Diabetes mellitus (DM) is a chronic condition caused by inherited and /or acquired deficiency in production of insulin by the pancreas or by the ineffectiveness of the insulin produced. Such a deficiency results in the increased concentration of the glucose in the blood which in turn damage many of the body’s systems in particular the blood vessel and the nerves. It is projected to become one of the world’s main disabler and killer within the next 25 yrs. Regions with greatest potential are Asia and Africa, where DM rates could rise to two-three folds1.
Pharmacological means (insulin and hypoglycemics) as well as non pharmacological means (diet and exercise) may be used in the management of DM. The obvious limitations of these management methods necessitate a search for help among the arsenal of herbs available to man1.
To treat diabetes a number of synthetic drugs are available , but unfortunately the risk of severe hypoglycemia associated with the use of these medications can complicate other associated conditions such as congestive heart failure , renal insufficiency or liver disease or the medications may not be tolerated because of gastrointestinal adverse effects. Food and herbs have a pivotal role in traditional eastern medicine. The WHO Expert Committee on diabetes recommended further evaluation of the folkloric methods of managing this disease, because of the high mortality and morbidity arising from its attendant complications and problems associated with the use of conventional ant diabetic agents.2
Several indigenous medicinal plants are employed in the traditional management of diabetes mellitus but there is need to conduct pharmacognostic and pharmacological studies to ascertain their therapeutic values. The medicinal plants might be a useful source for the management of diabetes and its complications, for development of new chemical entities or as a dietary adjunct to existing therapies.
Stem wood of Cedrus deodara is used by practioners for the treatment of diabetes3. However, no systematic study was carried out on the stem wood extracts of Cedrus deodara for anti diabetic activity. Hence present study is proposed to investigate the hypoglycemic activity in normal and anti diabetic activity in alloxan induced diabetic rats.
7.0
7.1
7.2 / 6.2REVIEW OF THE LITERATURE:
Following are some of the plants having antidiabetic activity.4
Sl NO. / Name of the Plant / Parts Used
1) / Acacia arabica / Seed powder.
2) / Aegle marmelose / Root bark.
3) / Aloe barbadensis / Whole part.
4) / Areca catechu / Seeds.
5) / Azadirachta indica / Whole part.
6) / Beta vulgaris / Root.
7) / Biophytum sensitivum / Leaves.
8) / Bombax ceiba / Leaves.
9) / Brassica juncea. / Seeds.
10) / Caesalpinia bonducella / Seeds.
11) / Capparis decidua / Fruits.
12) / Citrullus colocynthis / Fruits.
13) / Coccinia indica / Leaves.
14) / Eucalyptus globulus / Leaves.
15) / Ficus bengalenesis / Barks.
16) / Gymnema sylvestre / Leaves.
17) / Hibiscus rosasinesis / Whole part.
18) / Momordica cymbalaria / Fruits.
19) / Momordica cymbalaria / Fruits.
20) / Vinca rosea / Leaves.
PLANT: Indian tree
SCIENTIFIC NAME: Cedrus deodara
FAMILY: Pinaceae
SYNONYMS5:
English names : Deodar , Himalayan cedar ,True cedar, Indian cedar
Sanskrit names : Devdaru , Mastadaru , amara,amaradaru
Hindi names : Deadwar .Deodar
Tamil names : Tevataram , tevataru.
PLANT DECRIPTION :
Cedrus is a genus of pinacea with basically tropical and subtropical worldwide distribution .The genus is comprised of trees which are sometimes cultivated either for their usefulness to traditional cultures or for ornamental purposes .It is a species of cedar native to western Himalayas in eastern Afghanistan, northern Pakistan, north west and central India, southwestern Tibet and western Nepal occurring at 1500-3200m.In India it is common from Kashmir, especially krishnaganga, kishtwar and jhelum to garhwal.It is a large corniferous tree reaching up to 50m height; with a trunk upto 3m diameter.Flowers appear in September and October. The wood has a character aromatic odour.3
Leaf : Evergreen needles , dark green but may have some silvery bloom giving them a blue green color; 1-2 inches long , sharp pointed ; occur singly on new growth and than later on spur shoots; remaining on the tree for 3 to 6 years.6
Flower: Monoecious; male cones 2 to 3 inches long on the lower parts of crown; female cones erect, purplish, occur on upper portions of crown.6
Fruit: Upright cones, 3 to 4 inches long and 3 inches across; deciduous scales; initially green and purplish, then later turning a reddish brown when mature, usually resinous.6
Twig: Slender, with numerous short spur shoots, branches droop with age; buds are very small and round.6
Bark: Initially smooth and gray-brown, later developing short furrows with scaly ridge tops.6
CHEMICAL CONSTITUENTS:
1) Terpenoids : a new novel type of phenolic sesquiterpene, himasecolone , has been isolated in addition to isopimaric acid from the choloroform soluble fraction extract from Cedrus deodara.7
2) Isohimachalone : Isolation and structure elucidation of a new bicyclic sesquiterpene of a new carboskeleton from the essential wood of Cedrus deodara is reported .8
3) Deoadraone and atlantolone : Isolation and structure determination of a novel bisabolone based tetra hydro -∆- pyrone from the essential wood of Cedrus deodara is reported.9
4) Oxidohimachalene : Isolation and structure elucidation of Oxidohimachalene a minor constituent of the essential oil of Cedrus deodara is reported.10
5) Isocentadarol : The constitution of isocentadarol a sesquiterpenoid from the Cedrusdeodara is reported. 11
6)Deodarin : a new dihydroflavanol named deodarin has been isolated from the stem bark of Cedrus deodara .12
7) Neolignan : A new neolignan and other phenolic constituents from the Cedrusdeodara has been isolated .13
8) Deodardione and sesquiterpene diosphenol : Isolation and structure elucidation of a C11 monocarboxylic acid , apparently a new sesquitepene , and a sesquiterpene diosphenol from the essential oil of Cedrus deodara are described .14
Medicinal Uses :
Antidote; Astringent; Carminative; Diaphoretic; Diuretic; Skin; Tuberculosis.
The heartwood is carminative, diaphoretic and diuretic.[15]
It has been used as an antidote to snake bites[15, 16].
The bark is astringent. It has proved useful in the treatment of fevers, diarrhoea and dysentery[15, 16].
It is used in the treatment of fevers, flatulence, pulmonary and urinary disorders, rheumatism, piles, kidney stones, insomnia, diabetes etc[15, 17].
The plant yields a medicinal essential oil by distillation of the wood, it is used in the treatment of phthisis, bronchitis, blennorrhagia and skin eruptions[18, 19, 15].
In Ayurvedic medicine the leaves are used in the treatment of tuberculosis[17].
OBJECTIVE OF THE STUDY:
# Preparation of Plant extract andits Preliminary Phytochemical evaluation.
# Determination of acute oral toxicity as per OECD guidelines.
# To evaluate hypoglycemic activity of extracts of Cedrus deodara in normal rats.
# To evaluate antidiabetic activity of Cedrus deodara in alloxan induced diabetic
rats.
MATERIALS AND METHODS
SOURCE OF DATA :
The plant material will be collected from the authenticated source. Whole experiment is planned to generate data from the laboratories studies. Experiment will be performed as described in the standard bibliography, may be obtained from standard journals and text books available within the college or from other Pharmacy colleges or from libraries of National Institutes or through internet or libraries from industry.
METHODS OF COLLECTION OF THE DATA (including sampling procedure if any):
The whole study is divided into following phases;
Phase I: Collection of plant material.
The plant material will be collected from the authenticated source in the month of February and March.
Phase II: Preparations of extracts.
Dried powder material successively extracted with petroleum ether and alcohol in a soxhlet apparatus for 72 hours. The residue obtained after alcoholic extraction is dried and macerated with chloroform water for 72 hours. The marc is strained and filtrate obtained is evaporated and reduce temperature to dryness. Percetage yield of all the three extracts will be determined.
Phase III: Preliminary Phytochemical investigation.
Preliminary phytochemical investigation will be done as described by Khandelwal20.
Phase IV:
Acute oral toxicity
Female Swiss albino mice (18-22 g) are individually identified and allowed to acclimate to the laboratory conditions for 7 days before the start of the study. Only one mouse receives a dose at a particular time. First animal receives a dose of 175mg/kg, po. Animal is observed for 48 hours after injection, for any toxicity signs, survival or death. If the first animal died or appeared moribund, the second animal receives a lower dose (55mg/kg). The dose progression or reduction factor is 3.2 times of the previous dose.
If no mortality is observed in the first animal then the second animal receives a higher dose (550mg/kg). Dosing of the next animal is continued depending on the outcome of the previously dosed animal for a fixed time interval (48 hour). The test is stopped when one of the stopping criteria is observed:
- 5 reversals occur in any 6 consecutive animals tested.
- 3 consecutive animals died at one dose level.
Statistical Analysis
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 test p<0.05 will be considerable significant.
Phase V: Pharmacological evaluation.
Drugs: Rosiglitazone , Alloxan etc.
Animals: Male albino wistar rats of body weight 170-220 g will be selected for allthe experiments. Animals will be kept in our animal house at an ambient temperature of 250C and 45-55% relative humidity with a 12h dark: 12 hour light cycle. Animals will have free access to food (rat pellet) and water.
GROUPS:
Two sets of a groups of rats consisting of 8 animals in each is divided as follows:
Group I : Vehicle control.
Group II : Standard drug (Rosiglitazone).
Group III : Petroleum ether extracts of low dose of Cedrus deodara .
Group IV : Petroleum ether extracts of medium dose of Cedrus deodara.
Group V : Alcoholic extracts of low dose of Cedrus deodara.
Group VI : Alcoholic extracts of medium dose of Cedrus deodara.
Group VII : Aqueous extracts of low dose of Cedrus deodara.
Group VIII: Aqueous extracts of medium dose of Cedrus deodara .
Group IX : Normal untreated rats.
Models: ASSESSMENT OF ANTIDIABETIC ACTIVITY OF Cedrus deodara
Model 1.1: Assessment of hypoglycemic activity in normal healthy rats.21
Initial testing was carried with the doses of the extracts prepared in normal healthy male rats fasted over night. The animals were divided into 9 equal groups as mentioned above. Blood glucose levels will be estimated before and after 2, 4, 6 and 8 h of extract administration.Model 1.2: Assessment of activity of extract on glucose tolerance in normal healthy rats.
On the next day after the assessment of hypoglycemic activity fasting blood glucose (FBG) will be checked in overnight fasted rats. The rats of all the groups will receive glucose (3 g/kg) after 90 min of extract or drug administration. Blood samples will be collected by retro orbital puncture method under light ether anesthesia just prior to glucose administration (0h) and 1, 2 and 3 h after glucose loading.
Model 2 : Assessment of anti diabetic activity in alloxan induced rats22
Induction of diabetes:
A fresh solution of Alloxan (75 mg/kg b.w.) in water will be prepared and injected intraperitonealy to overnight fasted rats. FBG level will be estimated at the time of induction of diabetes and post-prandial glucose level (PPG) will be checked regularly up to stable hypoglycemia, usually 1 week after alloxan injection. Animals having glucose between 250 to 350 mg/dl will be considered as diabetic and will be selected for further experiment.
Model 2.1 : Assessment of Antidiabetic activity of Cedrus deodara.
Diabetic animals will be divided into 9 groups of 8-10 animals and they will be dosed with extracts of Cedrus deodaraas mentioned earlier for 7 days. Daily blood glucose levels, body weight and urine sugar will be recorded in these diabetic animals.
Model 2.2 : Assessment of activity of extract on glucose tolerance in sub and mild diabetic rats.
On the 8th day extracts of Cedrus deodararoots were subjected to glucose tolerance test in diabetic animals. The rats of all the groups will give glucose (3 g/kg) after 90 min of the extract and drug administration. Blood samples will be collected just prior to glucose administration (0h) and 1, 2, 3 h after glucose loading.
Statistical analysis
The values will be expressed as mean±SEM. The data will be analyzed by using one way ANOVA followed by Donnett’s. Statistical significance will be set at P≤ 0.05.
Parameters to be evaluated :
Blood glucose, body weight of animal and urine sugar level will be estimated.
7.3 Does the study require any investigations or interventions to be conducted
on patients or other humans or animals? If so please describe briefly.
Yes, study requires investigation on Albino rats.
7.4 Has Ethical Clearance been obtained from your institution in case of 7.5?
Yes ethical clearance has been obtained.(copy enclosed)
8.0 / REFERENCES :
1.Osadebe PO, okide GB, Akabogu IC. Study on anti-diabetic activities of crude methanolic extracts of loran thus micranthus (Linn.) sourced from five different host trees. Journal of Ethno pharmacology 2004;95:133-8.
2.Swami Sadashiv , The ayurvedic encyclopedia , Sri Satguru publication a division of Indian book centre , Delhi , India chapter 4 . Pg 72-3.
3. Nadkarni KM, Nadkarni AK, editors. Indian Materia Medica. Bombay; Popular Prakashan 1976.
4. Grover JK, Yadav S, Vats V. Medicinal plants of India with anti-diabetic potential. J of Ethnopharmacol 2002;81:81-100.
5. Purohit, Prajapati, Sharma, Kumar. A Hand book of Medicinal Plants;
6. 10th oct.2007)
7.Agarwal.P.K.Rastogi. Terpenoids from Cedrusdeodara J of Ethnopharmacol.1981;20(6):1319-21.
8. Shankarnarayan R. Krishanapppa S. and Sukh Dev, Studies in sesquiterpenes LII-isohimachalone a novel sesquiterpenoids from the wood of Cedrus deodara J of Ethnopharmacol.1977;33(8):885-6.
9. Shankarnarayan R. Krishanapppa S. ,Sukh Dev,and Bisaraya SC Studies in sesquiterpenes LII-Deodarone and atlantone new sesquiterpenoids from the wood of Cedrus deodara J of Ethnopharmacol.1977;33(10):1201-5
10.Shankarnarayan R. Krishanapppa S. and Sukh Dev Studies in sesquiterpenes LIV- Oxidohimachalene new sesquiterpenoids from the wood of Cedrus deodara J of Ethnopharmacol.1977;33(10):1207-10.
11. Adinarayana A. Seshadari Chemical investigation of the stem bark of Cedrus deodara Isolation of anew dihydroflavanol deodarin. J Ethanopharmacology1965;21(12):3727-30
12.Kulshrestha DK , Rastogi. The constitution of Isocentradol ,sesquiterpenoidal from Cedrus deodara J Ethanopharmacology 1976;15(4):557-8
13. Agarwal.P.K. Agarwal.S.K. Rastogi A new neolignan and other phenolic constituentsfrom Cedrus deodara J Ethanopharmacology 1980;19(6):1260-1.
14. Krishanappa S and Sukh dev .Studies in sesquiterpenes –LVIII Deodardione a sesquiterpene diosphenol and limonenecaeboxylic acid , a possible Norsesquiterpene – compounds from the wood of Cedrus deodara J Ethanopharmacology 1978;34(5):599-602.
15. Chopra R. N.Nayar .S.L & Chopra .I.C. glossary of Indian medicinal plants(Including the supplements) CSIR . New Delhi.
16. Medicinal plants of Nepal . Dep. of medicinal plants. Nepal 1993
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17. Chevallier A.The encyclopedia of medicinal plants Darling , Kindersley. London 1996 . ISBN 9-780751-303148.
18. Grieve A Modern Herbal .Penguin 1984 ISBN 0-14-046-440-9.
19. Gupta B.L. Forest flora of Chakrata , Dehradun and Saharanpur FRI Press 1945.
20. Khandelwal KR.Practical Pharmacognosy-techniques and experiments. Pune, India: Nirali Prakashan; 1996
21. Kesari AN, Gupta RK, Singh SK, Diwakar S, Watal G. Hypoglycemic and antihyperglycemic activity of Aegal marmelos seed extracts in normal and Diabetic rats. J of Ethnopharmacol 2006;107:374-9.
22. Vogel HG editors. Drug Discovery and Evaluation, Pharmacological Assay, Second Edition; Germany; Springer-Verlag Berlin Heidelberg 2002
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