Evaluation of Antioxidant and Hepatoprotective Effect of Herbal Formulations
Synopsis for registration of M.Pharmacy dissertation.
Submitted to
Rajiv Gandhi University of Health Sciences, Bangalore– 560 041
KARNATAKA
In partial fulfillment
of the requirement for the Degree of
MASTER OF PHARMACY
IN
PHARMACOLOGY
By
Mohan.C.U
I M.PHARM
Under the Guidance of
S. Vimal kumar
Assistant Professor
Department Of Pharmacology
DAYANANDA SAGAR COLLEGE OF PHARMACY
2009
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
BANGALORE - 560 041, KARNATAKA
ANNEXURE - II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. / Name of the Candidateand Address /
Mr. MOHAN C.U
S/o Umesh C.V
“Samrudhi” Sri Drushteshwara swami extension, Chelur, Gubbi taluk, Tumkur -572117
2. / Name of the Institution / Dayananda Sagar College Of Pharmacy
Shavige Malleshwara Hills
Kumaraswamy layout,
Bangalore – 560 078
3. / Course of Study and
Subject / Master of Pharmacy in Pharmacology
4. / Date of Admission / 18th June 2009
5.Title of the Topic: “Antihyperlipidemic effect of Tephrosia calophylla against acute and
Chronic hyperlipidemia in Wistar albino rats.”
6.0
7.0 / Brief resume of the intended work:
6.1-Need of the study:
Hyperlipidemia is metabolic disorder that involves elevation in levels of both cholesterol & triglyceride in plasma. If cholesterol level is raised this is referred as hypercholesterolemia and a raised level of lipoproteins is referred as hyperlipoproteinaemia. The two major clinical squeal of the hyperlipidemia are acute pancreatitis and atherosclerosis. Hyperlipidemia is a major cause of atherosclerosis and atherosclerosis – associated conditions such as coronary heart disease, ischemic cerebrovascular disease and peripheral vascular disease.1Cardiovascular diseases are major cause of mortality all over the world and in Indian subcontinent causing more than 25% of deaths per year. It has been predicted that these diseases will increase rapidly in India to more than half the case of heart diseases in the world within next 15 years. Coronary heart disease & ischemic heart disease have increased in both urban & rural areas. It is mainly due to sedentary life styles and psychosocial stress, high LDL cholesterol, low HDL cholesterol, diabetes, etc.1,2
Lipid & lipoprotein concentrations vary amongst different populations with countries consuming a ‘western diet’ generally having higher calories than those where the regular consumption of fat & cholesterol is low. Many population studies show that the total serum cholesterol level increases with age in both males & females over the age of 20yrs. The treatment of hyperlipidemia patients is by
1. Dietary therapy (less fat, more poly & monounsaturated fat).
2. Exercise (walking, jogging, swimming, cycling).
3. Drug therapy (Antihyperlipidemic agents).
The aims of these therapies are to reduce the serum concentration of elevated atherogenic lipoproteins & triglycerides and to increase the anti-atherogenic HDL levels.
Many drugs are available in market for treating hyperlipidemia, which act by different mechanisms. Synthetics drugs continue to be indispensable against hyperlipidemia. Attempts are pursued to develop drug with minimum side effects. Today’s emphasis of pharmaceutical research & development is on the creation of Therapeutic, Prophylactic and Diagnostic substances with specific functions and minimum side effects. The substances derived from plants have the advantages of being tools for modern medicine, satisfying these conditions. Hence, the advanced countries prefer the natural drugs over synthetic.
Some plant products used as hypolipidemic agent are gugulipid, cholestyramine & colestipol. Further, many literature reviews shows that many plants containing phenolic constituents like Flavonoids are having the hypolipidemic activity. Hence, this study has been designed to investigate the hypolipidemic activity of plants rich in Flavonoids constituents in acute and chronic hyperlipidemia in rat.3
Flavonoids have aroused considerable interest recently because of their potential beneficial effects on human health – they have antiviral, anti-allergic, anti- platelet, and anti-inflammatory and antioxidant activities.4 Many experimental studies in Flavonoids rich plants of genus- Tephrosia have demonstrated antiulcer, hepatoprotective and antihyperlipidemic effects. Hence, in this study I have chosen Tephrosia calophylla plant for hypolipidemic activity6.
Plant description:
Tephrosia PERS. (Leguminosae, Papilionoideae) is a large tropical and sub-tropical genus estimated to contain 300 species7. Tephrosia calophylla BEDD. is a perennial under-shrub found widely in Talakona forest of Andhra Pradesh, South India.8 The genus Tephrosia is known to elaborate a rich variety of Flavonoids and isoflavonoids. Phytochemical investigation of the whole plant of this hitherto university gated species has led to the isolation of a new coumestan,tephcalostan together with two known flavonoids,7-O-methylglabranin and kaempferol-3-O-beta-D-glucopyranoside.9
6.2 – Review of the literature:
Through survey of the literature of the published works- both books and periodicals and internet, yielding the following information
· Ansarullah et.al (2009)4 reported the Hyperlipidemic potential of a polyherbal preparation on Triton WR 1339 (Tyloxapol) induced hyperlipidemia.
· Seru Ganapaty et.al (2009)5 reported the chemical constituents of Tephrosia calophylla Bedd were investigated and a new cytotoxic benzil and coumetan derivatives.
· Pennaka Hari kishore et.al (2003) 9. A new coumestan from Tephrosia calophylla.
· Amit khatri et.al (2009) 12 reported Hepatoprotective activity from aerial parts of Tephrosia purpurea L. and stem bark of Tecomella undulate.
· Bourama naissay et.al (2005) 13 reported Flavonoids from Tephrosia deflexa and T.albifoliolis.
· Damre AS, Gokhale AB, Phadke AS, Kulkarni AR, and Saraf MN (2003) 14 Studies on the immunomodulatary activity of flavonoidal fraction of Tephrosia purpurea.
· Arriaga AM et.al (2009) 15 Antioxidant and larvicidal activities of Tephrosia egregia sandw against Aedes aegypti.
· Pavana P, Manoharan S, Rnju GL, Sethupathy S (2007) 16 reported Antihyperglycemic and antihyperlipidemic effects of Tephrosia purpurea leaf extract in Streptozotocin induced in diabetic rats.
· S.S. Deshpande, G.B. 17 Shah reported Pharmacological activity of Tephrosia purpurea.
· K Soni, P Suresh Kumar, and MN Saraf (2006) 18 reported Antioxidant activity of fraction of Tephrosia purpurea linn.
· K Kavitha, S Manoharan (2006) 19 reported Anticarcinogenic and antilipidperoxidative effects of Tephrosia purpurea (Linn.) Pers. in 7, 12-dimethylbenz (a) anthracene (DMBA) induced hamster buccal pouch carcinoma.
· Cesar C. Andrei .et.al (2000) 20 reported C-prenylflavonoids from roots of Tephrosia tunicate.
· Annalakshmi Chinniah et.al (2009) 21 evaluated the potential of Tephrosia purpurea as anti-Helicobacter pylori agent.
· Palanisamy Prabhakar et.al (1996)22 evaluated a new flavone from Tephrosia hookeriana:
6.3 – Objective of the Study:
The objective of the present investigation is to evaluate the Antihyperlipidemic activity of Tephrosia calophylla against acute and chronic hyperlipidemia.
Plan of work:
1. Collection of plant
2. Extraction
3. Phytochemical Evaluation
4. Hypolipidemic Evaluation
a. Acute Study -
i. Triton WR-1339 induced hyperlipidemia in Rats.
b. Chronic Study -
i. Cholesterol- diet induced hyperlipidemia in Rats.
Materials and Methods:
7.1– Source of Data
Ø Whole work is planned to generate data from laboratory studies i.e. experiments are performed as described in references. Experimental studies in journals and in text books available with college and various institutions.
Data collected from
v Biological abstract
v Chemical abstract
v K R Kirtikar & B D Basu
v Materia Medica
v Medicinal plants of India
v Wealth of India
v Indian journal of pharmacology
v International journal of pharmacology
Websites:
o www.pubmed.com
o www.google scholar.com
o www.academic journals.org
o www.sciencedirect.com
7.2– Method of collection of data:
Plant materials –
Plant material of Tephrosia calophylla will be collected from the perennial under shrub found widely in Talakona forest of Andhra Pradesh. India6. The plant material will be identified taxonomically and authenticated by Dr. S. Madhava Chetty, Asst. Professor Dept. of Botany, Sri Venkateshwara University , Tirupati, Andhra Pradesh. A voucher specimen of the collected sample will be deposited in the departmental herbarium for future reference.
The data and related literatures will be collected from various scientific national and international journal i.e. Indian Journal Pharmacology, Planta Medica, etc.
7.3 -Does the study require any investigations or interventions to be conducted on Patients or other humans or animals? If so, Please describe briefly.
Yes, the study requires investigation on albino rats.
7.4 – Has ethical clearance been obtained from your Institution in case of 7.3?
Yes, Approved by CPSCEA.
7.5– Method:
Preparation of extract :
The air-dried and coarsely powdered whole plant of Tephrosia calophylla (10kg) was successively extracted with polar solvents like n-hexane, Methanol and Dimethylether in a soxhelet apparatus for 8 hr.The crude extract is collected and stored at 20 0C.9
Experimental design:
Evaluation of Antihyperlipidemic activity will be done using following methods:
In the experiment a total number of 24 animals (18 Hyperlipidemic, 6 normal) will be used for each plant extract. The animals will be divided into 4 groups of six each.4
A) Acute Study-
Triton WR-1339 induced hyperlipidemia in Rats:
Group I (G1) : Normal control
Group II (G2) : Hyperlipidemic control
Group III (G3) : Treatment control
Group IV (G4) : Standard control
All animals except normal control group (G1) will be injected with Triton at a dose of 400mg/kg to achieve the Hyperlipidemic animal models.
B) Chronic Study –
Cholesterol- diet induced hyperlipidemia in Rats:
Group I (G1) : Normal control
Group II (G2) : Hyperlipidemic control
Group III (G3) : Treatment control
Group IV (G4) : Standard control
All animals except normal control group (G1) will be fed with cholesterol rich diet to achieve the Hyperlipidemic animal models.
Using these models, the lipid lowering (Hypolipidemic) effect of the extracts can be investigated.
Parameters to be evaluated:
In both acute and chronic hyperlipidemia studies, after the experimental period, all animals will be sacrificed and analyzed for the biochemical parameters in serum & liver tissue and also for histopathological changes which will be carried out by standard method.
1. Biochemical analysis ( Serum & Liver tissue):
a. Total cholesterol
b. Triglycerides
c. Phospholipids
d. HDL cholesterol levels
e. LDL cholesterol levels
f. VLDL cholesterol levels
2. Histopathological analysis:
Diameter & thickness of aorta is measured.
Expected outcome: From these observations, a conclusion may be drawn on the potential of herbs on experimental hyperlipidemia in animal models which can be extrapolated to human beings.
Statistical analysis – The results were expressed as Mean ± SEM. The data was evaluated using one way ANOVA followed by Newman- keuls multiple range test & differences below p<0.05 are considered as significant.
8.0 / List of References:
1. Goodman and Gilman’s; The pharmacological basis of therapeutics; 11th edition 2006 chapter 35, page no 933-966.
2. Aguila-Salinas, C.A., Barrett, H., and Schonfeld, G; Metabolic modes of action of the statins in the hyperlipoproteinaemia; Atherosclerosis, 1998,141:203-207.
3. P.Pavana et.al; Antihyperglycemic and antilipidperoxidative effects of Tephrosia purpurea seed of ethanolic extract in rats; J Environ Biol. 2007 Oct;28(4):833-7.
4. Ansarullah et.al; Hyperlipidemic potential of a polyherbal preparation on Triton WR 1339 (Tyloxapol) induced hyperlipidemia induced hyperlipidemia: A comparison with Lovastatin: International Journal of Green Pharmacy; Apr (2009) 119-124.
5. Seru Ganapaty et.al; chemical constituents of Tephrosia calophylla Bedd were investigated and a new cytotoxic benzil and coumetan derivatives: Phytochemistry; 70 (2009) 95-99.
6. Dr. K. Madhava Chetty, K .Shivji and K. Tulisi Rao: Flowering plants of chittoor district Andhra Pradesh, India, page no. 102.
7. K.R Kirtikar and B.D Basu: Indian medicinal plants vol 3, page no. 724-725.
8. J. S. Gamble: Flora of the Presidency of Madras vol 1, page no. 315-319.
9. Pennaka Hari kishore et.al: A new coumestan from Tephrosia calophylla; Chem. Pharm.Bull. 51(2) 194-196 (2003).
10. Willis J. C., “The Dictionary of Flowering Plants and Ferns” (revised by Shaw, H. K. A.), 8th ed., Cambridge University Press, Cambridge, 1973, p. 1135.
11. Thammanna, Narayana Rao K., Madhava Chetty K., “Angiospermic Wealth of Tirumala,” TTD Press, Tirupati, 1994, p. 49.
12. Amit khatri et.al; Hepatoprotective activity from aerial parts of Tephrosia purpurea L. and stem bark of Tecomella undulate; J. Ethanopharmacol, 122 (2009) 1-5.
13. Bourama naissay et.al; Flavonoids from Tephrosia deflexa and T.albifoliolis; Biochemical systematics and ecology, 33 (2005) 309-312.
14. Damre AS, Gokhale AB, Phadke AS, Kulkarni AR, and Saraf MN: Studies on the immunomodulatary activity of flavonoidal fraction of Tephrosia purpurea; 2003 Apr; Fitoterapia. 2003 Apr; 74(3):257-61.
15. Arriaga AM et.al; Antioxidant and larvicidal activities of Tephrosia egregia sandw against Aedes aegypti; Commun. 2009 Apr; 4(4):529-30.
16. Pavana P, Manoharan S, Rnju GL, Sethupathy S; Antihyperglycemic and antihyperlipidemic effects of Tephrosia purpurea leaf extract in Streptozotocin induced in diabetic rats; 2007 Oct;28(4):833-7.
17. S.S. Deshpande, G.B. Shah; Pharmacological activity of Tephrosia purpurea. AAPS.2008: 50(2).
18. K Soni, P Suresh Kumar, and MN Saraf; Antioxidant activity of fraction of Tephrosia purpurea linn; 2006; 68(4):456-460.
19. K Kavitha, S Manoharan; Anticarcinogenic and antilipidperoxidative effects of Tephrosia purpurea (Linn.) Pers. in 7, 12-dimethylbenz (a) anthracene (DMBA) induced hamster buccal pouch carcinoma: 2006; 38(3):185-189.
20. Cesar C. Andrei .et.al; C-prenylflavonoids from roots of Tephrosia tunicate; Phytochemistry, 55 (2000) 799-804.
21. Annalakshmi Chinniah et.al; The potential of Tephrosia purpurea as anti-Helicobacter pylori agent; Journal of Ethanopharmacology, 124 (2009)642-645.
22. Palanisamy Prabhakar et.al; A new flavone from Tephrosia hookeriana; Phytochemistry, Vol. 43, No. 1, pp.315-316, 1996.
9 / Signature of the candidate / (MOHAN C U)
10 / Remarks of the Guide: / Recommended for research
11 / Name and Designation of:
11.1 Guide:
11.2 Signature: / S VIMAL KUMAR
Asst. Professor
Dept of Pharmacology
Dayananda Sagar College of Pharmacy,
Kumaraswamy layout,
Bangalore – 78.
11.3 Co-Guide:
11.4 Signature / Not applicable
11.5 Head of the Department:
11.6 Signature / GEETHA.K.M
Assoc. Professor &
Head of the Dept.
Dept of Pharmacology
Dayananda Sagar College of Pharmacy,
Kumaraswamy layout,
Bangalore –78.
12 / 12.1 Remarks of the Chairman and Principal / Recommended for research
12.2 Signature / Dr. V. MURUGAN
Professor and principal
Dept of Pharmaceutical Chemistry
Dayananda Sagar College of Pharmacy,
Kumaraswamy layout,
Bangalore –78.