RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

KARNATAKA, BANGALORE

M. PHARM SYNOPSIS

YEAR OF ADMISSION-NOVEMBERS 2011

TITLE OF THE SYNOPSIS

“Antidiabetic activities of Phragmites karka (Retz.) in alloxan induced diabetic rats.”

BY

Mr. BHARATH. C L

DEPARTMENT OF PHARMACOLOGY

UNDER THE GUIDANCE OF

Mrs. Sarita. Kotagiri M.Pharm.

Asst. Professor

DEPARTMENT OF PHARMACOLOGY

INSTITUTION

GAUTHAM COLLEGE OF PHARMACY

R. T. NAGAR, BANGALORE-32

KARNATAKA

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

KARNATAKA, BANGALORE

ANNEXURE - II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / Name of the Candidate
And Address / Mr. BHARATH.C L
PERMANENT ADDRESS
S/o,K.Chandrappa
# 96, Krishna complex main
road , devanahalli
Banglore-562110
PRESENT ADDRESS
Gautham College of Pharmacy
Bhuvaneswari nagar,
R.T.Nagar Post,
Bangalore- 32.
2. / Name of the Institution / Gautham College of Pharmacy
Bhuvaneswari nagar,
R.T.Nagar Post,
Bangalore- 32.
3. / Course of Study and Subject / Master of Pharmacy in Pharmacology.
4. / Date of Admission / 29 - 9 - 2011
5.  Title of the Topic:
“Antidiabetic activities of Phragmites karka (Retz.) in alloxan induced diabetic rats.”

6.0 BRIEF REVIEW OF THE INTENDED WORK:

6.1 INTRODUCTION:

Diabetes mellitus is a chronic disease that occurs either when the pancreas does not produce enough insulin or when the body cannot effectively use the insulin it produces. Insulin is a hormone that regulates blood sugar. Defective insulin secretion is the major cause for chronic hyperglycemia resulting in impaired function or serious damage to many of the body’s systems, like eyes, kidneys, nerves, heart and blood vessels (1, 2). The common signs and symptoms are excessive thirst and urination, weight loss or gain, fatigue, and influenza–like symptoms. Early diabetes symptoms can be very mild and often even unnoticeable. Diabetes mellitus is one of the common metabolic disorders 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, 4).

Diabetes mellitus is a group of syndromes characterized by hyperglycemia, altered metabolism of lipids, carbohydrates and proteins and an increased risk of complications from vascular diseases. Most patients can be classified clinically as having either Type 1 diabetes mellitus (IDDM). It is an auto immune type I the main cause of this beta cell loss is a T-cell mediated autoimmune attack. It is also characterized by loss of the insulin-producing beta cells of the islets of Langerhans in the pancreas, leading to a deficiency of insulin (5). Type2 diabetes mellitus (NIDDM) is characterized differently due to insulin resistance or reduced insulin sensitivity, combined with reduced insulin secretion. Variants in 11 genes significantly associated with the risk of Type 2 diabetes of these 8 genes are responsible for impaired beta-cell function (6).

Health is defined as soundless of physical, mental or moral condition, especially freedom from bodily pain or disease, but true health is more than that. It includes the joy of living, the power and ability to lead a satisfying and purposeful life.

The World Health Organization has estimated that perhaps 80% of earths 6 billion inhabitance rely upon traditional medicine for their primary health care needs, and a major part of this therapy involves the use of plant extracts or their active principles.

6.2 NEED FOR THE STUDY:

“The aim of treating antidiabetic is to achieve present well being and future health”.

Diabetes Mellitus (DM) currently is a major health problem for the people of the world and is a chronic metabolic disorder/ syndrome resulting from a variable interaction of hereditary and environmental factors and is characterized by abnormal insulin secretion or insulin receptor or post receptor events affecting metabolism involving .carbohydrates, proteins & fats in addition to damaging liver, kidney and β-cells of pancreas (7)

Type-I (IDDM) diabetes is an auto immune disease caused by the destruction of pancreatic islets cells (8) whereas type-II (NIDDM) is due to the combination of Insulin resistance and a loss of secretary function by pancreatic β-cells. It is characterized by elevation of both fasting and post prandial blood sugar levels. Chronic hyperglycemia during diabetes causes glycation of body that in turn leads to secondary complications affecting eyes, kidneys, nerves and arteries (9)

In modern medicine, no satisfactory effective therapy is still available to cure the diabetes mellitus. Though insulin therapy is also used for the management of diabetes mellitus, but there are several drawbacks like insulin resistance, anorexia nervosa, brain atrophy and fatty liver after chronic treatment (10,11). Besides the use of insulin for the treatment of insulin dependent diabetes mellitus (IDDM), other approaches for the control of hyperglycemia include the use of amylin analogues. Sulphonylureas, the most widely used class of drugs act by closure of ATP dependent channel. Metformin, a Biguanide oral antidiabetic limits intestinal glucose absorption. These drugs have certain effects like causing hypoglycemia at higher doses, liver problems, lactic acidosis and diarrhea.

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 seven million new diabetic cases in the world (3).

There are reports that Phragmites Karka (Retz.) is used traditionally as antidiabetic plant, but there is no scientific antidiabetic activity had been carried out by this plant in rats. So, present study selected Phragmites Karka (Retz.) leaves for evaluation of antidiabetic activity by using plant extracts rats.

6.3 REVIEW OF LITERATURE(13):

Botanical name: Phragmites karka (Retz.)

Vernacular Name

Kannada: Hulugila hullu

Sanskrit: Dhamana, Nala

Synonym: Arundo karka, Arundo roxburgi, Phragmites roxburgi

Family: Poaceae

Distribution: India, Australia and Europe

Plant description: (12)

Aperennial reedwith long rhizomes and robust, erect culms to 3 m. The leaves are 15-30 cm long and nearly 2.5 cm broad; inflorescence is a large plume-like panicle with capillary branches and small, slender spikelets. It is leafy up the panicle. Phragmites can be easily distinguished from Arundo and Neyraudia by the silky beards at the bases of the lowest panicle branches, which are absent in the other two. InNew Guineathe reed occurs from near sea-level to at least 2000 m. It thrives in a rainfall regime from 200 to 5000 mm in swamps (India). It grows in standing water and is therefore tolerant of flooding. It usually grows in clay soils ranging from strongly acid (pH 4.5) to slightly alkaline (pH 7.5).

Traditional uses:

In Ayurvedic medicine system of medicine, the plant is used for different ailments such as diuretic, diabetic and anti-emetic (14).

6.4 OBJECTIVES OF THE STUDY:

Antidiabetic Activity:

The main objective of the proposed work is to induce diabetes by using alloxan in rats and screening of Phragmites karka (Retz.) leaf extract for its antidiabetic activites. The whole study is divided into two phases.

Phase I:

·  Collection and authentication of dried powdered leaf material subjected to successive extraction by using petroleumether, methanol and water by using soxhlet apparatus.

·  To investigate preliminary phytochemical constituents present in the extracts.

·  Determination of LD50 value and dose selection for proposed activities (selection of an appropriate dose from LD50 value) those considered as respective doses.

Phase II:

To evaluate antidiabetic activity of leaves extracts of Phragmites karka (Retz.) link by using the experimental animal model like:

-  Single dose study in normal rats (15).

-  Oral Glucose Tolerance Test (OGTT) (16).

-  Alloxan induced diabetic rats (16, 19, 20).

Parameters to be studied

·  Blood glucose levels.

·  Lipid profile (HDL, LDL, VLDL, TG, and Total Cholesterol).

·  Body weight.

·  Hemoglobin (Hb).

·  Superoxide dismutase (SOD).

·  Thiobarbituric acid reactive substances (TBARS).

·  Glutathione.

·  Albumin.

·  Urea.

·  Total protein.

-  Dexamethasone induced insulin resistance model in rats (18).

Parameters to be studied (16,18, 20).

·  Blood glucose levels.

·  Lipid profile (HDL, LDL, VLDL, TG, and Total Cholesterol).

·  Body weight.

7.0 MATERIALS AND METHODS:

7.1  Source of Data:

The Source of data will be based on laboratory experiments on animals and also the data obtained from the literature.

1.  Standard Books:

·  Goodman and Gilmann’s: The Pharmacological basis Therapeutics.

·  Rang and Dale’s Pharmacology.

·  Bertram G Katzung Basic and Clinical Pharmacology.

·  Wealth of India.

·  Indian Medicinal Plants.

·  SN Yoganarasimhan. Medicinal Plants of India, vol-1.

2.  Internet sources:

·  Pub med.

·  Science Direct.

·  SCOPUS.

·  Helinet.

·  Ovid.

·  Open J gate.

·  DOAJ.

·  Chemical Abstracts.

·  CABI.

·  International Pharmaceutical Abstracts.

·  Google.

·  Wikipedia.

3.  Journal sources:

·  Indian Journal of Pharmacology.

·  Journal of Pharmacology and experimental Therapeutics.

·  Journal of Ethanopharmacology.

·  Phytochemistry.

·  American Journal of Pharmacology and Toxicology.

·  Fitoterapia.

7.2  Preparation of Extracts (20).

The powdered form of the leaves of Phragmites karka (Retz.) was taken and subjected to successive solvent extraction. The extraction was carried out with the following solvents in increasing order of polarity: Petroleumether, methanol, and water. The solvent was then distilled, evaporated and vacuum dried and stored in air tight container.

7.3 ANTIDIABETIC ACTIVITY:

A.  Experimental animals: Adult albino rats weighing approximately 150-200 g of either sex will be used for antidiabetic activities. And rats are used for the acute toxicological studies. The animals will be fed with standard diet and will be given water ad libitum.

B.  Acute toxicity study of crude extract: For this purpose female albino rats (150-200g) will be used. Fixed dose method (OECD guideline no.425) of CPCSEA will be adopted (21).

C.  Dose: A dose of Petroleum-ether (PEPK), methanol (MEPK) and aqueous (AEPK) extracts of Phragmites karka (Retz.) will be taken as per acute toxicity study (22).

D.  Grouping of animals: The animals are divided into six groups, each group consists of six rats and alloxan is administered once.

7.3  Grouping of Animals :

I.  Single dose study in normal rats

A.  In normal animals:

1.  Effect of extract and glibenclamide on normal blood glucose levels(15):

Albino wistar rats weighing 150-200 mg/kg were divided into five groups of six in each group. Animals were fasted overnight for 16 hrs prior to the experiment. The blood glucose levels were measured just prior to and 1, 2, 4, 8 and 12 hrs after drug administration.

Group-I: Distilled water will be supplied and serve as normal control.

Group-II: Standard drug (Glibenclamide) will be supplied at a dose of 5 mg/kg orally.

Group-III: A dose of PEPK (1/10th of LD50) will be supplied orally in distilled water.

Group-IV: A dose of MEPK (1/10th of LD50) will be supplied orally in distilled water.

Group-V: A dose of AEPK (1/10th of LD50) will be supplied orally in distilled

water.

Parameters: Blood glucose levels.

2.  Oral Glucose Tolerance Test(16):

The oral glucose tolerance test was performed in rats weighing 150-200g. The animals were fasted for 16 hr before the experiment but allowed free access to water. These Rats were divided into five groups, six in each group. Rats of all groups were loaded with glucose 2g/kg p.o 30 min after drug administration. Blood samples were collected from the tail vein prior to drug administration and at 30, 60, 90 and 120 min of glucose administration.

Group-I: Distilled water will be supplied and serve as control.

Group-II: Standard drug (Glibenclamide) will be supplied at a dose of 5 mg/kg orally.

Group-III: A dose of PEPK (1/10th of LD50) will be supplied orally in distilled water.

Group-IV: A dose of MEPK (1/10th of LD50) will be supplied orally in distilled water.

Group-V: A dose of AEPK (1/10th of LD50) will be supplied orally in distilled water.

Parameters: Blood glucose levels.

B.  In Alloxan induced diabetic animals17:

Female Wistar rats weighing 150-220g were used for this study. The animals were overnight fasted for 16h before the induction of diabetes. Diabetes was induced by a single dose of 120 mg/kg body weight of alloxan by intraperitoneal route. After a period of 2 days blood glucose levels were checked by snipping the tail of alloxan treated fasted rats.

The study was carried out for 21 days. Fasting blood glucose levels were measured before the administration of PEPK, MEPK and AEPK. It was recorded as 0 day. The doses of 1/10 of PEPK, MEPK and AEPK along with the standard (Glibenclamide) were given daily to the animals for 21 days. The blood glucose levels were checked on 0, 7, 14, and 21 day of the treatment period. Blood was collected from snipping of the rat tail.

Group-I: Distilled water will be supplied and serve as normal control (for this group alloxan is not administered).

Group-II: 5% Tween 80 will be supplied and serve as diabetic control.

Group-III: Standard drug (Glibenclamide) will be supplied at a dose of 5 mg/kg orally.

Group-IV: A dose of PEPK (1/10th of LD50) will be supplied orally in distilled water.

Group-V: A dose of MEPK (1/10th of LD50) will be supplied orally in distilled water.

Group-VI: A dose of AEPK (1/10th of LD50) will be supplied orally in distilled water.

Parameters to be study

1. Blood glucose levels.

2. Body weight.

3. Total protein.

4. Albumin.

5. Urea.

6. Hemoglobin.

7. Total Lipid profile.

8. SOD.

9. TBARS.

10. Glutathione.

C.  Dexamethasone induced insulin resistance model in rats (18).

The Dexamethasone induced insulin resistance model was performed in rats weighing 150-200 mg/kg were divided into five groups of six in each group. Animals were fasted overnight for 16 hrs prior to the experiment. Dexamethasone sodium phosphate induced 10 mg/kg S.C administered from day 0 to 10 once daily. The blood glucose levels were measured just prior to 0 and 11 day.

Animals were divided into 5 groups 6 in each. All the animals receive Dexamethasone except group-1. Group 2 was not administered any other drug and acted as the positive diabetic control (Dexamethasone control). Group III to Group V were administered dexamethasone 10 mg/kg S.C and doses of 1/10 of PEPK, MEPK, and AEPK.

Group-I: Distilled water will be supplied and serve as normal control

Group-II: A dose of 10 mg/kg of dexamethosone S.C.

Group-III: A dose of PEPK (1/10th of LD50) will be supplied orally in distilled water.