PHARMACOLOGY PRACTICUM REPORT

Experiment 6

ANTIDIABETIC

Supporting lecturer: apt. Reza Alrayan, M. S Farm

Compiled by:

Group 3

1. Nanda Aditya (10123106)

S1 PHARMACY STUDY PROGRAM

FACULTY OF PHARMACY

INSTITUT ILMU KESEHATAN BHAKTI WIYATA KEDIRI

2023
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EXPERIMENT 6

ANTIDIABETIC

I. AIM TO

Determine the effect of drugs on reducing glucosec levels in the blood of

experimental animals.

II. TOOLS, MATERIAL, AND TEST ANIMALS

a. Tools
- Analitycal balance
- Injection syringe 1 ml
- Glucose meter
- Glucose test strips
- Sonde oral
- Cage mice
- Permanent marker
- Stopwatch
- Steril gloves
- Tissue/cotton
- Medical scissors
b. Materials
- Alcohol 70%
- Sterile alcohol
- Dextrose
- CMC Na suspension
- Glibenclamide 5 mg
- Metformin 500 mg
- Thick extract
c. Test animals
- Mice (Mus Musculus)
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III. CALCULATION

a. Mice Number 1 (Weight 26 g) => Glibenclamide

- Conversion dose: 0,0026 x 5 mg = 0,013 mg
- Mice dose: 26 g/20 g x 0,013 mg = 0,0169 mg
- Volume giving: 0,0169 mg/5 mg x 50 ml = 0,169ml=0,2ml

b. Mice Number 6 (Weight 30 g) => CMC Na = 1 ml

c. Mice Number 4 (Weight 24 g) => Metformine

- Conversion dose: 0,0026 x 500 mg = 1,3 mg
- Mice dose: 24 g/20 g x 1,3 mg = 1,56 mg
- Volume giving: 1,56 mg/500 mg x 50 ml = 0,156ml=0,2ml
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IV. METHOD

ANTIDIABETIC

Take 2 mice then weight the mice by lifting their tails and
placing them on the scales

Each Given Dextrose PO 1 ml

To increase blood sugar

Mice 1 (26 g) Mice 6 (30 g)

Glibenclamide (0,2 ml) CMC Na (1 ml)

1. Hold the mice by pinching the bend
using the tumb and index finger, and
the tail between the ring and little
fingers
2. When, holding the animal upside
down, make sure the head is looking
up or parallel to the body and the
mouth is slightly open
3. Insert an oral probe filled with 1 ml
dextrose close to the roof of the
mouse’s mouth
4. Then slide it down the esophagus and
force the solution out of the oral
needle
5. Then wait 30 minutes and check the
mice’s blood sugar
6. Then do steps 1-4 again with an oral
probe containing glibenclamide 0,2 ml
7. Then observe and wait 30 minutes to
check your blood sugar again
1. Hold the mice by pinching the bend
using the tumb and index finger, and the
tail between the ring and little fingers
2. When, holding the animal upside down,
make sure the head is looking up or
parallel to the body and the mouth is
slightly open
3. Insert an oral probe filled with 1 ml
dextrose close to the roof of the mouse’s
mouth
4. Then slide it down the esophagus and
force the solution out of the oral needle
5. Then wait 30 minutes and check the
mice’s blood sugar
6. Then do steps 1-4 again with an oral
probe containing 1 ml CMC Na
7. Then observe and wait 30 minutes to
check your blood sugar again
(This section is not given CMC Na and
lood sugar is not checked)
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V. RESULT

a. Table of abilities of each individual

TYPES OF NAME
ABILITIES Fatim Mira Miranda Dzaky Achsin Maharani Nanda
Handing Animal 95% 90% 90% 95% 95% 95% 50%
Oral 85% 75% - 90% 85% 80% -
Subcutaneous 50% - 35% - 95% 85% -
Intraperitoneal 80% - - - - - -
Intramuscular - - - - - - -
Intravenous - - - - - - -

Information: Type of ability is present with grades 1-100%

b. Weighing table of mice weights

Weight of Mice (grams)

No. Mice Week 1 Week 2 Week 3 Week 4 Week 5 Information
1 18 22 28 29 26
2 18 22 25 27 - -
3 22 26 31 33 29
4 18 22 26 29 27
5 14 19 24 26 26 =
6 21 23 30 31 30
7 24 28 31 32 -
8 15 21 25 27 26
9 15 20 24 - -
10 22 25 30 - -
Average 18,7 g 22,8 g 27,4 g 29,25 g 23 g

Information: Dead Mice

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c. Observations

No Animal Test Weight Treatment Initial Sugar level

sugar level after 30’
(mg/dL) (mg/dL)
1 Mice 1 26 Glibenclamide 183 89
2 Mice 8 30 CMC Na - -
3 Mice Group 4 24 Metformine 143 165

VI. DISCUSSION

Performance development, in group 3 there was progress in its members, where

the members were able to hold mice correctly but there were several members who
held them incorrectly and in this practicum they administered oral medication using
a probe well, because there was progress from last week which was proven with no
dead menvit after treatment. This is because in this practicum the drug
administration method used was oral drug administration, so that group 3 members
had the opportunity to practice holding mice correctly and administering the drug
orally better. There are some members who still do not have the courage to give oral
treatment. Group 3 members are also increasingly responsive by showing initiative
to immediately carry out existing tasks without having to wait for orders to carry out
tasks.

Development of mice for one week. In observations made on the body weight of
the mice, it was found that the body weight of all mice decreased. This is thought
to be due to the reduced frequency of visiting the mice in the cage, and the condition
of the mice being poorly looked after. Apart from that, there were several mice that
died, the cause of the last mouse that died was unknown because the mouse suddenly
disappeared and there was no trace of death. All the mice experienced weight loss,
this was because when they were given food, they always didn't run out and a lot of
the food turned into powder, since some of the mice died, the mice's
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appetite decreased. So the mice become malnourished which causes their body
weight to decrease.

In this practicum, the antidiabetic effect was tested on experimental animals,

namely mice. Diabetes mellitus, or what is often called diabetes, is a condition
characterized by high blood sugar levels in the body. High blood sugar occurs when
the body cannot produce or use insulin properly. Insulin is a hormone produced by
the pancreas and plays a role in regulating blood sugar levels. Diabetes can also
occur when the body is no longer able to take sugar (glucose) into cells and use it
as energy. This condition ultimately results in a buildup of extra sugar in the body's
bloodstream.

Treatment of experimental animals. This time Table 3 observed mice that were
given dextrosal orally to increase blood sugar levels and observed the effect after
being given the antidiabetic drug, namely Glibenclamide. Glibenclamide works by
stimulating the pancreas to increase the production and use of the hormone insulin
by the body. This hormone is responsible for entering blood sugar into body cells,
so that blood sugar levels can decrease. The first thing to do is weigh the mice to
find out their weight which will then be used to find the dose of drug and volume
that will be given to the mice orally. The mice used are mouse number 6 with a
weight of 30 grams which will be given Glibenclamide and number 1 with a weight
of 26 grams for negative control using CMC Na. Next, both mice were given 1 ml
of dextrosal orally to increase blood sugar levels and then left for 30 minutes so that
the dextrosal could work properly. After 30 minutes, mouse number 6 had a small
cut at the tip of its tail to obtain a blood sample from the mouse which would be
used to observe its blood sugar levels. After observing, it was found that the blood
sugar level of mouse number 6 was 188 mg/dl. This data shows that the sugar level
has exceeded the normal limit in mice according to the limits in the literature for
normal blood sugar levels in male mice, namely 62.8-176.0 mg/dl. etc. Due to
limited equipment for measuring blood sugar levels, the blood sugar level of mouse
number 1 was not measured, therefore the results from group 4 will be used as a
comparison.
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After measuring blood sugar levels, mouse number 8 was immediately given an
antidiabetic drug, namely Glibenclamide. The drug was given orally and then the
mice were left for 30 minutes, so that the drug could react optimally. After 30
minutes blood samples were taken again from the tip of the mice's tails and
measured to determine the blood sugar levels in the mice. The results of measuring
blood sugar levels in mice after Glibenclamide reacted were 89 mg/dl. This shows
a decrease in blood sugar levels in the mice blood samples, which indicates that the
experiment carried out was successful.

Comparison with Group 4. The results obtained from group 4 which was used as
a comparison showed opposite data from the experiment carried out by group 3.
Group 4 used Metformin as the antidiabetic drug used for this experiment. The
blood sugar level data obtained in mice before being given Metformin was 143
mg/dl, then the blood sugar level data obtained after being given Metformin was
165 mg/dl. From this data, it was found that blood sugar levels increased in mice
after being given antidiabetic drugs in the form of Metformin. This may occur due
to procedural errors or insufficient medication dosage and several other factors.

The obstacle encountered in this practicum, was in taking blood samples from
mice. The procedure for taking blood samples was by cutting the tip of the mouse's
tail, which no one in group 3 had ever done, so there were still feelings of fear or
nervousness during the process of taking blood samples. Another obstacle in group
3 was that the members did not know how to stop the flow of blood coming out of
the tip of the mouse's tail so that blood continued to come out of the tip of the
mouse's tail, but over time the blood stopped by itself. The final obstacle was that
after taking blood samples, the mice became a little more aggressive and agile. This
is thought to be caused by the fear that arises in mice during the process of taking
blood samples.
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VII. CONCLUSION

In this practicum it can be concluded that:

In this experiment, mice were given Glibenclamide, an antidiabetic drug,

after administration of dextrosal to raise blood sugar levels. The results showed a
reduction in blood sugar levels from 188 mg/dl to 89 mg/dl, indicating that
Glibenclamide had succeeded in reducing the blood sugar levels of mice. This
shows the effectiveness of the drug in controlling diabetes. In contrast, group 4 who
used Metformin as an antidiabetic drug saw an increase in blood sugar levels from
143 mg/dl to 165 mg/dl. This may be caused by errors in the experimental
procedures or inappropriate drug dosage. This shows the importance of proper
procedures and accurate dose determination in research.From the results of the two
groups, contradictory results were obtained, using glibenclamide reduced blood
glucose levels in mice, while metformin, on the contrary, increased blood glucose
levels. Frompractice,MetformindoesnotmatchtheliteraturebecauseMetformin is
supposed to be a good drug in lowering blood glucose levels. So from these results the
drugthatworksappropriatelyisglibenclamide.

VIII. BIBLIOGRAPHY

Soermaji, A. A. (2004). Rapid Determination of Blood SugarLevels in Mice: to be

Applied in Screening Antidiabetic Activity in Vivo. 115-116.

Wati, A. (2014). Comparison of the Hypoglycemic Effectiveness of the Patented

and Generic Metformin Drugs With the Logo Based on Allocation-Induced
Reduction in Blood Glucose Levels of Male Mice(Mus musculus) . AS-
SYIFAA, 91-97.

Widyastuti, S. (2022). Test the Antidiabetic Effectiveness of a Combination of

Senggani Leaf Extract and Glibenclamide in Reducing Blood Glucose
Levels in Mice. SCIENCE AND HEALTH JOURNAL, 262-267.
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IX. ATTACHMENT

Picture 1: Weighing of the Mice

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Picture 2: Oral Administration Dextrose

Picture 3: After Dextrose Administration

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Picture 4: Cut off Part of the Tail

Picture 5: Blood Sampling

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Picture 6: Result After Dextrose

Picture 7: Result After Glibenclamide

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Picture 8: Result Group 4 After Dextrose

Picture 9: Result Group 4 After Metformine

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X. QUESTION

1. What is meant by Diabetes Mellitus?

Diabetes mellitus is a syndrome characterized by high blood sugar (hyperglycemia)

due to impaired insulin production, secretion or insulin resistance. DM is very
important because of the complications it causes. Chronic complications of DM are
mainly based on vascular abnormalities, namely small blood vessels
(microangiopathy) and large blood vessels (macroangiopathy). Microangiopathy
manifests, especially in diabetic reitnopathy which can result in blindness, in the
kidneys diabetic nephropathy occurs which can ultimately result in kidney failure.
Macroangopathy can manifest in the lower limbs which can facilitate the
development of diabetic gangrene which may require amputation.

2. Name several types of Diabetes Mellitus and explain them!

Type 1 DM: DM caused by a complete absence of insulin production.

Type 2 DM: DM caused by insufficient and ineffective insulin action. Gestational

DM: DM that occurs during pregnancy

Other types of DM: Other types of DM caused by drug use, other diseases

3. How does Diabetes Mellitus occur?

The process of transporting glucose from the blood into the cells is carried out by
the hormone insulin produced by the pancreas. Simply put, insulin functions as a
regulator of blood sugar levels. People with diabetes cannot make insulin or cannot
respond to insulin well (insulin resistance). As a result, the transport of glucose into
cells becomes inadequate so that glucose accumulates in the blood and we can see
this through examination results of high blood glucose levels.

4. Name the drug classes for Diabetes Mellitus!

- Metformin
- Sulfonylurea
- Meglitinide
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- Thiazolidinediones
- DPP-4 inhibitors
- SGLT2 inhibitors
- GLP-1 receptor agonist (Incretin Mimetic)
- Alpha-Glucosidase Inhibitor
- Insulin