MARRI LAXMAN REDDY INSTITUTE OF PHARMACY

         
 (Approved by AICTE & PCI, New Delhi and Affiliated to JNTUH, Hyderabad)
Dundigal , Quthbullapur Mandal, Hyderabad 500043, R.R. Dist.Telangana.

EVALUATION OF ANTIEPILEPTIC ACTIVITY OF

JATROPHA GOSSYPIIFOLIA LINN BY USING
EXPERIMENTAL ANIMALS

UNDER THE GUIDANCE OF

DR. K. SUNIL KUMAR
Associate Professor ,Department of Pharmacology .

BY
GHATTAMANENI VAISHNAVI (17BU1R0030)
BANDAMEEDHI SWARNA (17BU1R0012)
AKULA AMULYA (17BU1R0001)
BODA VIJAY KUMAR (17BU1R0014)
NEMURI SAI REKHA GOUD (16EG1R0013)
 CONTENTS

 Introduction
 Aim and Objective
 Literature review
 Epilepsy
 JATROPHA GOSSYPIIFOLIA LINN.
 Method and methodology
 Results and discussion
 Conclusion
 Bibliography
 INTRODUCTION
 The word epilepsy is derived from the Greek word meaning “to
seize” or “take hold of”, indicating that the person having a seizure
is “possessed” or at least out of control.
 Epilepsy is a chronic neurological disorder that affects people of all
ages. Around 50 million people worldwide have epilepsy. Nearly
90% of the people with epilepsy are found in developing regions.
 An ideal antiepileptic drug should suppress all seizures without
causing any unwanted effect. Unfortunately, the drugs available in
the modern medicine not only fail to control the seizure activity in
some patients, but quite frequently cause unwanted effects that
range in severity from minimal impairment of the CNS to death
from aplastic anemia or hepatic failure.
 Herbal medicine is currently enjoying a revival in popularity in the
west and in fact it is the primary form of medicine in many parts of
the world.
 J. gossypiifolia is native to the Carribean and tropical America but
is now widespread throughout the tropical world. It has been listed
as a weed in India, Brazil, Jamaica and Trinidad.
 J. gossypiifolia is reported to be beneficial to dyscrasia, anemia,
vertigo and dysphonia. It is an antibiotic, insecticidal and used in
toothache and act as blood purifier.
 In Ghana, decoction of leaves of J. gossypiifolia, Combretum
ghaselensis and the whole part of Ocimun canum are used to
malarial.
 Since after through literature search no data regarding the
antiepileptic activity of the given plant is available. We therefore
have taken this project with the following objectives.
 AIM AND OBJECTIVE
 The main aim is to Evaluate the anti-epileptic activity of Jatropha
gossypiifolia Linn leaves using experimental animal models.
 Objectives:
1. To prepare ethanolic extracts of Jatropha gossypiifolia Linn by
successive Soxhlet extraction technique.
2. To perform phytochemical screening of ethanolic extract of
Jatropha gossypiifolia Linn.
3. To evaluate anti-epileptic activity of Jatropha gossypiifolia
Linn by using following pharmacological experimental animal models.
 Maximum electroshock induced seizure (MES) in mice.
 N-methyl-D-aspartate (NMDA) induced convulsion in mice.
 Pentylenetetrazol (PTZ) induced epileptic seizure in mice
 LITERATURE REVIEW

 Epilepsy is defined as a group of disorders of central nervous

system (CNS) characterized by paroxysmal cerebral dysrhythmia,
manifesting as brief episodes (seizures) of loss or disturbances of
consciousness with or without characteristic body movement
(convulsion), sensory or psychiatric phenomena.
 A “seizure” is a paroxysmal event due to abnormal, excessive,
hypersynchronous discharges from an aggregate of central nervous
system neurons.
 Seizures are controlled in nearly 70% of patients with epilepsy,
mostly through drugs effect on membrane ion channels or on
Gamma amino butyric acidergic (GABA) or glutamatergic
transmission.
 International League Against Epilepsy (ILAE) Classification
of epileptic seizures:

 1. Partial (focal, local) seizures

C. With impairment of consciousness at onset
A. Simple partial seizures (consciousness not
impaired) i. With no other features
ii. With features as in simple partial seizures
i. With motor symptoms
iii. With automatisms
ii. With somatosensory or special sensory
symptoms D. Partial seizures evolving to secondarily
generalized seizures
iii. With autonomic symptoms
iv. Simple partial seizures evolving to generalized
iv. With psychic symptoms seizures.
B. Complex partial seizures (with impairment of v. Complex partial seizures evolving to
consciousness) generalized seizures.
v. Beginning as simple partial seizure and vi. Simple partial seizures evolving to complex
progressing to impairment of consciousness partial seizures to generalized seizures.
vi. With no other features
vii. With features as in simple partial seizures iv.
With automatism
 2. Generalized seizures (convulsive or nonconvulsive):
A. Absence seizures
i. Absence seizures
ii. Atypical absence seizures
B. Myoclonic seizures
C. Clonic seizures
D. Tonic seizures
E. Tonic-clonic seizures
F. Atonic seizures (astatic seizures)
 3. Unclassified epileptic seizures
 ETIOLOGY OF EPILEPSY
 Epilepsy has several causes. Its most likely cause in individual patients relates to age at
onset
A. Idiopathic or constitutional epilepsy
B. Symptomatic epilepsy
i. Pediatric age groups
ii. Metabolic disorders
iii. Trauma
iv. Tumors and other space occupying lesions
v. Cerebrovascular diseases
vi. Multiple sclerosis
vii. Infectious diseases
viii. Degenerative disorders
ix. Genetically determined
 INVESTIGATION, DIAGNOSIS AND
TREATMENT
 Several laboratory studies should routinely be included in the initial diagnostic workup.
Complete blood count
Blood chemistries
Liver and thyroid function tests
 FOR DIAGNOSIS : EEG,CT scan, MRI,FMRI,SPECT,PET.
 FOR TREATMENT :
1. The use of antiepileptic drugs
2. The surgical excision of epileptic foci
3. The removal of causative and precipitating factors
4. The regulation of physical and mental activity
 CLASSIFICATION OF ANTI EPILEPTIC
DRUGS
 First-generation drugs  Second- generation drugs
Phenytoin Lamotrigine
Carbamazepine Topiramate
Valproate Levetiracetam
Ethosuximide Gabapentin
Phenobarbital and Primidone Vigabatrin
Benzodiazepines (e. g. Clonazepam, Felbamate
lorazepam and diazepam) 
Zonisamide
 MECHANISM OF ACTION

      
 JATROPHA  GOSSYPIIFOLIA
 Introduction:  
Family name: Eurphorbiaceae.  
Subfamily : Opuntioideae.  
Genus : Jatropha.
 
 Common name : Bellyache bush, black physicnut or cotton-leaf physicnut
 Vernacular name :
Gujarati name : Ratan Jyoti.
Tamil : Seemayamanakku, Kattamanakku.
Telugu : Nepalam, pepalam
Distribution
This plant is a native of Brazil, naturalized in many parts of India. It grows on
nearly all type of soils within its range.
 Morphology : Leaves are simple, opposite, elliptic or elliptic-lanceolate, smooth,
glossy green, acuminate and wavy margins; flowers are white, sweetly fragrant in
1-8 flowered cymes at the bifurcations of the branches, lobes of corolla
overlapping to right in the bud.
 Phytochemistry: Two triterpenoids have been isolated from leaves of Jatropha
gossypiifolia Linn. Three known flavonoids, vitexin, isovitexin and apigenin
isolated from leaves of Jatropha gossypiifolia Linn. The major phytochemicals of
Jatropha gossypiifolia Linn. are alkaloids, tannins, flavonoids, phenolic
compounds, steroidal sapogenins (saponins)
 Therapeutically leaves are used in Convulsions, stomachache, venereal disease
and as blood purifier.
 There are reported pharmacological activity like Antimicrobial activity, Analgesic
activity, Anti-inflammatory and analgesic activity, Antibacterial activity,
Antimicrobial and Anti-inflammatory activity, Coagulant activity.
 MATERIALS AND METHODOLOGY
 PREPARATION OF PLANT EXTRACT:
Ethanolic extract of plant materials:-
i. Fresh leaves of the plants were collected from the nearby fields of
Gandimaisamma and washed with distilled water so as to remove dust and
foreign particles.
ii. Leaves were then left on the clean surface for shade drying, until well-
dried and grinded to fine powder using a blender and sieved with a 40#
sieve.
iii. After drying the powder, extracted in Soxhlet apparatus for 48 hours with
95% ethanol.
iv. The extract was filtered and dried at room temperature to get a semisolid
mass (yield obtained was: 7.6 % w/w).
v. Stored in air tight container in cool place and used throughout the project
SOXHLET EXTRACTION OF JATROPHA GOSSYPIIFOLIA
WITH 95 % ETHANOL
 PHYTOCHEMICAL SCREENING AND
ACUTE TOXICITY
 The ethanolic extract of Jatropha gossypiifolia (EEJG) was subjected to
phytochemical screening to detect the presence of active constituents such as
alkaloids, tannins, flavonoids, phenolic compounds, steroidal sapogenins
(saponins). The results has been displayed in the table
 The doses of EEJG for pharmacological activity were selected from the literature
i.e 100mg/kg and 200mg/kg.[11]
 EXPERIMENTAL ANIMALS

 Albino mice (20-25g) were used for the anticonvulsant evaluation. They were
procured form Sainath agencies, Musheerabad, Hyderabad (282/99/CPCSEA).
After randomization into various groups and before initiation of experiment, the
rats were acclimatized for a period of 10 days. Animals were housed in
polypropylene cages and maintained under standard environmental conditions
such as temperature (26±2°C), relative humidity (45-55%) and 12 hr. dark/light
cycle. The animals were fed with mice/rodent pellet diet (Golden Mohur Lipton
India Ltd) & water ad libtum. The research was carried out in the experimentation
room of animal husbandry of Marri Laxman Reddy Institute of Pharmacy,
Dundigal, Hyderabad. The study protocol was approved from the Institutional
Animal Ethics Committee (IAEC) before commencement of experiment
(1567/PO/Re/S/11/ CPCSEA)
 EXPERIMENTAL ANIMAL MODELS
GROUPS MES INDUCED SEIZURES NMDA INDUCED PENTYLENETETRAZOLE
IN MICE CONVULSIONS IN INDUCED SEIZURE IN
MICE MICE
GROUP I (normal 1% normal saline 1% normal saline 1% normal saline
control) (1ml/100gm,p.o.) (1ml/100gm,p.o.) (1ml/100gm,p.o.)

GROUP II( pathological Electro convulsiometer NMDA (75mg/kg, s.c) PTZ(70mg/kg, i.p)
control)
GROUP III( positive Phenytoin voil diluted with WFI Memantine diluted with Clonazepam diluted with WFI
control) (25mg/kg, i.p) WFI( 15mg/kg, s.c ) (0.1mg/kg, i.p)
GROUP IV EEJG 1(200mg/kg, p.o in 1% EEJG 1(200mg/kg, p.o in EEJG 1(200mg/kg, p.o in 1%
acacia) 1% acacia) acacia)

GROUP V EEJG 2 (200mg/kg, p.o in 1% EEJG 2(100mg/kg, p.o in EEJG 2(100mg/kg, p.o in 1%
acacia) 1% acacia) acacia)
 RESULTS
 Estimation of % yield of EEJG:
Weight of powder of fresh leaves of Jatropha gossypiifolia Linn. taken = 500gm
Weight of ethanolic extract of Jatropha gossypiifolia Linn. obtained = 38gm
% yield of EEJG obtained = 7.6 % w/w.
S.N0 TEST PRESENCE/ABSENCE
1. Steroids(salkowski test) +
2. Saponin ( foam test) +
3. Flavonoid ( ammonia test) +
4. Triterpenoid (libermann burchard test) +
5. Tannin(fecl3 test) +
6. Cardiac glycosides( keller- killiani test) +
7. Reducing sugars( fehling’s test) +

+ indicates presence
  Anticonvulsant activity of Jatropha gossypiifolia Linn leaves extracts on Maximal
Electroshock induced convulsion in mice.
GROUPS FLEXION EXTENSION CONVULSION STUPOR PROTECTION

PATHOLOGICAL 7.1±0.04 10.60±0.44 17.34±0.43 109.42±0.43 00

CONTROL

PHENYTOIN
SODIUM ( 25mg/kg) 2.16±0.04*** 0.00±0.00*** 7.23±0.44*** 90.96±0.44** 83.33

EEJG – 1 4.33±0.09** 7.35±0.51 15.24±0.50 101.28±0.50 33.33

(100mg/kg)

EEJG – 2 (200mg/kg) 2.55±0.04*** 4.18±0.33*** 13.58±0.47* 105.25±0.47 66.66

EEJG - Ethanolic Extract of Jatropha gossypifolia Linn. All values are expressed as mean ± SEM (n=6),
 Anticonvulsant activity of Jatropha gossypiifolia Linn leaves extracts on NMDA induced
convulsion in mice.

GROUPS DURATION OF TONIC NO OF ANIMALS ANIMALS THAT

EXTENTION (Sec) SHOWS ARE PROTECTED % PROTECTION
TURNING
BEHAVIOUR
AFTER 30 MIN
PATHOLOGICAL 9.50±0.56 6 0 00
CONTROL
MEMANTINE 0.55±0.13*** 0 6 100
(15mg/kg)
EEJG 1 – 7.68±0.27* 2 4 66.66
100mg/kg
EEJG 2– 5.75±0.27** 1 5 83.33
200mg/kg

EEJG - Ethanolic Extract of Jatropha gossypiifolia Linn. All values are expressed as mean ± SEM. (n=6)
  Anticonvulsant activity of Jatropha gossypiifolia Linn leaves extracts on PTZ induced
convulsion in mice.

GROUPS DURATION OF NO OF ANIMALS NO OF ANIMALS

TONIC SHOWS SHOWS % PROTECTION
EXTENTION CONVULSIONS PROTECTION
AFTER 10 MIN
PATHOLOGICAL 12.1±2.57 6 0 00
CONTROL
MEMANTINE 1.2±1.63*** 1 5 83.33
(15mg/kg)
EEJG 1 – 5.2±1.25* 3 3 50.00
100mg/kg
EEJG 2 - 200mg/kg 3.7±1.7** 2 4 66.66

EEJG - Ethanolic Extract of Jatropha gossypiifolia Linn. All values are expressed as mean ±
SEM.(n=6)
 DISCUSSIONS
 Both the doses of the ethanolic extract of Jatropha gossypiifolia Linn. 100
mg/kg and 200 mg/kg protected 33.33% and 66.66% respectively and
standard drug phenytoin sodium 25 mg/kg protected 83.33% mice and
significantly reduced the number of animals convulsing in MES model.
 The ethanol extract of Jatropha gossypiifolia Linn were evaluated at two
doses of 100 and 200 mg/kg body weight, the ethanolic extract of
Jatropha gossypiifolia Linn active agent for anticonvulsant activity with
50% and 66.66% protection of convulsions respectively against
pentylenetetrazol induced seizures with significantly delayed duration of
tonic extensions.
 The ethanol extract of Jatropha gossypifolia Linn were evaluated at two
doses of 100 and 200 mg/kg body weight, the ethanolic extract of
Jatropha gossypiifolia Linn. active agent as NMDA inhibitor for
anticonvulsant activity with 66.66% and 83.33% protection of convulsions
respectively with significantly delayed duration of tonic extension .
 CONCLUSION
 The results obtained in this study indicate that the ethanolic extract
of Jatropha gossypiifolia Linn. have anticonvulsant activities.
 Enhancement of GABAergic neurotransmission and/or calcium ion
channel blockade may be one of the mechanism involved in the
anticonvulsant activity of the extract of Jatropha gossypifolia Linn.
 It is also believed that the presence of various phytoconstituents in
the plant i.e. steroids, saponins, tannins, glycosides, triterpenes,
proteins may be effective for anticonvulsant activities and justify its
use as a traditional folk remedy for central nervous system related
activities.
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