Enclosure-i brief resume of the intended work: 1 general discussion




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ENCLOSURE-I

BRIEF RESUME OF THE INTENDED WORK:

6.1 GENERAL DISCUSSION:
Perennial herb Majorana hortensis (Sweet Marjoram) was grown in Egypt over 3000 years ago and believed to be north Africa, the middle east and part of India1 .An aromatic Mediterranean plant, (Orginam Majorana) hortensis, with small pale purple flowers and sweet-scented leaves, used for seasoning food and in salads: Leaves - raw or cooked2,3. Sweet marjoram is widely used as a flavouring for salad dressings, vegetables, legumes and oil.4

There are more than 125 million persons with Diabetes Mellitus in the world today, and by 2010, this number is expected to approach 220 million. Both type 1 and type 2 DM are increasing in frequency. The reason for the increase of type 1 DM is not known. The genetic basis for type 2 DM cannot change in such a short time; thus other contributing factors, including increasing age, obesity, sedentary lifestyle, and low birth weight, must account for this dramatic increase. In addition, type 2 DM is being diagnosed with remarkable frequency in preadolescents and adolescents. Up to 45% of newly diagnosed children and adolescents have type 2 DM.5

In India a number of plants are mentioned in ancient literature (Ayurveda) for the cure of diabetic conditions known as "madhumehe" and some of them have been experimentally evaluated and the active principles isolated19936. plants have been the major source of drug for the treatment of diabetes mellitus in Indian system of medicine Out of many, only a few have been evaluated as per modern system of medicine7.The use of medicinal plants for the treatment of diabetes mellitus dates back to the Ebers papyrus of about 1550BC (Vailey and day). Even after discovery and use of insulin and other modern hypoglycemic agents, the search for safer and more effective drugs of plant origin for the treatment of diabetes has continued.8

More than 1200 plants species have been found to exhibit antidiabetic properties .9 In Argentina, plants have long been used for the empirical treatment of diabetes. Infact, world ethno botanical, information about medicinal plants reports that almost 800 plants are used in the control of diabetes mellitus, although a few of them have been scientifically studied.10

Ethno botanical information indicates that more than 100 plants used has traditional remedies for the treatment of diabetes (Aigonkar 1979; Alarcon-aguilara et al., 2000). The hypoglycaemic activity of large number of these plants have been evaluated and confirmed in different animal models (Torres and Surez, 1980; Gupta et al., 1984 Karawya et al., 1984.,).

Even now, approximately 80% of the third world population is almost entirely dependent on traditional medicines. The ethno botanical details and the Anti diabetic potential of several plants are known at present11. Origanum majorana belongs to family Labiatae. Various species of Origanum are O.vulgare, O.sativum and O.majorana. O.vulgare has been proved to have anti-hyperglycaemic activity12, but the anti-diabetic studies on O.majorana has not yet been reported so we have chosen this topic for dissertation work.



6.2 NEED FOR THE STUDY :

The term diabetes is a complex and multifarious group of disorders characterized by hyperglycemia that has reached epidemic proportions in the present century. Infection is a leading cause of morbidity and mortality among the diabetic population. Diabetes is associated with vascular and renal dysfunction characterized by hypertension, dyslipidemia, micro albuminuria, macro albuminuria and glomerular mesangial expansion. Diabetes mellitus may present with characteristic symptoms such as polyphagia, polydypsia, polyuria, blurring of vision, and weight loss. In its severe forms, ketoacidosis or a non- ketonic hyperosmolar state may develop and lead to stupor, coma and in the absence of effective treatment to death.5

Research and drug development efforts over past several decades have provided valuable information that applies directly to improving outcomes in patients with Diabetes mellitus and have expanded the therapeutic armamentarium.13

The currently available oral anti hyperglycemic agents for clinical use have characteristic profile of side effects.14 Management of diabetes with agents devoid of any side effects is still a challenge to the medical system. This has led to an increase in the demand for natural products with Anti hyperglycemic activity having fewer side effects. In accordance with the recommendations of the WHO15 expert committee on diabetes mellitus, an investigation of Anti hyperglycemic agents of plant origin used in traditional medicine seems important. Many herbs and plant products have been screened for Anti hyperglycemic activity.16,17,18 The medicinal plant of Origanum species O.vlugare had been proved the antihyperglyceamic activity12, based on this evidence the plant O.majorana (Majorana hortensis) is selected in present study for its antidiabetic activity.



6.3 REVIEW OF THE LITERATURE :

Botanical Name: O. majorana

Family Name : Labiatae

Common Name: Majorana hortensis, sweet marjoram, annual marjorana, knotted marjoram19

A) Habitat.

Sweet marjoram is native to North Africa, Turkey and SW Asia. It has naturalized in the Mediterranean region of southern Europe.20 Sweet Marjoram grows about 12 inches tall as is mostly grown as an annual.21


B) Morphology Description (Habit).22

Flowers - small whitish, pink, purplish (or) pale pink in terminal clusters.

Seeds - minute, oval, dark brown.

Coralla - with short limb.

Parts used- whole plant.

Leaves - oblongovate.

C) Chemical constituents:22

The content of essential oil depends on soil, climate and season, but generally lies between 0.7% and 3.5%. The main aroma component is a bicyclic monoterpene alcohol, cis-sabinene hydrate (max. 40%); furthermore, α-terpinene, 4-terpineol, α-terpineol, terpinenyl-4-acetate and 1, 8-cineol are found in significant amounts. Phenolic compounds,



D) Ethnomedical uses:

Sweet marjoram is mainly used as a culinary herb, but is also medicinally valuable due to its stimulant and antispasmodic properties.23

The herb is antiseptic, antispasmodic, carminative, cholagogue, diaphoretic, diuretic, emmenagogue, expectorant, stimulant, stomachic and mildly tonic.24

It is taken internally in the treatment of bronchial complaints, tension headaches, insomnia, anxiety, minor digestive upsets and painful menstruation.25

Antioxidant properties of marjoram (Majorana hortensis) herb and extracts obtained with ethanol, n-hexane, and supercritical CO2 extraction are presented. Individual antioxidants, ursolic acid, carnosic acid, and carnosol, were quantified with high-performance liquid chromatography. The effects of different parameters (temperature and pressure) of high-

pressure extraction on the yield of carnosol were studied.26


Sweet marjoram is widely used as a flavoring for salad dressings, vegetables, legumes and oils. The leaves and flowers yield 0.3 - 0.4% essential oil by steam distillation. Called 'Oil of Sweet Marjoram', it is used as a food flavoring and in perfumery, soaps, hair products etc.24


6. 4. OBJECTIVES OF THE STUDY:


  1. Collection of Majorana hortensis plant.

  2. Extraction of whole plant of Majorana hortensis.

  3. Screening of Anti diabetic activity.

I. Alloxan­­- induced diabetes

ENCLOSURE-II

MATERIALS AND METHODS:


    1. Source Of Data:

1. Library, Bharathi College of Pharmacy.

2. E-library, Bharathi College of Pharmacy.


7.2 Experimental animals:

Adult albino rats weighing approximately 150-200 g will be used. The animals will be fed with standard feed and will be given water.


7.3 Plant material:

The whole plant of Majorana hortensis will be powdered coarsely and about 500 g of this powder will be extract (Soxhlet) with aqueous water.


7.4 Selection of dose:

The dose of 250 and 500 mg/kg will be used.28


7.5 Methods and Experimental design:

7.5.1 Grouping of animals: The animals are divided into six groups of five rats in each group.

1.Model for IIDM:



Group I : Normal control

Group II : Diabetes induced control

Group III: Diabetes induced + Standard drug treated.

(Standard drug : Glibenclamide 1mg/kg by oral route)



Group IV : Diabetes induced +Test drug treated Dose I.

Group V : Diabetes induced +Test drug treated Dose II.

. Group VI : Test drug treated Normal group.



Parameters to be evaluated:

    1. Blood glucose levels.

    2. HDL, LDL, Total cholesterol.

    3. Plasma triglycerides.

    4. Body weight.

7.5.2 Chemical induced diabetic model :27

Alloxan­­- induced diabetes:

Hyperglycemia and glucosuria after administration of alloxan has been described in several species, such as in dogs, in rabbits, in rats Goldner and Gomori and in other species .Guinea pigs have been found to be resistant. Dosage and treatment regimen have been elaborated for the most frequently used species. In most species a tri phasic time course is observed: an initial rise of glucose is followed by a decrease, probably due to depletion of islets from insulin, again followed by a sustained increase of blood glucose.


Evaluation:

Data will be analyzed using one way analysis of variance (ANOVA).


7.5.3 Experimental design:

Anti-diabetic activity:

S. No

Treatment

Dose

(mg/kg)

Route of administration

No. of.

Animals

1

Control (Distilled water)

_____

Oral

5

2

Alloxan treated

120

S.C.

5

3

Positive Control

Glibenclamide



1

Oral

5

4

Majorana hortensis

250

Oral

5

5

Majorana hortensis

500

Oral

5

6

Test Control

Majorana hortensis

1000

Oral

5


7.6. DOES THE STUDY REQUIRE ANY INVESTIGATION OR INTERVENTIONS TO

BE CONDUCTED ON PATIENTS OR OTHER HUMANS OR ANIMALS?


YES

Antidiabetic activity test: Adult albino rats weighing approximately 150-200 g will be used

to check antidiabetic activity of Majorana hortensis.

7.7. HAS ETHICAL CLEARANCE BEEN OBTAINED FROM YOUR INSTITUTION

IN CASE OF 7.6?

The animal study is cleared from ethical committee of the institution.

(Reg. no. 1135/a/07/CPCSEA, Dated: 13/02/2008) - Copy enclosed.




ENCLOSURE-III

REFERENCES:

  1. R Tainter, D.R. and A.T. Grennis, 1993. Spices and Seasonings: A Food Technology Hand Book, VCH Publishers, Inc. New York.

  2. Hedrick. U. P. Sturtevant's Edible Plants of the World. Dover Publications 1972 ISBN 0-486-20459-6.

  3. Grieve. A Modern Herbal. Penguin 1984 ISBN 0-14-046-440-9.

  4. www.Origanum majorana - Plants for a Future database report.html.

  5. Goodman & Gilman's. The Pharmacological Basis Of Therapeutics; 11th ed.ch16 2006; 1613.

  6. R Maiti,D. Jana, UK Das, D. Ghosh Antidiabetic effect of aqueous extract seed of Tamarindus indica in streptozotocin-induced diabetic rats. J Ethnopharmacology 2004; 92: 85-91.

  7. SB Sharma, A Nazir,KM Prabu,PS Murthy,G. Dev, Hypoglycemic and hypolidemic effect of ethanolic extract of seeds of Eugenia jambolana in alloxan-induced diabetic rabbits. J Ethnopharmcology. 2003; 85: 201-6.

  8. Mohammad Ali Esmaeili, Razieh Yazdanparast Hypoglycemic effect of Teucrium polium: studies with rat pancreatic islets. J Ethnopharmcology 2004; 95:27-30.

  9. M. Eddouks, Lemhadri, J.B. Michel. Caraway and caper: Potential anti-hyperglycaemic plants in diabetic rats. J Ethnopharmacology 2004; 94:143-8.

  10. Manuel J. Ayber, Alicia N. Sanchez Riera, Alfredo Grau, Sara S. Sanchez. Hypoglycemic effect of the water extract of Smallantus sonchifolious (yacon) leaves in normal and diabetic rats. J Ethnopharmacology 2001; 74:125-32.

  11. Nadkarni AN. Indian Materia Medica, vol. 1. Mumbai, India: Popular Book Depot; 1989. 

  12. Lemhadri A, Zeggwagh NA, Maghrani M, Jouad H, Eddouks M. Anti-hyperglycaemic activity of the aqueous extract of Origanum vulgare growing wild in Tafilalet region. J Ethnopharmacoly. 2004 Jun; 92(2-3):251-6.



  1. Triplitt L, Reasner A, Isley L. Diabetes mellitus. In: Dipiro T, Talbert L, Yee C, Matzke R, Wells G, Posey L, editors. Pharmacotherapy a patho physiologic approach. 6th ed. United States of America: Mc Graw Hill publications; 2005: 1334.

  2. Holman RR, Turner RC. Oral agents and insulin in the treatment of NIDDM. In: Pickup J, Willians G, editors. Textbook of Diabetes. Oxford: Blackwell; 1991

  3. The WHO Expert Committee on Diabetes mellitus. Technical Reports series 646. Geneva: World Health Organization; 1980.

  4. Bailey CJ, Day C. Traditional plants medicines as treatment for diabetes. Diabetes Care. 1989; 12:553-64.

  5. Mukherjee SK. Indigenous drugs in diabetes mellitus. J Diab Assoc India. 1981; XXI: 97-106. 

  6. Ajgaonkar SS. Herbal drugs in the treatment of diabetes a review. IDF Bulletin. 1979; 24:10-7.

  7. Dr.K.madhava chetty, K.sivaji and K.tulasi rao. Flowering plants of chittor: chittor. 2nd ed; 2008 pub: students offset printers; 278.

  8. www.Floridata Origanum majorana.htm.

  9. www.Sweet Marjoram Garden Guides.htm.

  10. The wealth of India; Raw materials L-M; vol4; reprinted -2003; NISCAIR; 227.

  11. R Chevallier. A. The Encyclopedia of Medicinal Plants Dorling Kindersley. London 1996 ISBN 9-780751-303148.

  12. www.Origanum majorana - Plants for a Future database report.htm.

  13. Bown. D. Encyclopaedia of Herbs and their Uses. Dorling Kindersley, London. 1995 ISBN 0-7513-020-31.

  14. E. Vági, E. Rapavi, M. Hadolin, K. Vásárhelyiné Perédi A. Balázs, A. Blázovics, and B. Simándi. Phenolic and Triterpenoid Antioxidants from Origanum majorana L. Herb and Extracts Obtained with Different Solvents J. Agric. Food Chem. 2005; 53 (1):17-21.

  15. H.Gerhard Vogel. Drug Discovery Evaluation: Pharamacological Assays. Germany.vol1, 3rd ed. Springer; 2007.



  1. Al-Howiriny, Tawfeq · Alsheikh, Abdulmalik · Alqasoumi, Saleh · Al-Yahya, Mohammed · ElTahir, Kamal · Rafatullah, Syed. Protective Effect of Origanum majorana L. 'Marjoram' on various models of gastric mucosal injury in rats. The American journal of Chinese medicine 2009; 37(3): 531-545.






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