A STUDY ON ANTI-DIABETIC ACTIVITY OF FLOWER OF MUSA PARADISIACA ON STREPTOZOTOCININDUCED DIABETES MELLITUS

M. Pharm Dissertation protocol

Submitted to the

RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES,

BANGALORE, KARNATAKA

BY

DURGESH MOHAN SINGH

B.pharm

Under the Guidance of

Dr. V.N. BIRADAR

Professor,

Department of Pharmacology,

N.E.T.PharmacyCollege, Raichur

DEPARTMENT OF PHARMACOLOGY

N.E.T.PharmacyCollege

Raichur-584103

2007-2008

RAJIVGANDHIUNIVERSITY OF HEALTH SCIENCES,

BANGALORE, KARNATAKA

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / Name of the candidate and address: / DURGESH MOHAN SINGH
S/O-Shri Brij Bilash Singh
Moh-Bhairopur (Chhawni)
Post-Kunaraghat
Dist-Gorakhpur- 273008(UP)
2. / Name of the institution: / N.E.T.PharmacyCollege,
Raichur-584103.
3. / Course of study and subject: / Master of Pharmacy in Pharmacology
4. / Date of admission of course: / 21st May 2007
5. / Title of the topic: /

A STUDY ON ANTI-DIABETIC ACTIVITY OF FLOWER OF MUSA PARADISIACA ON STREPTOZOTOCIN INDUCED DIABETES MELLITUS

6. / Brief resume of the intended work:
6.1:Need for the study:
6.2:Review of the literature:
6.3:Main objective of the study: / ENCLOSURE-I
ENCLOSURE-II ENCLOSURE-III
7. / Materials and methods:
7.1:source of the data:
7.2:Methods of the collection of the data:
7.3: Does the study require any investigations or interventions to be conducted on patients other human or animals? If so, please describe briefly.
7.4:Has ethical clearance been obtained from your institute in case of 7.3 ? / ENCLOSURE-IV
ENCLOSURE-V
YES: (mice/rats)
YES:
IAEC NO: 576/2002/IAEC/CPCSEA
8. / List of references: / ENCLOSURE-VI

9.

/

Signature of the candidate:

/

(DURGESH MOHAN SINGH)

10.

/

Remarks of the guide:

/

The proposed work can be carried out in our

Laboratory at N.E.T.PharmacyCollege, Raichur.

11.

/

Name and designation of-

11.1Guide:
11.2 Signature:
11.3 Co-guide:
11.4 Signature:
11.5 Head of the department:
11.6Signature: / Dr. V.N.BIRADAR
Professor,
Department of Pharmacology, N.E.T.PharmacyCollege, Raichur
………
Mr. BHEEMACHARI
Assistant Professor,
Department of Pharmacology,
N.E.T.PharmacyCollege, Raichur.

12.

/

12.1 Remarks of the chairman

and principal:

12.2 Signature: / PROF. H. DODDAYYA
Principal,
N.E.T.PharmacyCollege,
Raichur

ENCLOSURE –I

6. Brief resume of the intended work:

6.1: Need for the study:

Diabetes mellitus is a syndrome associated with hyperglycemia, hyperlipidemia, oxidative stress, polyurea, polyphagia, polydypsia, ketosis, nephropathy, neuropathy and cardiovascular disorders1,2,3. As on date about 1.7% of the world population has been estimated to suffer from diabetes mellitus and is expected to rise to 3.6% by the year 20254.

In modern medicine no satisfactory effective therapy is yet available to cure diabetes mellitus. Though insulin therapy is used for the management of diabetes mellitus, there are several drawbacks like insulin resistance5, anorexia, nervosa, brain atrophy and fatty liver6.Requirement for refrigeration of the drug, skilled technicians and high cost, which are not affordable in poor economic community. Chronic treatment with sulfonylurea and biguanides are also associated with side effects7.

Herbals drugs have been usedmajor source for of treatment of diabetes mellitus and other disease since ancient time in India and rest of the world. In spite of tremendous strides in the modern medicine there is no effective remedy by which tight glycaemic control is possible without adverse affects8. Herbal drugs are mostly out of toxic or of less toxic with fewer side effects compared to the synthetic drugs. Hence, there is persistent interest all over the world to explore other alternative therapies like ayurveda, unani, homeopathy, siddha etc. which are believed to be effective, safer and economical.

In this context, upon extreme survey, it was found that. The flowers of Musa paradisiacahas been traditionally in the treatment of diabetes mellitus. Neverthless, its scientific study has not yet been reported. Hence in the present project, an effect has been made to evaluate the flower of Musa paradisiaca used for its antidiabetic activity in animal mode.

ENCLOSURE –II

6.2: Review of literature:

Musa paradisiaca (Banana) is a tree like herb (Family:Musaceae) with thick stem composed of convoluted leaf sheaths. Leaves are very oblong. It is distributed through out India and Malaysia. And belong to musaceae family. Flower of Musa sapientum (another species of banana) showed hypoglycemic activity on normal fasting rabbits10,11.The biological activities reported contained in the different parts of Musa paradisiaca are mainly the leaves19, roots25, fruits13, stem21,22 and seed24 However, no literature is available about any biological activitycontained in the flowers ofMusa paradisiaca.

Chemically Musa paradisiaca contains starch (unripe pulp fruit)12, iron, carbohydrates, flavonoids, biflavonoids, tannins, phenolic compounds9.

In folk medicine all parts of Musa paradisiaca are used to cure several disorders, such as:

Methanolic extract of mature, green fruits of Musa paradisiaca (MEMP) produced hypoglycemic activity in both normal and streptozotocin (STZ) induced diabetic mice using chlorpropamide as the reference antidiabetic agent13.

The different parts of Musa paradisiaca (fruit, pulp, leaves) have been used as a folk medicine for treatment of peptic ulcers14, analgesic15 and anti-asthmatic16.

Acitan, a natural product obtained from Musa paradisiaca stem and leaves produced antioxidant activity in vitro using commonly accepted assays17.

Fruit peel of Musa paradisiaca has been used for its antimicrobial activity against Bacillus subtilis, Staphylococcus aureus, Escheerichia coli, Pseudomonas aeruginosa, Candida albicans and Cryptococcus neoformansin comparisonwith the standards benzyl penicillin and streptomycin18.

In Nigeria all parts (especially leaves and roots) of Musa paradisiaca have been used in the management of sexual dysfunctions 19.

In Turkey all parts of Musaparadisiaca has been used for prevention of cancer20.

The stem juices of Musa paradisiaca have been reported for dissolving pre-formed stones and preventing the formation of stones in the urinary bladder of rats21, 22.

 Stem juice is used in nervous affectations like epilepsy, hysteria and in dysentery and diarrhea. Several oligosaccharides comprising fructose xylose, galactose, glucose and mannose occur naturally in banana, making it an excellent prebiotic for the selection growth of beneficial in the intestine 23.

Chhanda mallick et al24, have studied the separate and composite methanolic extract of seed of Eugenia Jambolana and Musa paradisiaca have some remedinal effects against streptozotocin-induced diabetic state.

Individual extract of root of Musa paradisiaca and leaf of Coccinia Indica produced antihyperglycemic effects in streptozotocin-induced diabetic male albino rate25.

ENCLOSURE-III

6.3: Main objective of study:

The main objective of the proposed work is to study the anti-diabetic activity of Musa paradisiaca extracts in STZ induced diabetic animals. According to the extensive traditional application in the ayurveda practice and the background of present literature on the plants (Musa paradisiaca), the present work is planned.

The present work is planned with the following objectives:

a) To prepare extract of Musa Paradisiaca flower.

b) Phytochemical investigation of the extracts.

c) To establish pharmacological activities of extracts.

d) Qualitative studies of the extracts.

ENCLOSURE-IV

7. Materials and methods:

7.1: Sources of the data:

Whole work is planned to generate data from laboratory based animal experimental studies as described in both the National and International journals and also from text books available with e-libraries, college, institutions and other reputed libraries.

The day today development in the area will be updated by conducting literature survey through e-publishing and Helinet provided by RGUHS, Banglore.

ENCLOSURE-V

7.2 Method of the collection of the data:

The whole study is divided into different phases:

Phase-I:

Collection of the drugs and preparation of the extracts24:

The flower of Musa paradisiaca is procured either from commercial herbal stores or from natural source. Fresh flowers of Musa paradisiacawill be cleaned, cut in to small pieces and shade dried or, in an incubator for 2-3 days at 400 C. Crushed it in an electric grinder and then powdered. Make extracts of powder by using different solvents. Extracts will be prepared by over night maceration and continuous hot extraction using Soxhlet apparatus.

Phase-II:

Preliminary phytochemical investigation:

The preliminary phytochemical investigation will be carried out with extracts of Musa paradisiaca for qualitative identification of phytochemical constituents present with extract. Test will be carried out by using standard methods26,27. All the chemicals and reagents proposed to be used will be of analytical grade.

Induction of diabetes25:

Fasting rats for 24hours, were subjected to single intramuscular injection of streptozotocin (STZ) at the dose of 4 mg/0.1 ml of citrate buffer/100 gm body weight/rat. Rats that exhibit blood glucose concentration more than 250mg/dl after 24hrs of STZ injection will be considered as diabetic and selected for the proposed study.

The blood glucose concentration before and after respective treatment at above specified time intervals will be determined by GOD/POD method.

Phase-III:

Pharmacological activities:

  1. Determination of Acute toxicity:

The acute toxicity of the aqueous extract of the flowers of Musa paradisiaca will be determined by using albino mice of either sex (20-25gms); those maintained under standard conditions. The animals will be fasted for 3hours prior to experiment. Animals will be administered with different doses of the extract by following up and down methods as per OECD guidelines number 42528. From LD50 dose 1/20, 1/10, and 1/5th doses are to be selected and will be considered as low, medium and high dose respectively.

B. Experimental methods:

1) Hypoglycemic activity:

This will be conducted for all the extracts proposed in the study. However, here the common procedure involved in the determination of hypoglycemic activity due to any extract is explained, which will be common for all other extracts.

Albino rats of either sex weighing between 150-200 gms will be categorized into five groups, each group consisting of 6 animals.

Group A:Normal control (volume of vehicle matched with the average volume of

total dose of the respective extract)

Group B:Standard (glibenclamide 90 µg/kg, p.o)

Group C:Extract of the flowers ofMusa paradisiaca(low dose).

Group D:Extract of the flowers of Musa paradisiaca (medium dose).

Group E:Extract of the flowers of Musa paradisiaca (high dose).

Experimental procedure:

Animals of all the groups will be fasted for 16-18hours before experimentation and fasting will be continued till the end of experimentation. However, the animals will be allowed to have free access to the water throughout the period of experimentation. A 12 hours light and 12 hours dark cycle is maintained with relative humidity of 45-55% and the animals will be maintained at an ambient temperature throughout period of experiment.

Before administration of vehicle /glibenclamide /extract, blood samples will be collected from the overnight fasted animals to determine the basal glucose level. Next the animals of respective groups will be administered with vehicle/glibenclamide /extracts and there after blood samples will be collected at 1, 2, 4, 6, 8, 12, and 24 hours intervals and analyzed for glucose concentration using GOD/POD method.

STATISTICAL ANALYSIS

All values will be expressed as mean ± SEM from 6 animals. Statistical difference in mean will be analyzed using one way ANOVA (analysis of variance) followed by Dunnett’s ‘t’test. p<0.05, will be considered as statistically significant.

7.3 Does the study require any investigation or intervention to be conducted on patients other humans or animals? If so, please describe briefly

Study requires investigation on rats. The effect of extracts of flower of Musaparadisiaca will be studied on various parameters as stated above in objectives of the study.

7.4 Has ethical clearance been obtained from your institute in case of 7.3?

YES:IAEC NO: 576/2002/IAEC/CPCSEA

ENCLOSURE-VI

List of References:

  1. Georg P, Ludvik B. Lipids and Diabetes. J Clin Basic Cardiol.2000; 3:159-62.
  2. Nyholm B, Porsen N, Juhl CB, Gravholt CH, Butler PC, Weeke J, Veldhuis JD, Pincus S, Schmitz O.Assessment of insulin secretion in relatives of patients with type 2 (non-insulin dependent) diabetes mellitus: Evidnce ofearly cell dysfunction. Metabolism. 2000; 49:896-905.
  3. Gandjbakhch I, Leprince P, D’Alessandro C, Ouaattara A, Bonnet N, Varvous s, Pavie A. Coronory artery bypass graft surgery in patients with diabetes. Bull AcadNatl Med. 2005; 72:69-76.
  4. John E Gerich, genetic basics of type-II diabetic mellitus impaired insulin secretion versus impaired insulin sensitivity, Endocrine review, 1998 (4): 491-03.
  5. Piedrola G, Novo E, Escober F, Garica-Robles R. white blood cell count and insulin resistance in patients with coronary artery disease. Ann Endocrinol. 2001; 62:7-10.
  6. Yaryura-Tobias JA, Pinto A, Neziroglu F. Anorexia nervosa, diabetes mellitus, brain atrophy, and fatty liver. Int J Etiol Disorders. 2001; 30:350-74.
  7. Rang HP, Dale MM. Ritter JM. The endocrine system pharmacology. In: Pharmacology. LongmanGroup Ltd, UK1991; P.504-8.
  8. Goodman and Gillman’s, the pharmacological basics of therapeutics. 10th ed. New York: Mc Graw Hill; 2001.
  9. KM Nadkarni, AK Nadkarnni, Indian Materia Medica, Vol-1. Bombay Popular Prakashan. 2000:1.
  10. Jain SR, Sharma SN. Hypoglycemic effect of Musa Sapientum L. flowers. Planta Medica 1967; 4:439-42.
  11. Jain SR. Hypoglycemic Principle in Musa Sapientum L. its Isolation. Planta Medica 1968; 5:43-47.
  12. Gadgoli C, Jolly CI. Starch from Musa Paradisiaca. Indian Journal of Pharmaceutical Science 1990; 52 (1):36-38.
  13. Ojewole, J.A., Adewunmi, C.O. Hypoglycemic effect of methalonic extract of Musa Paradisiaca green fruits in normal and diabetic mice. Method Find Clin Pharmacol 2003; 25(6): 453.
  14. Costa MM. Effects of prolonged administration of Musa ParadisiacaL(banana), and antiulcerogenic substance, in rats. Phytotherapy Res 1997; 11:28-31.
  15. Rodriguez R. Estudio del effecto analgesico de la savia del pseudotallo de Musa Paradisiaca en ratas. Rev. Cubana Invest Biomed 1991; 137:54-58.
  16. Ledon RE. Estudio del pseudotallo del platano Musa Paradisiaca L como antiasmatico. Rev. CENIC 1988; 19:58-60.
  17. MR Perez Capote, G Martinez Sanchez, OS Leon Fernandez. In vitro antioxidant actions of Musa Paradisiaca leaf extract (acitan). Pharmacologyonline 2007; 1:539-44.
  18. Mangathayaru K, Umeshankar G, Muralitharan G, Cordairayen E, Vasantha J. Antimicrobial activity of some indigenous plants. Indian Journal of Pharmaceutical Science 2004; 66 (1): 123-25.
  19. Yakubu M.T., Akanji M.A., Oladiji. Male sexual dysfunction and method used in assessing medicinal plants with aphrodisiac potentials. Pharmacognosy Reviews 2007; 1 (1).
  20. Eren Akcicek, Semihotles, Dursun Esiyok. Cancer and its prevention by some horticultural and field crops in Turky. Asian Pacific Journal of Cancer Prevention 2005; Vol-6.
  21. Kailash P, Bharathi K, Srinivasan k. Evaluation of Musa (Paradisiaca Linn. Cultivar) “Puttubale” stem juice for anti lithiatic activity in albino rats. Ind Physiol and Pharmacol 1993; 7:337-41.
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  24. Chhanda Mallick, Rajkumar Maiti, Debidas Ghosh. Comparative study on antihyperlipidemic effects of separate and composite extract of seed of Eugenia Jambolana and root of Musa Paradisiaca in streptozotocin induced diabetic male rat. IJPT 2006; 5 (1): 7-33.
  25. Chhanda Mallick, Kausik Chatterjee, Mehuli GuhaBiswas, Debidas Ghosh. Antihyperglycemic effects of separate and composite extract of root of Musa Paradisiaca and leaf of Coccina Indica in streptozotocin induced diabetic male albino rat. African Journal of Traditional, Complementary and Alternatiye Medicines 2007; 4 (3):362-371.
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