effective against absence seizure, a less common form of
epileptic seizure, act by reducing or limiting the flow of Ca++ channel, thus reducing the
pacemaker Ca++ current that underlies the thalamic rhythm in spikes and wave seen in
generalized absence seizure.[59,60]
MES induce tonic extension can be prevented by AED that inhibit either voltage gated
channels or by blocking glutamergic excitation mediated by the N-methyl-D-aspartame
receptor. AED have the ability to open K+ channel in neuronal cells; can prevent seizure
indce by MES and PTZ.[56]
In our study, it is found that the treatment with EECI significantly reduced in tonic hind leg
extensor stage in MES induced epilepsy in mice. Currently used anticonvulsant drugs (e.g.
Phenytoin, Carbamazepine) effective in therapy of generalized tonic-clonic and partial sei-
zure have been found strong anticonvulsant action in MES test.[58] Since, EERA significantly
inhibited generalized tonic-clonic seizure in MES test; suggest the presence of anticonvulsant
compounds.
Similarly, we found that treatment with EECI in PTZ-induced seizure in mice significanty
reduced the onsest of clonic convulsion. PTZ causes the seizure by inhibiting chloride ion
channels associates with GABAA receptor.[61] Since PTZ has been shown to interact with the
GABA neurotransmission and PTZ-induce seizure can prevent by drugs that enhance
GABAA-receptor-mediated inhibitory neurotransmission such as benzodiazepines and
phenobarbital.[56,61] The effect of the EECI in the PTZ test could therefore suggest
antiepileptic efficacy.
Glycine is a simple aminoacid and an essential intermediates in many metabolic process.

Higher concentration of Glycin is found in spinal cord. Strychinne has the ablility to cause
postsynaptic hyperpolarization by increasing chloride ion concentration. Strychinie produce
the seizure by the excessice exitation of neuronal activity on NMDA receptor.[62-64] So to treat
the convulsion, the test drug should antagonise NMDA-receptor-mediated exitatory
neurotransmission such as Benzodiazepines. In Strychnine induce epilepsy model, we found
that EECI shows the signifcant delay on the onset of tonic extensior seizure convulsion. Thus
EECI may show its activity by inhibiting NMDA receptor.
Antioxidants such as βcarotene, and also saponin have been reported to possess anticonvul-
sant activity.[65,66] The title plant contain the βcarotene as the main compound and also con-
tain saponine.[38,39] The title plant is reported to possess CNS depressant property.[67] It also
contain antioxidant property.[50] Thus, these properties might be playing an important role in
protecting the study animals against seizures.
7. SUMMARY AND CONCLUSION

The present investigation was designed to evaluate the effect of ethanolic extract of Costus
igneus experimentally induced convulsion in mice.
 The leaves of Costus igneus were collected from Karnataka, India.
 Coarse powder of shade dried leaves was extracted with ethanol (95% v/v) using soxhlet
apparatus.
 Three different animal models are used to investigate the effect of title plant:
1. Electroshock induced convulsion
2. PTZ induced convulsion
3. Strychnine induced convulsion
 In MES model, oral administration of EECI at the dose of 100mg/kg and 200mg/kg sig-
nificantly reduce the duration of tonic hind leg extension.
 In PTZ model, oral administration of EECI at the dose of 200mg/kg significantly delayed
the onset of clonic convulsion.
 In strychnine model, oral administration of EECI at the dose of 100mg/kg and 200mg/kg
significantly delayed the onset of tonic extensor convulsion.
In conclusion, ethanolic extract of Costus igneus shows the antiepileptic activity in various
model of epilepsy. It is may be the presence of β-carotene and its derivaties, anti oxident
property and CNS depressent property of plant active constituents antioxidant.
Future Directions: However, further investigation should be carried out to elucidate the exact
mechanism of action.
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Evaluation of Anti-Epileptic Activity of Ethanolic Extract On Swiss Albino Male Mice

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Evaluation of Anti-Epileptic Activity of Ethanolic Extract On Swiss Albino Male Mice

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Divyashree et al. SJIF Impact

EVALUATION OF ANTI-EPILEPTIC ACTIVITY OF ETHANOLIC

*Divyashree L. M. Pharm. and Nagalakshmi N. C.M. Pharm.

Department of Pharmacology, Mallige College of Pharmacy, #71, Silvepura, Bangalore 90.

KEYWORDS: Epilepsy, Costus Igneus, Maximalelectroshock, Pentylenetetrazole,

1.1. Incidence and prevalence of epilepsy

1.2. History of epilepsy[9-11]

1.3. Seizure type and clinical manifestation

1.4. Causes of epilepsy

1.5. Pathophysiology of epilepsy[15-18]

1.6. Treatment of epilepsy[19-21]

1.6.2. General principle of management of epilepsy

 1.6.3. Pharmacological treatment of epilepsy[19-23]

1.6.3.1. Mechanism of action of different AED[24-27]

Figure 2: Representing the different mechanism of action of AED.

Figure 3: GABA receptor with different site for binding of drugs.

Figure 4: Mechanism of action of AED in GABA receptor.

1.6.3.2. Choice of AED

Generalised seizure absence (newly di-

1.7. Need for the study[37-39]

2. Objectives of the study

kannada : Kempu honne

Table 4: Detection of phytoconstituent in different part of plant.

Reported pharmacological activities

Plant showing antiepileptic activity

Solanumnigrumlinn (solanaceae): The leaves extract of the Solanumnigrum-

Boerhaaviadiffusa(Nyctaginaceae): The methanolic extract of root of B. diffusa

4. MATERIALS AND METHODS

Sex : Either of sex

Diet: “Amrut” brand pelleted feed was provided ad libitum.

4.1.3. Chemical used

4.2.1.2. Preparation of extract

4.2.1.3. Preliminary phytochemical studies[41,50]

1 Test for tannins

2 Test for phlobatannins

3 Test for saponin

4 Test for flavonoids

5 Test for steroids

6 Test for terpenoids (Salkowski test)

7 Test for Cardiac Glyosides (Keller-Killani test)

4.2.5. Report of Acute toxicity and dose selection[42]

4.2.6. Screening of antiepileptic activity

Table 7: Experimental design for MES model.

4.2.6.2. Pentylenetetrazole Induced Convulsions

Table 8: Experimental design for PTZ model.

4.2.6.3. Strychnine induce convulsion.

Table 9: Experimental design for Strychnine model.

5.5. Statistical Analysis

Each bar represent mean ±SEM (n=6)

5.1 Phytochemical analysis

5.2 Effect of ethanolic extract of Costus igneus leaves extract(EECI) on MES-induce

a certain percentage of epilepsises do not respond to even polypharmaceutial therapy termed

Glycine is a simple aminoacid and an essential intermediates in many metabolic process.

7. SUMMARY AND CONCLUSION

2005 [cited 2016-1-29]. Available from:

57. Vijayalakshmi A, Ravichandrina V, Anbu J, Malarkodi V, Jayakumari S. Anticonvulsant

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