RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
KARNATAKA, BANGALORE
M. PHARM SYNOPSIS
YEAR OF ADMISSION - MAY 2009
TITLE OF THE SYNOPSIS
Effect of Chromolaena odorata Linn. on
streptozotocin-induced diabetes and cataract in rats
BY
Mr. ONKARAMURTHY M
M.PHARM, PART-I
DEPARTMENT OF PHARMACOLOGY
UNDER THE GUIDANCE OF
Mr. SHIVALINGE GOWDA K P M.Pharm., DPPM., (Ph.D).
Asst. Professor
DEPARTMENT OF PHARMACOLOGY
INSTITUTION
SREE SIDDAGANGA COLLEGE OF PHARMACY
B.H. ROAD, TUMKUR-572102.
KARNATAKA
ANNEXURE - II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. / Name of the Candidateand Address / Onkaramurthy M
M Pharm, Part -1
Department of pharmacology
Sree Siddaganga College of Pharmacy
B.H. Road, Tumkur-572102
2. / Name of the Guide / Mr. Shivalinge Gowda K P M.Pharm.,DPPM., (Ph.D).
Asst. Professor
Department of pharmacology
Sree Siddaganga College of Pharmacy
B.H. Road, Tumkur-572102
3. / Name of the Institution / Sree Siddaganga College of Pharmacy
B.H. Road, Tumkur-572102
4. / Course of Study and Subject / M Pharm – (Pharmacology)
5. / Date of Admission / 23/05/2009
6. Title of the Topic:
Effect of Chromolaena odorata Linn. on
streptozotocin-induced diabetes and cataract in rats
7.0 /
BRIEF REVIEW OF THE INTENDED WORK
7.1 INTRODUCTION
Diabetes mellitus is a metabolic disorder characterized by hyperglycemia, glucoseurea and negative nitrogen balance and it is mainly due to lack of insulin secretion in beta cells of pancreas and desensitization of insulin receptors for insulin.1 Defective insulin secretion is the major cause for chronic hyperglycemia resulting in impaired function or failure of various organs such as eyes, kidneys, nerves, heart and blood vessels.2 Diabetes mellitus is one of the common metabolic disorder with micro and macro vascular complications that results in significant morbidity and mortality. It is considered as one of the five leading causes of death in the world.3
Type 1 diabetes mellitus is characterized by loss of the insulin-producing beta cells of the islets of langerhans in the pancreas, leading to a deficiency of insulin. The main cause of this beta cell loss is a T-cell mediated autoimmune attack.4 Type2 diabetes mellitus is characterized differently due to insulin resistance or reduced insulin sensitivity, combined with reduced insulin secretion. Obesity is found in approximately 55% of patients diagnosed with type2 diabetes.5
Cataract is characterized by cloudiness or opacification of the eye lens is the leading cause of blindness all over the world. In view of the widespread prevalence of diabetes in developing countries like India, diabetic cataract may pose a major problem in the management of blindness.
Although the pathogenesis of diabetic complications is not known, many biochemical pathways associated with hyperglycemia have been implicated. Among these, the polyol pathway has been extensively studied. Aldose reductase (AR) is the first and rate-limiting enzyme of the polyol pathway. Under euglycemic conditions, AR plays a minor role in glucose metabolism during diabetes, its contribution is significantly enhanced leading to a conversion of excess glucose to sorbitol in insulin independent tissues like the lens. AR dependent synthesis of excess polyols has been implicated as one of the mechanisms leading to diabetic cataract.6
7.2 NEED FOR THE STUDY
In the present situation, diabetes is possibly the world’s largest growing metabolic disorders and as the knowledge on the heterogencity of this disorder is advanced, the need for more appropriate therapy increases. A number of allopathic drugs are used for the antidiabetic effect like tolbutamide, metformin, phenformin, and acarbose has danger of drug interaction and adverse effect etc.7 Traditional plant medicines are used throughout the world for a range of diabetic presentations. There are many medicinal plants known to be used in the treatment of diabetes and a number of plants had been screened positive for their antidiabetic effect.8
According to the International Diabetes Federation, in 2007, it is revealed that the country with the largest numbers of people with diabetes is India (40.9 million), followed by China (39.8 million), the United States (19.2 million),Russia (9.6 million) andGermany (7.4 million) by the year 2025, there will be as many as 7 million new diabetic cases in the world.9
Cataract is a major health problem and major cause of blindness throughout the world. Currently, the only available treatment for the disease is the surgical replacement with a synthetic implant. Although such a surgical replacement of the natural lens with an artificial lens is significantly effective in restoring vision to most patients, it is not free of complications. Attempts to prevent cataract formation, or at least significantly retard the onset of the disease would be of great value.10
It is predicted the current 20 million people in the world with severely reduced visual acuity by the cataract is going to swell to 40 million by the year 2020.11
Some of the Ayurvedic plants have been proven to be effective in the diabetic cataract. Hence the interest developed in this field.12
Chromolaena odorata Linn. is in use amongst the natives of India in diabetes13 and also used in eye problem in various countries.14 Therefore the present study is aimed at elucidating the antidiabetic and prevention of diabetic cataract profile of Chromolaena odorata Linn.
7.3 REVIEW OF LITERATURE
Description: A herb usually growing as an invasive plant. Leaves are simple,
opposite and dentate, flowers are whitish.
Classification
Kingdom : Plantae
Phylum : Tracheophyta
Class : Magnoliopsida
Order : Asterales
Family : Asteraceae
Subfamily : Asteroideae
Tribe : Eupatorieae
Genus : Chromolaena
Botanical name : Chromolaena odorata Linn.
Synonyms
1. Eupatorium odoratum Linn.
2. Osmia odorata Linn.
Common names :
English :Triffid weed, bitter bush, chromolaena, jack in the bush, siam
weed, christmas bush.
Kannada :Kamyunist kale
Tamil :Vettukayapachai
Assamese :Jarmaniban
Mizoram : Tlangsam.
Distribution: India, China, Indonesia, East Timor, Philippines, Tropical America
and South Africa.15 16
Chemical constituents: Alkaloids, saponins, tannins, phlobatannins, anthraquinones,
steroids, terpenoids, flavonoids and cardiac glycosides.17
Parts used: Leaves and flowers.
Ethnomedical uses: In traditional medicine, the leaves are used for wound dressing,
skin infection and to stop bleeding, as an antispasmodic, antiprotozoal, antifungal
antitrypanosomal, antibacterial, , antihypertensive, anti-inflammatory, astringent,
diuretic and hepatotropic agent.17
Reported activities: Antipyretic, anti-inflammatory, analgesic,18 wound healing,18
Antiandrogenic,19 antifunga,l20 antimycobacterial,21 antiprotozoal,22 immuno-
modulatory23 and antioxidant.17
7.4 OBJECTIVES OF THE STUDY
- Authentication and collection of plant material.
- Alcoholic extraction of plant material by soxhlet process.
- To investigate the effect of Chromolaena odorata Linn. on streptozotocin induced diabetes and cataract in rats
8.0
/ MATERIALS AND METHODS
8.1 Collection and extraction
The leaves of Chromolaena odorata Linn. will be collected and shade dried. The
powdered dried leaves will be extracted with alcohol by soxhlet process.
8.2 Experimental design
Experimental animals: Male albino rats weighing approximately 150-200 g will
be used. The animals will be fed with standard chow and will be given water ad
libitum.
Dose: A dose of 200 mg/kg body weight per oral (b.w.p.o) of alcoholic extract of
Chromolaena odorata Linn. (ACO) will be taken as per the work.24
1.Hypoglycemic activity 3
Grouping of animals: The animals are divided into three groups of six rats in each group.
Group / Treatment / Dose
(mg/kg) / ROA / parameters
1
2
3 / Normal Control (NS)
ACO
Glibenclamide / ------
200
0.5 / p.o
p.o
p.o / 1) Blood glucose level.
2) Body weight.
3) Glutathione.
4) Thiobarbituric acid reactive
substances (TBARS).
5)Superoxide dismutase (SOD).
6) Catalase.
Where ROA-route of administration, ACO-Alcoholic extract of Chromolaena odorata
Linn. NS-Normal saline
Duration of the study: The study will be conducted for 15 days.
Short term studies: Animals were fasted for 16 h prior to drug administration
allowing access only to water. Blood samples will be collected from retro-orbital
plexus of each rat at 0, 30, 60, 120 min. The samples will be analyzed for blood
glucose content by using glucose-oxidase/peroxidase method.
Long term studies: Blood samples were collected from the overnight fasted
animals and fasting blood glucose levels will be measured at 0, 5, 10, 15 day in the
study and samples will be analyzed for blood glucose content by using glucose-
oxidase/peroxidase method. On 15th day, the blood samples will be studied for SOD
TBARS, glutathione, catalase. Statistical analysis will be done by ANOVA.
2. Diabetic cataract activity 1, 6 ,11, 24, 25
A. Experimental animals: Male albino rats weighing approximately 150-200 g will be used. The animals will be fed with standard chow and will be given water ad libitum.
B. Dose: A dose of 100 mg/kg and200 mg/kg body weight per oral (b.w.p.o) of alcoholic extract of Chromolaena odorata Linn. (ACO) will be given.24
C. Grouping of animals: The animals are divided into five groups of six rats in
each group.
All animals except normal control group will be injected Streptozotocin 65 mg/kg
b.w.i.p.25
Duration of the study: The diabetic study will be conducted for a period of 15 days
and it will be continued for another 6 weeks for cataract study. The dose will be
administered once daily until the study is complete.
D. Experimental design
Group / Treatment / Dose
(mg/kg) / ROA / parameter
1
2
3
4
5 / Normal control(NS)
Diabetic control(DC)
DC+ACO
DC +ACO
DC+ glibenclamide / ------
65
100
200
0.5 / p.o
i.p
p.o
p.o
p.o / 1) Blood glucose level.
2) Body weight.
3) Glutathione.
4) Thiobarbituric acid reactive
substances (TBARS).
5) Superoxide dismutase (SOD).
6) Catalase.
7) Cataract score.
NS-Normal saline, ROA-route of administration, ACO-Alcoholic extract of Chromolaena odorata Linn.
Short term studies: Animals were fasted for 16 h prior to drug administration
allowing access only to water. Blood samples will be collected from retro-orbital
plexus of each rat at 0, 30, 60, 120 min. The samples will be analyzed for blood
glucose content by using glucose-oxidase/peroxidase method.
Long term studies: Blood samples will be collected from the overnight fasted
animals and fasting blood glucose levels will be measured at 0, 5, 10, 15 day in the
study and samples will be analyzed for blood glucose content by using glucose-
oxidase/peroxidase method. On the 15th day blood samples will be analysed for
SOD, TBARS, glutathione, catalase. Statistical analysis will be done by ANOVA.
Cataract score is measured by using slit Lamp, Where cataract score and blood
glucose level will be measured once in a week upto 8 weeks.
8.3 DOES THE STUDY REQUIRE ANY INVESTIGATION OR INTERVENTIONS TO BE CONDUCTED ON PATIENTS OR OTHER HUMANS /ANIMALS? IF SO PLEASE DESCRIBED BRIEFLY.
Yes, the above study requires investigation to be done on the albino Wistar rats for the determination of diabetes and cataract.
8.4 HAS ANIMAL ETHICAL COMMITTEE CLEARANCE BEEN OBTAINED FROM YOUR INSTITUTION IN CASE?
The study has been referred to the ethical committee of the institution for clearance. (Approval no: 80/2009-10, Date :30-11-2009) Enclosed copy annexure I
9 / References:
1. Selvan VT, Manikandan L, Senthil Kumar GP, Suresh R, Kakoti BB, Gomathi P et al. Antidiabetic and antioxidant effect of methanol extract of Artanema sesamoides in streptozotocin induced diabetic rats. Int J Appl Res Nat Products 2008;1:25-33.
2. Susheela P, Balaravi P, Theophilus J, Reddy TN, Reddy PU. Evaluation of hypoglycemic and antidiabetic effect of Melia dubia CAV fruits in mice. Curr Sci 2008;94:91-5.
3. Semwal BC, Shah K, Chauhan NS, Badhe R, Divakar K. Antidiabetic activity of stem bark of Berberis aristata D.C. in alloxan induced diabetic rats. Int J Pharmacol 2008;6:1-6.
4. Rother KI. Diabetes treatment-bridging the Divide. N Engl J Med 2007;356:1499–501.
5. Valeriya L, Anna J, Peter A, Nicolo P, Tiinamaija T. Clinical risk factors, DNA variants, and the development of type 2 diabetes. N Engl J Med 2008;359:2220–32.
6. Surayanarayana P, Saraswat M, Petrash JM, Reddy GB. Emblica officinalis and its enriched tannoids delays streptozotocin induced diabetic cataract in rats. Mol Vis 2007;13:1291-7.
7. Tripathi KD. Essentials of medical pharmacology. 5th ed. New Delhi: Jaypee Brothers Medical Publishers (P) Ltd;2003.
8. Modak M, Dixit P, Londhe J, Ghaskadbi S, Devasagayam TP. Indian herbs and herbal drugs used for the treatment of diabetes. J Clin Biochem Nutr 2007;40:163–73.
9. Diabetes statistics and Death Rates. Available from: http://health.savvy-cafe.com/diabetes-statistics-india-is-the-diabetic-capital-of-the-world.(accessed on 25 Oct 2009).
10. Javadzadeh A, Ghorbanihaghjo A, Bonyadi S, Rashidi MR, Mesgari M, Rashtchizadeh N et al. Preventive effect of onion juice on selenite induced experimental cataract. Indian J Opthalmol 2009;57:185-9.
11. Saito M, Ueo M, Kametaka S, Saigo O, Ushida S, Hosaka H et al. Attenuation of cataract progression by A-3922, a dihydrobenzofuran derivative, in streptozotocin- induced diabetic rats. Biol Pharm Bull 2008;31:1959-63.
12. Rathi SS, Grover JK, Vikrant V, Biswas NR. Prevention of experimental diabetic cataract by Indian ayurvedic plant extracts. Phytother Res 2002;16:774-7.
13. Pullaiah T, Naidu KC. Antidiabetic plants in India and herbal based antidiabetic research 2003; 6:135-6.
14. The Status of Chromolaena odorata (L.) R.M. King and H. Robinson in West and Central Africa. Avaliable from: http://www.ehs.cdu.au/ chromolaena /proceedings /third /3boob.html. (accessed on 5 Nov 2009).
15. Chromolaena odorata(Jack in bush). Avaliable from: http://zipcodezoo.com/Plants /C/ Chromolaena_odorata/Default.asp. (accessed on 15 Nov 2009).
16. Chromolaena odorata(herb). Availablefrom: http://www.issg.org/database/ species/ references.asp?si=47&fr=1&sts=&lang=EN. (accessed on 15 Nov 2009).
17. Afolabi C, Moladan A, Ibukun EO, Dan-ologe IA. Phytochemical constituents and antioxidants properties of extract from the leaves of Chromalaena odorata. Sci Res Essays 2007;2:191-4.
18. Owoyele BC, Oguntoye SO, Dare K, Ogunbiyi BA, Aruboula EB, Soladoye AO. Analgesic , anti-inflamatory and antipyretic activities fractions from Chromolaena odorata. J Med Plant Res 2008;2:219-25.
19. Yakubu MT, Akanji MA, Oladiji AT. Evaluation of antiandrogenic potentials of aqueous extract of Chromolaena odoratum (L.) K. R. leaves in male rats. Andrologia 2007;39:235-43.
20. Ngono NA, Ebelle ER, Ndifor F, Biyiti L, Amvam ZP, Bouchet P. Antifungal activity of Chromolaena odorata (L.) King & Robinson (Asteraceae) of Cameroon. Chemother 2006 ;52:103-6.
21. Apichart S, Apinya C, Tananit S, Somnuk B1, Puntip Timsuksai2 Saovaluk V et al. Antimycobacterial activity and cytotoxicity of flavonoids from the flowers of Chromolaena odorata. Arch Pharm Res 2004;27:507-11.
22. Taleb-Contini SH, Salvador MJ, Balanco JM, Albuquerque S, Oliveira DC. Antiprotozoal effect of crude extracts and flavonoids isolated from Chromolaena hirsuta (asteraceae). Phytother Res 2004;18:250-4.
23. Taleb-Contini SH, Kanashiro A, Kabeya LM, Polizello AC, Lucisano-Valim YM, Oliveira DC. Immunomodulatory effects of methoxylated flavonoids from two Chromolaena species: structure-activity relationships. Phytother Res 2006;20:573-5.
24 Owoyele VB, Adediji JO, Soladoye AO. Anti-inflammatory activity of aqueous leaf
extract of Chromolaena odorata. Inflammopharmacol 2005;13:479-84.
25 Bajaj S, Srinivasan BP. Investigations into the anti-diabetic activity of
Azadirachta indica. Indian J Pharmacol 1999;31:138-41.
9 / SIGNATURE OF THE CANDIDATE
10 / REMARK OF THE GUIDE:
The above information and literature has been extensively investigated, verified and was found to be correct. The present study will be carried out under my supervision and guidance.
11 / 11.1 NAME AND DESIGNATION
OF GUIDE / Mr. Shivalinge Gowda K P
M.Pharm.,DPPM., (Ph.D).
Asst. Professor
Department of pharmacology
Sree Siddaganga College of Pharmacy
B.H. Road, Tumkur-572102
11.2 SIGNATURE
11.3 CO-GUIDE /
NA
11.4 SIGNATURE / NA
12 / 12.1 HEAD OF THE DEPARTMENT / Dr. THIPPESWAMY B.S. M.Pharm., Ph.D
Professor & Head of the Department
Dept.of pharmacology,
Sree Siddaganga College of Pharmacy,
B.H.Road, Tumkur-572 102
12.2 SIGNATURE
13 / 13.1 REMARKS OF THE PRINCIPAL / The above mentioned information is correct and I recommend the same for approval.
13.2 SIGNATURE / Dr. S.BADAMI M.Pharm., Ph.D
Principal
Sree Siddaganga College of Pharmacy
B.H. Road, Tumkur-572102
3