ESTABILSHMENT OF TYPE 2 DIABETIC MOUSE MODEL AND EVALUATION OF ABILITY TO CONTROL PLASMA GLUCOSE IN TYPE 2 DIABETIC MICE OF BENINCASA

HISPIDA

CONTENTS
I. Introduction II. Background III. Materials and Method IV. Results and Discussion V. Conclusion

I. Introduction:
1. Actual Situation:
In the world: 246 mil. patients (2006). In Vietnam: 3.5 mil. patients (2003). 3rd place in top ten serious diseases.

I. Introduction:
2. Purpose:
Successfully establishment of type 2 diabetic animal model. Initially evaluate the ability to control plasma glucose of Benincasa Hispida.

II. Background:
1. Diabetes Definition
2. Chemicals Inducing Diabetes Disease 3. Benincasa Hispida

II. Background:
1. Diabetes Definition
Definition: Is a condition in which the body either doesnt produce enough, or doesnt properly response to, insulin. Classification:
Type 1: Results from the body's failure to produce insulin. Type 2: Results from Insulin resistance (cells fail to use insulin properly)

II. Background:
2. Chemicals Inducing Diabetes Disease
Streptozotocin (C8H15N3O7 - 265.221 g/mol)
Powder form Soluble in water; ketone, alcohol. Effect on cells (insulin producing cells)

STZ

II. Background:
2. Chemicals Inducing Diabetes Disease
Nicotinamide (C6H6N2O - 122.12 g/mol)
Prevents the activities of toxic agents inside the body. Helps in improving the function of cells. Lower the need of insulin.

NA

II. Background:
3. Benincasa Hispida
Long time used in traditional medicine Folk remedies using leaves, seeds, rinds, etc.

Benincasa Hispida

II. Background:
3. Benincasa Hispida
Classification:
Kingdom: Phylum: Class: Order: Family: Genus: Species: Plantae Magnoliophyta Magnoliopsida Cucurbitales Cucurbitaceae Benincasa B. Hispida

II. Background:
3. Benincasa Hispida
Ingredients:
Protein: Carbohydrate: Calcium: Phosphorus: Iron: Carotin: Vitamin B1: Vitamin B2: Vitamin PP: Vitamin C: Other elements 0.4g 2.4g 19mg 12mg 0.3mg 0.01mg 0.01mg 0.02mg 0.3mg 16mg

III. Materials and Method:

1. Materials:
Animal model: Mus musculus var. albino mice
5-6 weeks old 20-30g

Instruments:
Autoclave Magnetic stirrer Spectrophotometer Laminar hood Incubator Microscopy

III. Materials and Method:

1. Materials:
Equipments:
Glucometer Test strips Syringe 1ml 25G, 3ml and 5ml Falcon tube 50 ml Cotton pads Beaker 50ml, 100ml, and 250ml Erlenmeyer Duran 250ml, 500ml, and 1000ml

Eppendorf 1.5ml Forceps Scissors Micropipette 20 - 200 l Micropipette 100 - 1000 l Micropipette type 200l and 1000l Absorbance tissue Metal tray

III. Materials and Method:

1. Materials:
Chemicals:
STZ dose 100mg/kg NA dose 120mg/kg and 240mg/kg D-glucose solution Insulin PBS Collagenase DNS Dithizone

Benincasa Hispida: 30-50g

III. Materials and Method:

2. Method:
Procedure:
Mus musculus var. Albino mouse - Inject STZ and NA with different doses: STZ (100mg/kg) with NA (120mg/kg) and STZ (100mg/kg) with NA (240mg/kg) Mouse treated with medication - Test sugar blood concentration - Test physiological target after 35 days - Dye pancreas Type 2 diabetic mouse model

- Fed with Benincasa Hispida (1g/day)

- Test plasma glucose at day 0, 5, 10, 15, 20, 25, 30 Evaluate the effect of Benincasa Hispida

III. Materials and Method:

2. Method:
Physiological Target Tests:
Blood Glucose Concentration Test Glucose Tolerance Test Insulin Tolerance Test Urea Glucose Concentration Test Insulin-Containing Serum Test Dye Pancreas with Dithizone

IV. Results and Discussion:

1. Establishment of type 2 diabetic mouse model:
Inject STZ and NA with different doses: STZ (100mg/kg) with NA (120mg/kg), and STZ (100mg/kg) with NA (240mg/kg)

IV. Results and Discussion:

1. Establishment of type 2 diabetic mouse model:
Mouses blood glucose concentration:
Group 1: STZ (100mg/kg) with NA (120mg/kg) 35 days Group 2: STZ (100mg/kg) with NA (240mg/kg) 35 days

IV. Results and Discussion:

1. Establishment of type 2 diabetic mouse model:
Mouses blood glucose concentration:

IV. Results and Discussion:

1. Establishment of type 2 diabetic mouse model:
Glucose Tolerance Test: After injection of D-Glucose (1.5mg/kg)

IV. Results and Discussion:

1. Establishment of type 2 diabetic mouse model:
Insulin Tolerance Test:

IV. Results and Discussion:

1. Establishment of type 2 diabetic mouse model:
Urea Blood Concentration:

IV. Results and Discussion:

1. Establishment of type 2 diabetic mouse model:
Urea Blood Concentration:

IV. Results and Discussion:

1. Establishment of type 2 diabetic mouse model:
Insulin-containing serum:
Choose the mice that attain following physiological targets: Plasma glucose concentration is higher than normal level (control level). Low insulin tolerance ability. Low sensitivity to exogenous insulin. High urea glucose concentration

IV. Results and Discussion:

1. Establishment of type 2 diabetic mouse model:
Insulin-containing serum:
The increase of plasma glucose is unrelated to insulin synthesized in the body

IV. Results and Discussion:

1. Establishment of type 2 diabetic mouse model:
Dye Pancreas by Dithizone

Picture: Islets of Langerhans dyed with Dithizone

IV. Results and Discussion:

1. Establishment of type 2 diabetic mouse model:
Conclusion:
The effect of STZ (100mg/kg) with NA (120mg/kg) on plasma glucose of mice is higher than STZ (100mg/kg) with NA (240mg/kg):
Higher plasma glucose concentration Lower glucose tolerance ability Lower sensitivity to exogenous insulin Higher urea glucose concentration

The dose STZ (100mg/kg) with NA (120mg/kg) had successfully constructed type 2 diabetic mouse model.

IV. Results and Discussion:

2. Evaluate the ability to control plasma glucose of Benincasa Hispida:
Use type 2 diabetic mice treated by STZ (100mg/kg) with NA (120mg/kg) The evaluation is carried out after 40 days of type 2 diabetes induction. Mice is devided into 3 groups:
Group 1: type 2 diabetic mice, fed with Benincasa Hispida with the dose of 1g/day Positive control group: treated with commercial drug, Metformine (20mg/kg) Negative control group: no treatment

IV. Results and Discussion:

2. Evaluate the ability to control plasma glucose of Benincasa Hispida:

IV. Results and Discussion:

2. Evaluate the ability to control plasma glucose of Benincasa Hispida:
The effects of Benincasa Hispida and Metformin on plasma glucose control ability are the same.

V. Conclusion:
The dose STZ (100mg/kg) with NA (120mg/kg) has better effect on plasma glucose concentration of Mus musculus var. Albino and doesnt cause the damage in islets of Langerhans. The effect of Benincasa Hispida on type 2 diabetes has been demostrated but there should be much more time to observe clearly the effect of Benincasa Hispida on controlling plasma glucose ability.

Thank You For Your Kind Listening!