II.3.4.2. Evaluation of the antihyperglycemic activity in
normoglycemic rats
Test of antihyperglycemia provoked by oral route in normal
rats (oral glucose tolerance test): To evaluate the capacity of the organism to
manage glucose, 3 groups of 5 rats were constituted and treated as shown in
table IX below:
Table IX: Repartition of animals for the
antihyperglycemic test
|
Groups
|
Treatment
|
|
Negative Control
|
Distilled water (control)
|
|
Positive Control
|
2g/kg body weight of glucose
|
|
FOHF 300mg/kg BW
|
2g/kg BW of glucose + 300mg/kg body weight of hydroethanolic
fruit extract
|
|
FOHT 300mg/kg BW
|
2g/kg BW of glucose + 300mg/kg body weight of hydroethanolic
twigs extract
|
All the rats being at fasted state throughout the experiment,
the fasting blood glucose test of the various rats was done before
administration of our extracts (0h). The extracts were administrated orally by
intra-gastric route using an oesophageal probe, 30mins after, 2g/kg body weight
of glucose (test 1 and 2) was administration followed by successive glucose
test measurement determined at instants of 0.5h, 1h, 2h. Glucose measurement
was done by incision of the tail, a drop of blood placed on the glucose strip
and the glucose meter reading registered.
II.3.5. Evaluation of the
modulation effects of hydroethanolic fruits and twigs on some biomarkers of
diabetes type 2 on rat subjected to high fructose-high cholesterol diet
Table X: Slightly
modify food composition as proposed by Dhandapani (2007)
|
Composition
|
Control (%)
|
Atherogenic diet + fructose
|
|
Proteins (powder milk)
|
22
|
20
|
|
Carbohydrates (peel corn flour and wheat flour
(3:2)
|
55
|
50
|
|
Fructose
|
0
|
5
|
|
Lipids (hydrogenated fats and margarine)
|
15
|
17
|
|
Mineral salts (bone flour and NaCl (3:1)
|
4
|
4
|
|
Vitamins
|
1
|
1
|
|
Fibres (Cellulose)
|
3
|
3
|
For the preventive treatment, 4
groups of 5 rats were constituted as follows;
Table XI:
Repartition of animals for the preventive treatment with extracts
|
Groups
|
Treatment
|
|
Negative control
|
Non atherogenic diet and distilled water
|
|
Positive control
|
Atheregenic diet and gavages with 2ml of fructose(10%) + 2ml of
cholesterol (1.5%) 3 times a week
|
|
FOHF 300mg/Kg BW
|
Same as positive control and gavages with 300mg of FOHF/Kg of
BW everyday
|
|
FOHT 300mg/Kg BW
|
Same as positive control and gavages with 300mg of FOHT/Kg of
BW everyday
|
Experimental procedure
Male Wistar rats (175-225g) were divided into four
groups of rats. Food and water was provided ad libitum. The body
weights of rats were measured after 12 hours fasting period before
experimentation begins. The control group received just distilled water while
the three other groups received fructose and cholesterol by the intra-gastric
route. Test groups received FOHT and FOHF. The positive control received no
treatment. Fructose was administrated one hour before the administration of the
various extracts. The weights were taken again on day 7 and day 14
(Dhandapani et al., 2007, Idowu et al.,
2010).
At the end of the experimental period (14 days), animals were
kept 12 hours without food and water and their blood glucose taken. Fasting
rats were sacrificed under light anaesthesia with ether vapour. Blood collected
in EDTA tubes from the jugular artery was centrifuged for 10min at 3400 rpm and
the supernatant constituting the plasma was collected into dry eppendorf tubes
and stored at -20°C for some biochemical analysis. Tissue homogenate was
done by collecting the heart free from fat, and blood by rinsing in a solution
of 0.9% (w/v) NaCl. After anatomical, macroscopical, and comparative
observation, 1g of the organ was grinded in a mortar and homogenized in 10 ml
of 0.9% (w/v) NaCl. The resulting homogenate was put in a centrifuge tube,
allowed to sediment for about 1 hour before centrifuging at 3400 rpm for 10
minutes. The resulting supernatant which constitutes the homogenate was kept in
eppendorf tubes and conserve -20°C for later use in biochemical assay.
| 
