EVALUATION OF ANTI-ARTHRITIC ACTIVITY OF AERIAL PARTS
OF Justicia tranquebariensis LINN. IN WISTAR RATS.
SYNOPSIS FOR
M. PHARM DISSERTATION
SUBMITTED TO
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
KARNATAKA, BANGLORE
BY
SHRINIVAS S SHAIWALE
M. PHARM PART-1
DEPARTMENT OF PHARMACOLOGY
NARGUND COLLEGE OF PHARMACY
BANGALORE-560085
(2010-2012)
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES BANGALORE, KARNATAKA.
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR PG DISSERTATION
1. / Name of the candidate and address(in block letters) / SHRINIVAS S SHAIWALENARDUND COLLEGE OF PHARMACY,
DATTATREYA NAGAR, II MAIN,100FT RING ROAD,BSK III STAGE,
BANGALORE-560085
2. /
Name of the institution
/NARGUND COLLEGE OF PHARMACY
DATTATREYA NAGAR, II MAIN,100FT RING ROAD,BSK III STAGE,BANGALORE-560085
3. /
Course of study and subject
/MASTER OF PHARMACY IN PHARMACOLOGY
4. /Date of the admission
/ 06/07/20105. / Title of the topic :
EVALUATION OF ANTI-ARTHRITIC ACTIVITY OF AERIAL PARTS
OF Justicia tranquebariensis LINN. IN WISTAR RATS.
6.
7.
8. / BRIEF RESUME OF THE INTENDED WORK
6.1 NEED FOR THE STUDY:
“Public health in the future will be increasingly about increasing the quality of life, not merely its length. Arthritis, with the pain and limitation it inflicts on millions of our people, young and old, sits right in the center of that future.”- James Marks, senior vice president, Robert Wood Johnson Foundation.
Arthritis is a term used to describe more than 100 different conditions that affect joints as well as other parts of the body. Arthritis is one of the most prevalent chronic health problems and one of the nation’s most common causes of disability. Over the next 25 years as the Baby Boom generation continues to age, the toll of this disease will escalate. To have a clear picture of the looming disease burden and its impact on our nation's health care and public health systems, estimating disease prevalence, the number of people affected by any form of arthritis is critical.1 Arthritis can interfere with a person’s ability to work and be productive. It is currently the third leading cause of work limitation in the United States. Physical inactivity contributes to the pain, dysfunction, and disability associated with arthritis and also contributes to poor outcomes for people with diabetes, heart disease, and other chronic conditions. Being physically active is important for people with arthritis because it helps them manage pain and maintain function and mobility; it is also important for helping them control their weight, lessen their chance of a fall, and reduce their risk of depression and other chronic conditions.2
Rheumatoid arthritis is a chronic inflammatory disease involving multiple joints. The main pathology of affected synovial tissue consists of hyperplasia and subintimal infilteration of T and B lymphocytes. Synovial tissue hyperplasia forms the pannus tissue that irreversibly destroys the cartilage and bone in the affected joint. It affects the people in their prime of life between the age of 20-50 years with unpredictable course can leads to physical disability in some. It is a common disorder with clinical signs and symptoms related multiple anatomical sites, both articular and extra-articular. Presently, the non steroidal anti-inflammatory drugs (NSAIDs) are the main stay in this condition, however, they have serious adverse effects and have limitations for a long term therapy. The immunosuppressive drugs are reserved for selected cases, while the disease modifying drugs like gold-salts are costly and have low benefit risk ratio. Hence, there is a need for drugs having good efficacy with low toxic profile anti-oxidant property in this debilitating disorder.3
From the literature survey it is evident that some species of genus Justicia have effective anti inflammatory4 and antiarthritic5 activity. Although mention is made in the literature regarding the usefulness of Justicia tranquebariensis as an antibacterial, antipyretic and antioxidant there is no report of it being tried as an antiarthritic agent. The current study is being carried out to investigate for the presence of antiarthritic property.
6.2 REVIEW OF LITERATURE:
Justiciatranquebariensis is used in Ayurveda, Folk and Sidha. Some species of genus Justicia have been used in traditional system of medicine for the treatment of fever,6 pain,7 inflammation,8 diabetes,9 diarrhoea,10 liver disease,11 analgesic and anti-inflammatory4 activities.
Scientific name: Justiciatranquebariensis
Synonyms: Pindi (Sanskrit), Sivanarvembu (tamil) Kaddiyarakina (kannada) Genus: Justicia
Species: tranquebariensis
Family: Acanthaceae
Order: Scrophulariales
Botanical Description:
Justiciatranquebariensisis a small shrub, which is widely distributed in southern parts of India (Deccan, Mysore southwards). The plant has a place amongst many others in the Hortus Bengalensis, and is common on the Coromandel coast. It would appear to have been first particularly observed by Koeing, near Tranquebar, and is perhaps, the Justicia pervifolia of Lamarck.12,13
The plant has following parts :
Roots; perennial, somewhat granulated
Stems; in young plants erect, in old ones uncertain
Branches; annual, small, staggling, round, a little downy, jointed & swelled above the joints, from 1-2 ft long
Leaves; opposite, short petioled, oval, round, entire, pretty smooth
Spikes; termina, penducled, long, erect, generally four-sided occasioned by four rows of ovoicular, short-petioled, opposite, ciliate breactres.
Flowers; a mixture of pale yellow, and red; standing in the axills of the two rows of bracets that occupy forepart of the spikes; on the back part they are generally wanting.
Flowering time the wet, and cold season.14
Plant chemicals:
The Justiciatranquebariensis plant parts yielded several lignans, phytosterols, brassicasterol, campesterol, ergostadienol, stigmasterol, sitosterol, spinasterol, isofucostil and a sterol glucoside, beta-sitosterol-Oglucoside.15
Uses:
The juice of small and somewhat fleshy leaves of this plant is considered by the natives of India, as cooling and aperients, and is prescribed for children in small-pox.
Bruised leaves are also applied to blows and other external injuries, contusions.12,13
Young leaves are crushed and filtered in pure water, kept in dark for overnight, the infusion is given internally in the morning for curing jaundice.16
Justicia tranquebariensis also used in skin diseases & stomach problems.17
For Poisonous Bites: Handful of leaves, made into juice, taken internally for 3 days. Paste made of the leaves applied externally on the swelling to reduce the pain.18
Leaf juice applied for cut wounds and in the eyes to cure eye infection.19
M. shabana begum et al,(2009) investigation reports confirmed the therapeutic potency of Justicia tranquebariensis, and suggests that the plant extracts possess certain constituents with antibacterial properties that can be used as antimicrobial agents in new drugs for the therapy of infectious diseases caused by pathogens.20
M. shabana begum et al,(2009) reported the antipyretic activity of leaves of Justicia tranquebariensis in albino mice.21
G. Balmurgan et al,(2008) reported antibacterial and antifungal activities of chloroform and ethanolic extracts of aerial portions of Justicia tranquebariensis Linn by agar disc diffusion method. Also indicates in vitro antioxidant properties of the ethanolic extract of plant by free radical scavenging activity.22
6.3 OBJECTIVE OF THE STUDY:
The main objectives of the studies are :
1. Preparation of Justicia tranquebariensis aqueous and methanolic extracts
2. Preliminary phytochemical investigation of Justicia tranquebariensis
3. To perform acute toxicity studies as per OECD guidelines.
4. In vivo anti-arthritic activity will be studied by using Freund’s adjuvant induced arthritis in rats
5. In vivo anti-arthritic activity will also be studied by using Formalin induced arthritis in rats
6. In vitro anti-oxidant activity will be studies by using Superoxide scavenging, Hydroxyl scavenging activity.
MATERIAL AND METHOD
7.1 SOURCE OF DATA :
Data will be obtained from the experimental work, which include laboratory based animal studies
and parameters evaluated.
7.2 METHOD OF COLLECTION OF DATA
The data collected is based on animal experimentation as per the parameters studied under each
animal model. The sampling procedure and collection of data will be as follows.
Web sites:
· www.sciencedirect.com
· www.pubmed.com
· www.google.com
· www.ijp-online.com
· www.rjptonline.org
METHODOLOGY :
7.2.1 Acute Toxicity Studies:
Acute toxicity studies will be performed according to OECD guidelines. Based on the study two dose will be selected.
7.2.2 Freund’s adjuvant induced arthritis in rats23
Albino rats Wistar strain weighing between 150-200 g will be divided into 7 groups of 6 animals each, with 3 animals housed in each cage. Except those in the control group rats in all other groups are injected with, s.c 0.1 ml of Complete Freund’s Adjuvant into the planter region of the left hind paw.
Animals:
Species / Rat
Strain / Wistar
Age and sex / 4-6 weeks, Male
Body weight / 150-200 g
No. of animals in each group / N=6
No. of groups / Seven
Vehicle of herbal drugs / Distilled water
Vehicle of standard drugs / Distilled water
Water and food / Ad. Libitum
Groupings:
Group 01:Not administered Complete Freund’s Adjuvant (Normal control)
Group 02: Administered with only CFA (Arthritic control)
Group 03: Administered with CFA + Indomethacin ( 3mg/kg )24
Group 04: Administered with CFA + Justicia tranquebariensis aqueous extract low dose.
Group 05: Administered with CFA + Justicia tranquebariensis aqueous extract high dose.
Group 06: Administered with CFA + Justicia tranquebariensis methanolic extract low dose.
Group 07: Administered with CFA + Justicia tranquebariensis methanolic extract high dose.
The changes in the paw volume will be measured on various days upto 21 days following Freund’s adjuvant injection. The extracts and indomethacin will administered orally for 14 days from the day of Freunds adjuvant injection. On the 21st day rats will be anaesthized with ketamine and diazepam and various parameters estimated.
Ø Biochemical parameters to be estimated :
· Alkaline phospatase
· Serum glutamic oxaloacetate transaminase
· Serum glutamic pyruvate transaminase
Ø Histopathological parameters
7.2.3 Formalin induced arthritis inflammation25 :
Male Wistar rats (150-190 g) are divided into six groups of five animals each. Inflammation was produced by subplantar injection of 0.1 mL of 2 % v/v formalin in normal saline in the right hind
paw of the rats on the first and third day. The extract (low and high doses) and distilled water (3 mL/kg)
to be administered orally once a day for 10 days. Indomethacin (5 mg/kg) given orally, is used as
standard. The rat paw thickness is measured daily for 10 days. The percentage inhibition of the mean increase in the paw edema of each group is to be calculated on the tenth day and compared with the control.
Group 01: Arthritic control (2% v/v formalin)
Group 02: Arthiritic rats will be administered Indomethacin
Group 03: Arthiritic rats will be administered Justicia tranquebariensis aqueous extract low dose.
Group 04: Arthiritic rats will be administered Justicia tranquebariensis aqueous extract high dose.
Group 05: Arthiritic rats will be administered Justicia tranquebariensis methanolic extract low dose.
Group 06: Arthiritic rats will be administered Justicia tranquebariensis methanolic extract high dose.
7.2.4 In vitro antioxidant activity of Justicia tranquebariensis :
A) Superoxide scavenging activity26
B) Hydroxyl scavenging activity26
Statistical analysis:
The results are expressed as mean ± SEM will be calculated for each parameter. For the determinations for significant inter group differences each parameters will be analysed separately by one way analysis of variance (ANOVA) and post Hock analysis will be done by Dunnett’s test.
7.3 Does the study require any investigation to be conducted on patients or other humans or animals? If so please describe briefly.
Yes, the above study involves an investigation on whole animals and also on their internal organs. The test animals namely the Wistar rats will have to be sacrificed to evaluate the end point indicators such as Bone joint histopathology.
7.4 Has ethical clearance been obtained from your institution?
Applied for the IAEC of Nargund college of pharmacy
List of References
1. Chopra A, Saluja M, Patil J, Tandale HS. Pain and Disability, Perceptions and Beliefs of a Rural Indian Population: AWHO-ILAR COPCORD Study. The J of Rheumatology 2002;29(3):614-21.
2. Theis KA, Murphy L, Hootman JM, Helmick CG, Yellin E. Prevalence and correlates of arthritis-attributable work limitation in the US population among persons 18-64: 2202 National Health Data Survey. Arthritis Rheum 2007;57(3):355-63.
3. Kulkarni RR, Patki PS, Jog VP, Gandage, Pathwardhan B. Efficacy of an ayurvedic formulation in rheumatoid arthritis: A double blind, placebo controlled, cross-over study. Indian J Pharmacol. 1992;24:98-101
4. Lino CS, Taviera ML, Viana GSB, Matos FJA. Analgesic and anti-inflammatory activity of Justicia pectorails Jacq and its main constituents: coumarin and umbelliferone. Phytotherapy Res 1997;11(3):211-5
5. Paval J, Kaitheri SK, Potu BK, Govindan S, Kumar RS, Narayanan SN et al.Anti-arthritic potential of plant Justicia gendarussa Burm F. Clinics 2009;64(4):357-60.
6. Chen CC, Hsm WC, Ko FN, Hyang YL, Ou JC, Teng CM. Antiplatelet arylnaphthalide lignans from Justicia procumbens. J Nat Prod 1996;59:1149
7. Bhattarai NK. Medical ethnobotany in the raptizone. Nepal Filoterapa 1993;64:483
8. Panthong A, Kamjapothin D. Ethanobotanical review of medicinal plants from Thai traditional books. Part 1: Plants with anti inflammatory properties. J Ethnopharmacol 1986;18:213.
9. Mahabir D. Use of medicinal plants for diabetes in Trinidad and Tobago Rev planum salud publ/PAN. Am J Pub Health 1997;1:174
10. Heinrich M, Kuhnt M, Wright CW, Rimplex H, Pillipson JD, Schandelmaier A, Warhurst DC. Parasitological and microbiological evaluation of Mexic Indian medicinal plants. J Ethanopharmacol 1992;36:81
11. Yangfg LL, Yen KY, Kiso V, Kikino H. Antihepatotoxic actions of Formosan plant drugs. J Ethanopharmacol 1987;19:103.
12. Whitelaw Ainslie. Materia indica. London(UK): A & R Spottiswoode;1826. vol 2. p. 412
13. Nadkarni KM, Nadkarni AK. Indian materia medica. 3rd ed. Mumbai: Populalar Prakashan; 2007. vol 2. p. 715
14. William Roxburgh. Flora indica or description of Indian plants. William Carey, editor. Serampore(West Bengal):Mission Press;1820. vol 1. p. 131.
15. Khare CP. Indian medicinal plants. New York(NY): Springer Reference; 2007. p. 350
16. Senthilkumar M, Gurumoorthi P, Janardhanan K. Some medicinal plants used by Irular, the tribal people of Marudhamalai hills. Coimbatore, Tamil Nadu. Natural Product Radiance 2006;5(5): 382-8.
17. Ganesan S, Ponnuchamy M, Kesavan L, Selvaraj A. Floristic composition and practices on the selected sacred groves of pallapathy village (Reserved forest), Tamil Nadu. Indian J Trad Knowledge 2009;8(2):154-62
18. Sandhya B, Thomas S, Isabel B, Shenbagarathai R. Ethnomedicnal plants used by the valaiyan community of piranmalai hills (reserved forest), Tamilnadu, India. Afr J Trad CAM 2006;3(1):101-14
19. Ganesan S, Venkateshan G, Banumathy N. Medicinal plants used by ethnic group thottianaickans of semmalai hills (reserved forest), Tiruchirapalli district, Tamil Nadu. Indian J Trad Knowledge 2006;5(2):245-52.
20. Begum MS, Ilyas MHM, Burkanudeen A.Evaluation of antibacterial activity of leaves of Justicia tranquebariensis Linn. The Pharmacist 2008;3(1):35-7.
21. Begum MS, Ilyas MHM, Burkanudeen A.Antipyretic activity of extract of leaves of Justicia tranquebariensis (Linn) in albino mice. The Pharmacist 2009; 4(1):49-51.
22. Balmurugan G, Arunkumar MP, Muthusamy P, Anbazhagan S. Preliminary phytochemical screening, free radical scavenging and antimicrobial activities of Justicia tranquebariensis Linn. Research J of Pharm and Technology 2008;1(2):116
23. Mahesh SK, Naik SR, Kohli RK. Anti-inflammatory, antiarthritic and analgesic activity of a herbal formulation (DRF/AY/4012). Indian J of Experimental Biology 2007;45:278-84.
24. Philippe L, Pascale GP, Guingamp C, Bordji K, Terlain B, Netter P et al. Relations between functional, inflammatory and degenerative parameters during adjuvant arthritis in rats. Am J Physiol Integr Comp Physiol 2007 Oct;273(4):1550-6.
25. Igbe I, Ching FP, Eromon A. Anti-inflammatory activity of aqueous fruit pulp extract of Hunteria umbellate K schum in acute and chronic inflammation. Acta Poloniae Pharmaceutica – Drug Research 2010;67(1):81-5
26. Rajesh K, Satish kumar BP, Harsha R, Dinesha R, Irfan AM, Thammana GSS et al. In vitro antioxidant activity of leaves of Mentha arvensis Linn. J Pharm Research 2010;3(8):1951-4.
9. / SIGNATURE OF THE CANDIDATE / SHRINIVAS S SHAIWALE
10. / REMARKS OF THE GUIDE / RECOMMENDED
11.
/ NAME AND DESIGNATION
11.1 GUIDE / Prof. D S PURANIK
ASST. PROFESSOR
DEPT. OF PHARMACOLOGY,
NARGUND COLLEGE OF PHARMACY, BANGALORE-560 085.
11.2 SIGNATURE
11.3 CO-GUIDE (IF ANY) / S B CHANDRASEKAR
RESEARCH OFFICIER
NATURAL REMEDIES PVT LTD.
VEERSANDRA INDL. AREA, BANGLORE
11.4 SIGNATURE / .
11.5 HEAD OF THE DEPARTMENT / DR. H.J.HRISHIKESHAVAN
PROFESSOR & H.O.D,
DEPT. OF PHARMACOLOGY,
NARGUND COLLEGE OF PHARMACY, BANGALORE-560 085.
11.6 SIGNATURE
12. / 12.1 REMARKS OF THE PRINCIPAL
/ RECOMMENDED FOR REGISTRATION
12.2 SIGNATURE
12.3 PRINCIPAL / Prof. D S PURANIK
PRINCIPAL,
NARGUND COLLEGE OF PHARMACY,
BANGALORE-560 085.
14