Rajiv Gandhi University of Health Sciences s41
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
KARNATAKA,BANGALORE
ANNEXURE- II
PROFORMA FOR REGISTRATION OF TOPIC FOR DISSERTATION
1. / Name of the CandidateAddress / Dr.Purushotham.K
#554,6th Main Road, Kengeri Satellite town, Bangalore-60,
2. / Name of the Institution / Mysore Medical College & Research Institute,
Mysore-570001
3. / Course of study and subject / M.D. in PHARMACOLOGY
4. / Date of admission to Course / 09 June 2009
5. / Title of the Dissertation / EVALUATION OF ANTICONVULSANT ACTIVITY OF NARDOSTACHYS JATAMANSI IN ALBINO RATS.
6. / Brief resume of intended work:
6.1
6.2 / Need for the Study :
A seizure (from Latin “sacire”, to take possession of) is a paroxysmal event due to abnormal, excessive, hyper synchronous discharges from an aggregate of central nervous system neurons. Epilepsy describes a condition in which a person has recurrent seizures due to a chronic, underlying process. This definition implies that a person with a single seizure, or recurrent seizures, does not necessarily have epilepsy. It refers to a clinical phenomenon rather than a single disease entity, since there are many forms and causes of epilepsy. 1
Using the definition of epilepsy as two or more unprovoked seizures, the incidence of epilepsy is ~ 0.3-0.5 percent throughout the world and the prevalence has been estimated at 5-10 persons per 1000. 1 Epilepsy is a major public health issue in many nations and the frequency of epilepsy and seizures increases in later life. Despite the massive scale of the problem and much research, epilepsy remains poorly understood. Despite more than 20 approved drugs and several non-pharmacological options, up to 30% of patients are still refractory to treatment. Despite over a century of pharmacotherapy and neuroscience research, rational design of anti-epileptic drugs is only now starting to yield results, because of the heterogeneity of the disease and our limited understanding of it. Discovery and development of anti-epileptic drugs has been especially difficult, because of the regulatory issues of satisfactorily proving safety and efficacy, ethical constraints on placebo-controlled trial designs, the fact that seizures are typically widely spaced in time, and the fact that the person undergoing the seizure is typically in no state to remember, let alone assess, what happened. Hence there is ongoing search for newer therapeutic products for the treatment of neurological disorders.
Medicinal plant research worldwide is progressing constantly, demonstrating the pharmacological effects in neurological disorders. A number of single and compound drug formulations of plant origin are mentioned in Ayurveda, for the treatment of neurological disorders like epilepsy.
Review of Literature: Approximately 1 percent of the worlds population has epilepsy, the second most common cause of neurological disorder after stroke. For long time it was assumed that a single drug could be developed for the treatment of epilepsy. However, the cause of epilepsy is diverse.
Sodium Valproate is used as standard drug in this study. It is active against both Pentylenetetrazole and Maximal Electroshock seizures.2It causes a significant increase in Gamma Amino Butyric Acid content of brain. It enhances action of Gamma Amino Butyric Acid by postsynaptic action. It also inhibits sodium channels.3 In ayurveda, herbal drugs like Bacopa monniera (Brahmi), convolvufus pluricaulis (Shankhpushpi) and Nardostachys jatamansi (Jatamansi) are used as medicine in the treatment of various mental disorders. They act as tranquilizers and also used in the treatment of hypertension.4
Spikenard (Nardostachys jatamansi; also called nard, nardin, and muskroot) is a flowering plant of the Valerian family that grows in the Himalayas of China, India and Nepal. The plant grows to about 1m in height and has pink, bell-shaped flowers. Spikenard rhizomes (underground stems) can be crushed and distilled into an intensely aromatic amber-coloured essential oil, which is very thick in consistency.
The rhizomes and roots significantly affect amylase activity. The root is also used in perfumery. The rhizomes of Jatamansi are used as bitter tonic , stimulant and antispasmodic, as well as to treat epilepsy, chorea and palpitation.5
Common uses - as antipyretic, burning sensation, erysipelas, leprosy, skin diseases, throat inflammation, ulcers, intestinal colic, as an antispasmodic, diuretic, carminative, laxative, and in hysteria and cholera. It is a component of the Ayurvedic anti-epileptic drug “Ayush-56 ”.6
It was pharmacologically investigated in animal experiments of sedative, tranquilizing and antihypertensive properties. In some experiments, typical for tranquilizers, certain activities could be demonstrated such as the prolongation of barbiturate hypnosis, the impairment of rotarod performance, an anticonvulsive activity on electric shock and potentiation of the body-temperature lowering activity of Reserpine.7
6.3
7.
7.1
7.2.
7.3
7.4
8 / Objectives of the Study:
1. To evaluate the anticonvulsant activity of Nardostachys jatamansi in albino rats.
2. To compare the anticonvulsant activity of Nardostachys jatamansi and Sodium Valproate in albino rats.
Materials and Methods :
Source of data :
Animal experiments in albino rats in the Department of Pharmacology,Mysore Medical College and nd Research Institute.
Albino rats weighing around 150 g – 200 g of either sex will be randomly selected from
central animal facility, Mysore Medical College and Research Institute, Mysore.
Method of collection of data : Animal experiments in the Department of Pharmacology.
INCLUSION CRITERIA
1. Albino rats weighing 150 to 200 g of either sex.
2. Animals acclimatized to the conditions of experiments 24 hours before experiment.
EXCLUSION CRITERIA
1. Pregnant animals.
2. Diseased animals.
CHEMICALS USED
1. Nardostachys jatamansi root extracts .
2. Sodium Valproate .
3. Appropriate solvent.
Ø Nardostachys jatamansi standardized ethanolic root extract is used for experiments. 8
Ø Nardostachys jatamansi extract and Sodium Valproate are dissolved in appropriate solvent.
Equipments
q Electro-convulsiometer with accessories.
q Mililitre syringes (having 100 divisions).
q Measuring jars and glass beakers.
q Weighing balance.
q Animal (albino rat) weighing balance.
q Animal cages.
Albino rats weighing around 150 g – 200 g of either sex will be randomly selected from central animal facility, Mysore Medical College and Research Institute, Mysore. Animals will be provided free access to water and animal food, and are maintained under standard laboratory conditions with a natural light and dark cycle, under room temperature. The animals will be acclimatized for 24 hours before the start of experimentation.
Sample size: Animals are divided into 10 groups of 6 albino rats each.
For Maximal Electroshock Seizure model :
Group 1 - Control group treated with appropriate solvent .
Group 2 - Standard group treated with Sodium Valproate.
Group 3 - Test group treated with dose 1 of Nardostachys jatamansi extract.
Group 4 - Test group treated with dose 2 of Nardostachys jatamansi extract.
Group 5 - Test group treated with dose 3 of Nardostachys jatamansi extract.
For Pentylenetetrazole Induced Convulsion model :
Group 6 - Control group treated with appropriate solvent .
Group 7 - Standard group treated with Sodium Valproate.
Group 8 - Test group treated with dose 1 of Nardostachys jatamansi extract.
Group 9 - Test group treated with dose 2 of Nardostachys jatamansi extract.
Group 10 - Test group treated with dose 3 of Nardostachys jatamansi extract.
Laboratory models for testing anticonvulsive activity :
1.Maximal electroshock induced seizures:-The maximal electroshock model is the best studied and most useful animal model of seizures. In this test, brain receives a maximal or supramaximal electrical stimulus which results in generalized convulsive seizures. The profile of maximal seizures includes tonic flexion of all fore limbs followed by tonic extension of hind limbs followed by terminal generalized tonic-clonic convulsions. Using a electroconvulsiometer electrical stimulus of 150 mA in rats will be applied via ear clip electrodes for a period of 0.2 seconds. 24 hours before testing of anticonvulsants, the animals are pre-screened for their ability to develop full tonic extension in the maximal electroshock test. Abolition of the hind limb extensor component after treatment with the test drug is considered as the end point. The tonic component is judged abolished if the hind limb extension does not exceed a 90 degrees angle with the plane of the body. The maximal electroshock test identifies drugs such as phenytoin, carbamzepine and lamotrigine, which are active against generalized tonic-clonic seizures and complex partial seizures.
GROUPS / LP (sec) (mean) / TEHL (sec) (mean) / RECOVERY PERIOD(sec) (mean)
Group 1
Group 2
Group 3
Group 4
Group 5
LP – Latency period. TEHL – Tonic extension of hind limbs.
2.Pentylenetetrazole (PTZ) Induced Convulsion: This assay is one of the most commonly employed method for anticonvulsant screening. Pentylenetetrazole is a central nervous system stimulant. The convulsive effect is analogous to petit mal type of convulsion in man. The test compound and the reference drug is injected intraperitonially, to the groups of albino rats. Another group of same number of rats serves as control. Thirty minutes after intraperitonial injection, 60-90 mg/kg Pentylenetetrazole is injected intraperitonially. Each animal is placed into an individual plastic cage for observation lasting 30 minutes. Pentylenetetrazole initially produces myoclonic jerks, which becomes sustained and lead to generalized tonic-clonic seizures. The end points include:
1.The first generalized clonic seizure with loss of righting reflex,
2.The first episode of continuous generalized clonic seizure with loss of righting reflex for atleast 5 seconds,
3.Onset of myoclonic jerks,
4.Straub’s tail. Complete abolition of the chosen end point can be considered as 100 percent protection and indicates anti- convlsant activity of the test compound.
The number of protected animals in the treated groups is calculated as percentage of affected animals in the control group. The time interval between Pentylenetetrazol injection and occurrence of seizures can be measured. The delay of onset is calculated in comparison with the control group.
GROUPS / ONSET OF MYOCLONIC SPASM (sec) (mean) / ONSET OF CLONIC CONVULSIONS (sec)(mean)
Group 6
Group 7
Group 8
Group 9
Group 10
Statistical Analysis:
· The data will be analysed in SPSS 16 version for Windows.
· Student “t” test will be used.
· The effects of different drugs under study will be presented by calculating descriptive statistics like Mean and S.D of the outcome parameters.
· Analysis of Variance (ANOVA) will be applied to compare the effects of the drugs under study.
· Tests of significance will be carried out at 5% level (p < 0.05 ).
Does study requires any investigation or intervention to be conducted on humans or animals?
Yes.
Has ethical clearance been obtained from your institution?
Yes.
List of References
1. Daniel H. Lowenstein. Seizures and epilepsy. Harrisons Princples of Internal Medicine,2009; 17th edition, Mc Graw Hill : pp 2498.
2. Roger J and Brain S.Meldrum. Antiseizures Drugs. Basic and Clinical Pharmacology, 2009; 11th edition, Tata-McGrawHill : pp 400.
3. H.P. Rang , M.M. Dale, J.M. Ritter , R.J. Flower. Antiepileptic drugs. B Rang and Dales Pharmacology ,2007; 6 th edition, ChurchHill-Elsevier : pp 575 – 587.
4. Indurwade NH, Biyani KR. Evaluation of comparative and combined depressive effect of Brahmi, Shankhpushpi and Jatamansi in mice, Indian J Med Sci 2000; 54: pp339-41.
5. Rajkumar Subhashini, Arunachalam Gnanapragasam, Subramananian Senthilkumar; Protective Efficacy of Nardostachys Jatamansi on Mitochondrial Respiration and Lysosomal Hydrolases during Doxorubicin Induced Myocardial Injury in rats, Journal of Health Science,2007 ; vol 53(1) : pp 67.
6. Joshi, Hanumanthachar , Parle, Milind ; Journal of medicinal food (J Med Food), published in United States, 2006, vol 9 (1) : pp 113-8.
7. Rücker G, Tautges J, Sieck A, Wenzl H, Graf E. Isolation and pharmacodynamic activity of the sesquiterpene valeranone from Nardostachys jatamansi DC, Arzneimittelforschung. 1978;vol 28 (1): pp 7-13.
8. Rao.V.S,Rao .A,Karanth .K.S . Anticonvulsant and Neurotoxicity profile of Nardostachys Jatamansi . J Ethnopharmacol,2005 dec 1;102(3) : pp 351-355.
9. / Signature of the Candidate / ( DR.PURUSHOTHAM.K )
10. / Remarks of the Guide
11. / Name & Designation of
11.1 Guide / DR. BASAVANNA .P.L.
PROFESSOR ,
DEPARTMENT OF PHARMACOLOGY,
MMC & RI , MYSORE .
11.2 Signature
11.3 Co-Guide (If any) / NIL
11.4 Signature / NIL
11.5 Head of Department / DR.PARASHIVAMURTHY,
PROFESSOR AND HEAD OF DEPARTMENT,
DEPARTMENT OF PHARMACOLOGY,
MMC & RI , MYSORE .
11.6 Signature
12. / 12.1 Remarks of the
Dean Director
12.2 Signature