“ANTI-INFLAMMATORY ANDANTI-ARTHRITIC ACTIVITY OF AMORPHOPHALLUS PAEONIIFOLIUS”

M. Pharm Dissertation Protocol Submitted to

Rajiv Gandhi University of Health Sciences, Karnataka

Bangalore– 560041

By

Mr. Bharat Bhushan PandeyB.Pharm

Under the Guidance of

Dr. Preeti KulkarniM.Pharm, Ph.D

Department of Pharmacology

S.E.T’s COLLEGE OF PHARMACY

S. R. Nagar, Dharwad–580002

2011-2012

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

BANGALORE, KARNATAKA

ANNEXURE –II

PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION

1. / NAME OF THE CANDIDATE AND ADDRESS / BHARAT BHUSHAN PANDEY DEPT. OF PHARMACOLOGY, SET’s COLLEGE OF PHARMACY, S.R.NAGAR, DHARWAD-580002
2. / NAME OF THE INSTITUTION / SET’s COLLEGE OF PHARMACY, S. R. NAGAR, DHARWAD-580002
3. / COURSE OF STUDY AND SUBJECT / MASTER OF PHARMACY IN PHARMACOLOGY
4. / DATE OF ADMISSION TO THE COURSE / 11/07/2011
5. / TITLE OF THE TOPIC:
“ANTI-INFLAMMATORY AND ANTI-ARTHRITIC ACTIVITY OF AMORPHOPHALLUS PAEONIIFOLIUS”
6.0
7.0
8.0 / BRIEF RESUME OF THE INTENDED WORK
6.1Need for the study :
Inflammation is a normal response to any noxious stimulus that threatens the host and may vary from localized response to a generalized one1. The inflammatory process protects our body from diseases by releasing cells and mediators that combat foreign substances and prevent infection2. Inflammation is a complex process involving various mediators, such as prostaglandins, leukotrienes and platelet activating factors. Inflammation is caused by release of chemicals from tissues and migrating cells3. In acute stages, the inflammatory process is vital to the healing process4.
Arthritis is inflammation of one or more joints. A joint is the area where two bones meet. There are over 100 different types of arthritis. Basicallyarthritis involves the breakdown of cartilage(normally protects a joint, allowing it to move smoothly), that also absorbs shock when pressure is placed on the joint, as during walking. Without the normal amount of cartilage, the bones rub together, causing pain, swelling (inflammation), and stiffness and limited movement.Arthritis may occur in men or women. The need of treatment is to reduce pain, improve function, and prevent further joint damage5.
Nowadays, animal models of rheumatoid arthritis are usedextensively in research on pathogenesis of inflammatory arthritis and in the pharmaceutical industry in the testing of potential anti-arthritic agents6.
The history of anti-arthritic therapyfrom non-steroidal anti-inflammatory agents to biological response modifiers is placed in context with the evolution ofanimal models of arthritis7.Amorphophallus Paeoniifolius(Family-Araceae) is a herbaceous plant with underground hemispherical depressed dark brown corm. The corms of plant are acrid,astringent,thermogenic,irritant,anodyne,anti-inflammatory,anti-haemorrhoidal, haemostatic, expectorant, carminative, digestive, appetizer and stomachic8. The plant has been reported to have anti-inflammatory9, antidiarrhoeal10, gastroprotective11, anthelmintic12 activity .Whether these claims are valid or not should be probed scientifically to establish the rational use.
The present study is designed to evaluate the effect of above tuber on chronic inflammatory disease, i.e., arthritis.
6.2 Review of Literature :
Amorphophallus Paeoniifolius (Family: Araceae) is cultivated in Karnataka, West Bengal, Maharashtra and in plains of North India. Leaf solitary, large, Spathe bell-shaped, greenish pink inside, purplish outside. Flower-unisexual, berried-clustered, obvoid-red, Fruits-ovoid 2-3 seeded with red colour8.Corm of the plant contains lupeol, β-sitosterol, stigmasterol, betulinicacid, quercetine, triacontane, β-sitosterol palmitate, galactose, glucose, rhamnose and sucrose13.Roots are used in opthalmia, boils and as emmenogogue. Fermented juice of petiole are used to cure diarrhea. Seeds are used in rheumatism8.
6.3Objectives of Study :
1. To study the anti-inflammatory activity of Amorphophallus paeoniifolius on experimental animals by using the following models
  • Formalin-induced paw oedema model
  • Cotton pellet granuloma model
2. To carry out anti-arthritic activity of Amorphophallus paeoniifolius on experimental animals by using
  • Complete Freund’s adjuvant induced arthritic model
Materials and methods
7.1Source of Data :
The source of data are
  • Laboratory experiments on animals
  • Literature survey
  • Books and journals
  • Internet
7.2Method of collection of data:
Materials :
  • Collection, identification and authentification of the plant by a Botanist.
  • Chemicals and Drugs: Petroleum Ether (60-80ºC), Ethanol 90%, Diclofenac sodium
Extraction :
  • Shade dried corm of Amorphophallus paeoniifolius will be extracted successively with PetroleumEther (60-80ºc) and 90% ethanol in a soxhlet’s apparatus.
Animals :
Albino Wistar rats weighing 150-200gm will be used. The animals will be maintained under controlled condition of room temperature and 12h light-dark cycles. The animals are randomized into different groups and housed in sanitized polypropylene cages containing sterile paddy husk. They will have free access to standard pellets as basal diet and water ad libitum.
Pharmacological studies :
The animals are grouped as follows for carrying out above mentioned activities.
Group / Treatment
Group 1 (normal) / Vehicle(Distilled water)
Group 2 (positive control) / Inducing agents (Formalin,Cotton pellet and Complete Freund’s Adjuvant)
Group 3 (petroleum ether extract lower dose) / Test drug + inducing agent
Group 4 (petroleum ether extract higher dose) / Test drug + inducing agent
Group 5 (ethanolic extract lower dose) / Test drug + inducing agent
Group 6 (ethanolic extract higher dose) / Test drug + inducing agent
Group 7 (Standard) / Standard drug + inducing agent
  1. Anti-inflammatory activity
a. Formalin-induced oedema in rat hind paw14:
Albino wistar rats of either sex will be divided into seven groups (5 in each) as given in above table and treated with drugs 60 minutes before formalin injection (20µl of 2.5%w/v in distilled water). Formalin will be injected into sub plantar tissue of left hind paw of each rat. Swelling of formalin injected paw volume will be measured at different time interval such as 0 hr, 30 min, 1, 2, 3 and 4 hr using Plethysmometer. The right hind paw will be injected with 0.1 ml of vehicle. Diclofenac sodium serves as reference standard.
b. Cotton pellet granuloma in rats15:
In this method, the autoclaved cotton pellets 50±1 mg will be implanted subcutaneously by incision on the back of rats under ether anaesthesia. Drugs will be administered orally for 7 days. Animals will be sacrificed on day 7 and the granuloma will be dissected out, dried in an oven at 600C and weighed to determine thepercent inhibition of granuloma andcompared with standard (Diclofenac sodium).
2. Anti-arthritic activity
  • Complete Freund’s Adjuvant (CFA) induced arthritis in rats16:
Male Wistar rats weighing 150-200gm will be selected for the experiment. They will be divided into different groups as per the above table. On day ‘0’, the basal paw volume of left hind paw of each animal will be measured using mercury plethysmometer. On the 1st day all the animals except normal group will be anaesthetized, they will be injected into the sub plantar region of left hind paw with 0.1 ml of CFA (Sigma Aldrich, USA) containing 0.1 mg of heat killed Mycobacteriumtuberculosis cells in liquid paraffin and will be allowed to recover to serve as control. Treatment with Dexamethasone and extracts will be started on the same day i.e. 1st day and continued for 21 days. Normal group will receive saline. Paw volume will be measured on 4th, 8th, 14th and 21st day of study period.
Statistical analysis:
The data obtained will be expressed in MEAN ± SEM and analysed by one way ANOVA followed by Tukey’s test.
7.3 DOES THE STUDY REQUIRE ANY INVESTIGATION OR INTERVENTIONS TO BE CONDUCTED ON PATIENTS OR OTHER HUMANS /ANIMALS? IF SO PLEASE DESCRIBE BRIEFLY.
Yes, the above study requires investigation to be done on the rats and mice for the determination of anti-inflammatory and anti-arthriticactivities. The study will be planned in accordance with the procedure reported in the literature.
7.4 HAS ANIMAL ETHICAL COMMITTEE CLEARANCE BEEN OBTAINED FROM YOUR INSTITUTION IN CASE OF 7.3?
The copy of the ethical clearance of certificate is obtained from Institutional Animal Ethical Committee (IAEC).
REFERENCES:
  1. Williams DA, Lemke TL. Non steroidal Anti-inflammatory Drugs. Foye’s principles of medicinal chemistry. 5thed. International Student Edition by Lippincott Williams and Wilkins; 2002. p. 751-93.
  2. Frank MM, Fries LF. The role of complement in inflammation and phagocytosis. Immunol Today 199;12:322.
  3. Vane J, Botting R. Inflammation and the mechanism of action of anti-inflammatory drugs. The Faseb J 1987;11:89-96.
  4. Jason B, Chris M. Natural approaches to controlling inflammatory diseases, natural anti-inflammation supplements and herbal treatments. Inflamm res 1998;47:334-8.
  5. Arthritis. [online]. 2011 Feb 14; Available from:URL:
  6. Bendele AM. Animals models of rheumatoid arthritis. J Musculoskel Neuron Interact 2001;1(4):377-85.
  7. Wooley PH. What Animal Model are Best to Test Novel Rheumatoid Arthritis Therapies. CurrRheumatol Rev 2008;4:277-87.
  8. Warrier PK and Ramankutty C. Indian Medicinal Plants. Delhi 1: Orient Longman; 1994.p. 132-6.
  9. Shankhajit DE, Dey YN, Ghosh AK. Anti-Inflammatory Activity of Methanolic Extract of Amorphophallus Paeoniifolius and its Possible Mechanism. Int J Pharm Bio sci 2010;1(3):1-8.
  10. Purwal L, Shrivastava V, Jain UK. Studies on Anti-Diarrhoeal Activity of Leaves of Amorphophallus Paeoniifolius in Experimental Animals. Int J of Pharm Sci Res 2011;2(2):468-71.
  11. Nataraj HN, Murthy RLN, Setty SR. In Vitro Evaluation of Gastro-Protective of Suran-A Possible Explanation Through HPTLC Analysis. Int Res J Pharmacy 2011;2(9):103-6.
  12. Dey YN, Ghosh AK. Evaluation of Anthelmintic Activity of the Methanolic Extract of Amorphophallus Paeoniifolius Tuber. Int J Phar Sci Res 2010;1(11):117-21.
  1. Yoganarasimhan SN. Medicinal Plants of India. Bangalore: Interline Publishing Pvt Ltd; 1996. vol 1 p. 35.
  2. Chau TT. Analgesic testing in animal models. In: Pharmacological methods in the control of inflammation. Alan R Liss Inc 1989; p. 195-212.
  3. Agarwal RB, Rangari VD. Antiinflammatory and antiarthritic activities of lupeol and 19a-h LUPEOL isolated from Strobilanthus callosus and Strobilanthus ixiocephala roots. Ind J Pharmacol 2003;35:384-7.
  4. Bhardwaj KL, Patil KS, Kaushik M, Sahu A, Prakash Y, Verma KV. Study on efficacy of treatment with Ficus benghalensisleaf extracts on freund’s adjuvant induced arthritis in rats. Int J Drug Dev Res 2010; 2(4):3-6.

9. / SIGNATURE OF THE STUDENT
10. / REMARK OF THE GUIDE
The above mentioned information and literature has been extensively investigated, verified and is found to be correct. The present study will be carried out under my supervision and guidance.
11. / 11.1 NAME AND DESIGNATION OF THE GUIDE
11.2 SIGNATURE / Dr. PREETI KULKARNI M.Pharm.,Ph.D.
PROFESSOR,
DEPT. OF PHARMACOLOGY,
SET’s COLLEGE OF PHARMACY S.R.NAGAR, DHARWAD- 580002
11.3 HEAD OF THE DEPARTMENT
11.4 SIGNATURE / Dr. A R.Kulkarni M.Pharm.,Ph.D.
PROFESSOR,
DEPT. OF PHARMACOLOGY,
Set’s College of Pharmacy
s.r.nagar, Dharwad-580002
12. / 12.1 REMARK OF THE PRINCIPAL / The above mentioned information is correct and I recommend the same for approval.
12.2 SIGNATURE / Dr. V H. Kulkarni M.Pharm.,Ph.D.
PROFESSOR AND PRINCIPAL,
Set’s College of Pharmacy
s.r.nagar, Dharwad-580002

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