Antidiabetic Activity Of Kyllinga Triceps Root Extracts Biology Essay

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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 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 3,4.

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 insulin5. Type 2 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 function6.

Kyllinga is a genus of flowering plants in the sedge family known commonly as spike sedges. They are native to tropical and warm temperature areas of the world especially tropical Africa. These sedges vary in morphology, growing to a height from 2 centimeters to a meter. These are closely related to cyperus species7.

The roots of the Kyllinga triceps is employed in the East Indies in diabetes, and as stomachic8. The decoction from the roots is used as refrigerant, demulcent and tonic. It is given to relive thirst in fever9.

6.2 NEED FOR THE STUDY :

"The aim of treating Diabetes 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 pancreas10.

Type-I (IDDM) diabetes is an auto immune disease caused by the destruction of pancreatic islets cells11. 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 arteries12.

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 13, 14. 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 diarrhea15.

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) and Germany (7.4 million). by the year 2025, there will be as many as seven million new diabetic cases in the world3.

It is apparent that due to the side effects of the currently used drugs, there is a need for a safe agent with minimal adverse effects, which can be taken for long durations. Recently, there has been increasing interest in the use of medicinal plants. The use of medicinal plants in modern medicine suffers from the fact that though hundreds of plants are used in the world to prevent or to cure diseases. Recently search for appropriate antihyperglycemic agent has been focused on plants used in traditional medicine because of leads provided by natural products that may be better treatment than currently used drugs.

There are reports that Kyllinga triceps is used traditionally antidiabetic plant, but there is no scientific antidiabetic activity has been carried out by this plant. So, present study selected roots of Kyllinga triceps plant for evaluation of antidiabetic activity by using streptozotocin induced diabetes rats.

6.3 REVIEW OF LITERATURE:

Botanical name: Kyllinga triceps (Rottb.)

Synonym: Cyperus triceps, K. tenuifolia

Family: Cyperaceae.

Distribution: Northwestern India, Gujarat, Rajasthan and South India,

Biological properties:

It is a small glabrous erect herb, with elongate creeping rhizomes. It is grown during the months of July to October16. The plant is used as "Nirvishaa" and "Apavisha" in Ayurvedic medicine. the plant is used as "Mustu" in Maharashtra as a folk medicine. The roots of the plant are used for febrifuge and antidermatosis17. The root is used for stimulating liver and to relieve pruritis 18, 19.

In Chhattisgarh this plant is called as Bendraphool and is used for the treatment in the wound healing purpose20.

6.4 OBJECTIVES OF THE STUDY:

The main objective of the proposed work is to induce diabetes by using streptozotocin, in rats and screening of Kyllinga triceps root extracts for its antidiabetic activity. The whole study is divided into two phases.

Phase I:

Collection and Authentication of plant material. Shade drying, powdering of root part of the plant and extraction of root part of the plant with different solvents ( Petroleum ether, methanol and aqueous) by using Soxhlet apparatus.

To investigate preliminary phytochemical constituents present in the extracts.

Determination of LD50 value and dose selection for anti-diabetic activity( selection of an appropriate dose from LD50 value) those considered as respective doses.

Phase II:

To evaluate anti-diabetic acivity of root extracts of Kyllinga triceps (Rottb) by using the experimental animal model like:

Streptozotocin induced diabetic rats.

It is also planned to evaluate the following parameters.

Blood glucose levels

Body weight

Total protein

Superoxide dismutase ( SOD )

Thiobarbituric acid reactive substances ( TBARS )

Glutathione

Albumin

Urea

Hemoglobin ( Hb )

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

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.

Standard Books: Goodman and Gilmann's: The Pharmacological basis of Therapeutics.

Internet source:

Google.

Pubmed.

Science direct

Wikipedia

SCOUPS

Helinet

Journal sources:

Indian Journal of Pharmacology

Journal of Pharmacology and experimental Therapeutics

Journal of Ethanopharmacology

Phytochemistry

7.2 Preparation of Extract21:

About 250 gm of powdered shade dried roots of Kyllinga triceps are subjected to successive soxhlet extractions by using petroleum ether, methanol, and water. The prepared extracts are concentrated to lesser volume under reduced pressure and 2w evaporate to dryness. The prepared extract is suspended in distilled water containing 2% v/v Tween 80 (Suspending agent).

7.3 Antidiabetic activity:

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

Plant material: Naturally available plant Kyllinga triceps will be collected, identified and extracted with petroleum ether, alcohol, and water.

Acute toxicity study of crude extract23: For this purpose female albino mice (20-25g) will be used. Fixed dose method (OECD guideline no. 420) of CPCSEA will be adopted.

Dose: A dose of petroleum ether extract(PEKT), methanolic extract(MEKT) and aqueous extract of Kyllinga triceps (AEKT) will be taken as per the work24.

Grouping of animals: The animals are divided into 5 groups of six rats in each group and the treatment given once.

7.4 Grouping of animals25:

EFFECT OF EXTRACTS:

In normal animals:

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

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

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

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

In Streptozotocin induced diabetic animals:

Animals will receive streptozotocin 60 mg/kg via I.P/ I.V route.

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

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

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

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

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

Glucose Tolerance Test22:

Overnight fasted rats were divided into 4 groups

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

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

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

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

7.5 Collection of Blood and Analysis Procedure:

Blood samples will be collected by Retro orbital sinus. The blood glucose level will be measured by Glucose oxidase / Peroxidase method. Statistical analysis will be done by ANOVA.

Blood glucose levels, Total protein, Superoxide dismutase ( SOD ), Thiobarbituric acid reactive substances ( TBARS ), Glutathione, Albumin, Urea, and Hemoglobin ( Hb) and Lipid profile(HDL, LDL, VLDL, TG, Total Cholesterol).

7.6 Histopathology;

The isolated pancreas are subjected to histopathological studies of α and β cells.

7.7 EXPERIMENTAL DESIGN:

7.7.1 EFFECT OF EXTRACTS:

In normal animals:

Sl.

No

Treatment

Dose

mg/kg

ROA

Duration of measurement

No. of animals

Parameters

1

Control

------

Oral

1,2,4,8,12,24 hrs

06

Blood glucose levels

2

PEKT*

1/10th of LD50

Oral

1,2,4,8,12,24

Hrs

06

3

MEKT**

1/10th of LD50

Oral

1,2,4,8,12,24

Hrs

06

4

AEKT***

1/10th of LD50

Oral

1,2,4,8,12,24

Hrs

06

*PEKT- Petroleum ether extract of Kyllinga triceps

**MEKT- Methanolic extract of Kyllinga triceps

***AEKT- Aqueous extract of Kyllinga triceps

B. In Streptozotocin induced diabetic animals:

Sl.

No

Treatment

Dose

mg/kg

ROA

Duration of measurement

No. of animals

Parameters

1

Control

------

Oral

1,7,14,21 days

06

1.Blood glucose levels

2. Body weight

3.Total protein

4.SOD

5.TBARS

6.Glutathione 7.Albumin

8. Urea

9. Hemoglobin

10.Total Lipid profile

2

Glibenclamide

5 mg/kg

Oral

1,7,14,21 days

06

3

PEKT*

1/10th of LD50

Oral

1,7,14,21 days

06

4

MEKT**

1/10th of LD50

Oral

1,7,14,21 days

06

5

AEKT***

1/10th of LD50

Oral

1,7,14,21 days

06

*PEKT- Petroleum ether extract of Kyllinga triceps

**MEKT- Methanolic extract of Kyllinga triceps

***AEKT- Aqueous extract of Kyllinga triceps

Glucose tolerance test:

Sl.

No

Treatment

Dose

mg/kg

ROA

Duration of measurement

No. of animals

Parameters

1

Control

------

Oral

30, 90, 150 min

06

Blood glucose levels

2

PEKT*

1/10th of LD50

Oral

30, 90, 150 min

06

3

MEKT**

1/10th of LD50

Oral

30, 90, 150 min

06

4

AEKT***

1/10th of LD50

Oral

30, 90, 150 min

06

*PEKT- Petroleum ether extract of Kyllinga triceps

**MEKT- Methanolic extract of Kyllinga triceps

***AEKT- Aqueous extract of Kyllinga triceps

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