ANTICONVULSANT STUDIES OF THE METHANOL EXTRACT AND FRACTIONS OF CASSIA SIAMEA LAM. (FABACEAE) IN MICE AND CHICKS

TABLE OF CONTENT
Title page
Abstract
Table of Contents

CHAPTER ONE
1.0       INTRODUCTION
1.1       Statement of research problem
1.2       Justification
1.3       Theoretical framework
1.4       Aims and objectives
1.5       Research Hypothesis

CHAPTER TWO
2.0       LITERATURE REVIEW
2.1       Etiology and risk factors for developing Epilepsy
2.2       Classification of seizures
2.2.1    Partial seizures
2.2.2    Generalized seizures
2.2.3    Unclassified epileptic seizures
2.3       Mechanism of Epileptogenesis
2.3.1    Basic Mechanisms of Focal Seizure Initiation and Propagation
2.3.2    Gamma aminobutyric acid (GABA) - receptor Inhibition
2.3.3    Glutamate Receptors Activation
2.3.4    Extra-neuronal (extrinsic) factors
2.4       Diagnosis of Epilepsy
2.4.1    Electroencephalography
2.4.2    Neuro-imaging Computerized Tomography
2.4.3    Plain Radiology
2.4.4    Single Photon Emission Computerized Tomography (SPECT)
2.4.5.   Magnetic Resonance Imaging (MRI)
2.5       Classification of Antiepileptic drugs on the basis of their mechanism(s) of action
2.5.1    Voltage-Gated Sodium Channels blockers
2.5.2    Voltage-gated Calcium channels blockers
2.5.3    GABA ergic agents
2.5.4    Glutamate blockers
2.5.5    Carbonic anhydrase inhibitors
2.6       Limitations associated with the use of existing antiepileptic drugs
2.7.1    Medicinal plants and Epilepsy
2.7       Plant description: Cassia siamea
2.6.1    Botanical description
2.6.2    Taxonomic classification
2.6.3    Ethnomedical uses of Cassia siamea plant in

CHAPTER THREE
3.0       MATERIALS AND METHODS
3.1       MATERIALS
3.1.1    Plant Material collection and Authentication
3.2       Experimental animals
3.3       Drug and chemicals
3.4       Equipment
3.5       Fractionation of Cassia siamea crude extract
3.6       Preliminary phytochemical screening
3.3       Pharmacological Studies
3.3.1    Acute toxicity Studies (LD50)
3.3.2    Anticonvulsant studies
3.3.2.1 Maximal electroshock-induced seizure test in chicks
3.3.2.2 Pentylenetetrazole –induced convulsion in mice
3.3.2.3 Picrotoxin-induced seizure in mice
3.3.2.4 Effect of naloxone on the anticonvulsant activity of the Ethyl acetate fraction
3.4       Behavioral study on the Ethylacetate fraction of Cassia siamea plant extract
3.4.1    Diazepam-induced Sleep test in Mice
3.6       Statistical Analysis

CHAPTER FOUR
4.0       RESULTS
4.1.      The Percentage yield of the Plant Extract and Fractions
4.1.1    Fractionation of the Crude Methanol Whole Plant Extract
4.2       Phytochemical constituents of the Whole Plant Extract of Cassia siamea
4.3       LD50 Value Determination in Mice and Day-Old Chicks
4.4       Anticonvulsant Studies
4.4.1    Maximal Electroshock-induced Seizures in Chicks
4.4.2    Pentylenetetrazole- induced seizures in mice
4.4.3    Effect of the Ethyl acetate fraction of the Methanol Extract of Cassia siamea on Picrotoxin-induced Seizure in Mice
4.4.4    Effect of Naloxone on anticonvulsant activity of the Ethyl acetate portion of Cassia siamea on Pentylenetetrazole (S.cPTZ) –induced convulsion in mice
4.4.5    Effect of the Ethyl acetate fraction of Cassia siamea on Diazepam induced Sleep

CHAPTER FIVE
5.0       DISCUSSION


CHAPTER SIX
6.0       SUMMARY, CONCLUSION AND RECOMMENDATIONS
REFERENCES

ABSTRACT
Cassia siamea is a shrub belonging to the Fabaceae family, native of Southeast Asia. The plant is commonly used in traditional medicine to treat hypertension, malaria and diabetes. Due to the easy cultivation of the plant, its widespread and also remarkable biological activities, Cassia siamea has become a worldwide medicine. The study was conducted to assess the anticonvulsant potential of the methanol extract of Cassia siamea (CSME) and its fractions (EAF, NBF and RAF) using Pentylenetetrazole (PTZ) induced seizures in mice and Maximal electroshock (MES) induced seizures in chicks. Acute toxicity was carried out on CSME and fractions. The possible mechanism(s) involved in anticonvulsant action were determined using Picrotoxin and naloxone. The preliminary phytochemical screening of the methanol extract revealed the presence of alkaloids, flavonoids, polyphenols, saponins, steroids, terpenoids and tannins. The LD50 of CSME, its EAF and NBF was found to be greater than 5000mg/kg intraperitoneally (i.p) in chicks and mice, suggesting a non toxic profile of the extractst, while the LD50 of the RAF (i.p) was found to be 1095mg/kg in mice and 3807 mg/kg in chicks implying the RAF is mildly toxic via the intraperitoneal route. The CSME and its EAF were found to have varied anticonvulsant activities; the EAF at 250mg/kg dose protected 40% of mice against hind limb tonic extension induced by maximal electroshock and in convulsed chicks a significant (P < 0.05) decrease in mean recovery time was noted. The ethyl acetate fraction at 250mg/kg and 500mg/kg doses produced a significant (P < 0.05) delay in mean onset of seizures and offered mice a 2/6 and 5/6 quantal protection against mortality, valproic acid the standard anticonvulsant drug used produced a 100% protection against seizures. The EAF did not protect mice against pirotoxin induced seizures indicating lack of activity on chloride ion channels of the GABAA receptor complex. Naloxone did not reverse the anticonvulsant activity of the EAF against Pentylenetetrazole-induced seizures, suggesting lack ogf involvement of GABAA-BDZ receptors and opioid receptors in the anticonvulsant effect of EAF. The CSME and EAF at 100mg/kg dose significantly (P < 0.05) prolonged the total duration of Diazepam induced sleeping time in mice without affecting the mean onset of sleep, indicating sedative action of the extract. The results suggests the presence of bioactive component(s) that posess anticonvulsant and sedative activities. The data may provide pharmacological basis for the use of the plant alone or in combination with other plant(s) in the management of febrile convulsions and insomnia in West African countries including Nigeria.

CHAPTER ONE
1.0  INTRODUCTION
1.1  Background information
Epilepsy is one of the most common afflictions of human beings (Muralidharanet al., 2009). It affects approximately 50 million people worldwide and accounts for about 1% of the global burden of disease (Reynolds, 2002). Epilepsy is coined from the Greek word

epilembanein” which means "to seize”. It is a chronic neurological disorder affecting both sexes (Blume et al., 2001). It is the second most common chronic neurological condition seen by neurologists worldwide (Sridharan, 2002). It is characterized predominantly by recurrent and unpredictable interruptions of normal brain function, called epileptic seizures which arise due to sudden, excessive and rapid discharge of cerebral neurons in the grey

matter of the brain. It is a diverse family of disorders, having in common an abnormally increased predisposition to seizures reflecting underlying brain dysfunction that may result from different causes (Fischer et al., 2005).

Among brain disorders, epilepsy stands out not only because of its high prevalence and incidence rates, but in particular because of the myths and beliefs attached to the condition in various cultures and the resulting impacts on the individual, the family, and the community as a whole (Jamison et al., 2006). Epilepsy remains among the most stigmatized brain disorders. The associated stigma is more obvious in developing countries because of illiteracy and misinformation regarding the actual nature of the condition....

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