1. EFFECT OF Syzygium polyanthum ETHYL ACETATE FRACTION AGAINST LANGERHANS ISLETS
HISTOPATHOLOGICAL CHANGES IN RATS WITH ALLOXAN-INDUCED
DIABETES MELLITUS
By:
Nabilla Asmara Dewi Edyta
061611133032
Supervisor Co-Supervsor
(Dr. Eka Pramyrtha Hestianah., drh., M.Kes.) (Prof. Dr. Dewa Ketut Meles, drh., MS.)
Head Examiner Secretary Examiner Member
(Suryo Kuncorojakti, drh., M.Vet., Ph.D.) (Dr. Nusdianto Triakoso, MP., drh.) (Dr. Rochmah Kurnijasanti, M.Si., drh.)
2. Background
Diabetes mellitus (DM) belongs to group of metabolic
diseases in which the main symptom associated with this disease is
the high blood sugar levels for a long period of time (Mehmet et
all., 2017).
The increase of ROS generation causes oxidative stress,
including several cellular changes. Acute or chronic high glucose
in diabetes increases ROS production and activates apoptosis in
the β-cells (Volpe, 2018).
The use of OHAs is expensive along with the side effects.
Apart from being used as spices, S. polyanthum can also be used as
medicine. Bay leaves contain essentials oil (consisting of citrol,
triterpenoid, saponins, flavonoids and tannins). Bay leaves also
contain vitamin A, vitamin C, vitamin E, vitamin B6, vitamin B12,
thiamin, riboflavin, niacin, and folate (David and Branen, 1993).
3. Problem Formulation
Does the administration of bay leaves (Syzygium
polyanthum) fraction affect the number and the structure
of β-cells of Langerhans islets in rats with alloxan-
induced diabetes mellitus?
4. Research Purpose
Knowing the effects of bay leaves (Syzygium
polyanthum) fraction that were administered orally to
white rats (Rattus norvegicus) as antidiabetic treatment
and to know the histopathological changes of the
Langerhans islets in rats with alloxan-induced diabetes
mellitus.
5. Benefits of Research
• This study is aimed to prove bay leaves (S. polyanthum) towards blood sugar levels
and Langerhans islets of hyperglycemic rats (Rattus norvegicus). The results of this
study are expected to provide additional information about the potential of bay leaves
fraction (Syzygium polyanthum) as an alternative treatment for diabetes mellitus, in
increasing insulin level by repairing the α-cells and β-cells of Langerhans islets
1st Theoretical Benefits
• The results of this study are expected to increase the economic value of bay leaves
for use as an alternative antidiabetic drug.
2nd Practical Benefits
6. Theoretical Basis
This study used alloxan as diabetogenic agent. Alloxan
inhibits many cellular function at higher concentrations
such as the ability to oxidize many thiol groups namely
hexokinase, phosphofructokinase, calmodulin-dependent
protein kinase, acotinase and other protein. Biological
effect of alloxan is pancreatic β-cells toxicity and
diabetogenicity that may be attributed to alloxan-induced
redox cycling and ROS.
7. Hypothesis
The administration of bay leaves (Syzygium polyanthum)
fraction affect the number and the structure of β-cells of
Langerhans islets in rats with alloxan-induced diabetes
mellitus.
8. Research Method
Independent variable in this research are bay leaves (Syzygium polyanthum) fraction
concentrations.
Independent Variable
Dependent variable in this research are the histological features of Langerhans islets.
Dependent Variable
The control variable of this research is the experimental animals, sex, age, weight,
maintenance, treatment, and the cages are in the same condition.
Controlled Variable
9. Location and Date of Research
• This research was carried out in Laboratory of
Pharmacology
• This research was conducted around May 2023
10. Research Sample
The number of rats was determined from the number of
replications, which for experimental research with a
complete randomized, randomized group or factorial
design. This study requires 6 treatment groups and the
number of replications of 4.
Criteria:
• The rats age around 2-3 months old
• Weigh around 180-200 grams
• Never been used as animal laboratory before
• Males rats
• In healthy condition
11. Material and Tools
Research Materials:
• Bay leaves (Syzygium
polyanhtum)
• Methanol
• Etyl acetate
• NH4OH
• CMC Na 0,5%
• Alloxan
• Metformin
• Aquades
• NaCl
Research Tools:
• Cages
• Masker
• Gloves
• Vacuum pump
• Evaporator
• Buchner funnel
• Erlenmeyer glass
• Separatory funnel
• Glucomter
• Minor surgical set
• Microscope
• Oral gavage
• Syringe 3CC
12. Flow Chart
K-
CMC Na
0,5% +
Bay leaves
fraction
500
mg/kgBW
CMC Na
0,5% +
Bay leaves
fraction
125
mg/kgBW
CMC Na
0,5% +
Bay leaves
fraction
250
mg/kgBW
CMC Na
0,5%
CMC Na
0,5%
On day 9th, induct alloxan using 170
mg/kgBW interperiotally
Rats euthanized, pancreas collection and fixation
using Buffer Phosphate Formalin 10%
24 rats adapted for 7 days
K+ T2
T1
T0 T3
On day 8th, rats were fasted and blood sugar
levels were measured before given alloxan
Normal blood sugar
level 50-135 mg/dL
Group tratment
CMC Na
0,5% +
metform
in 45
mg/kgB
W
Blood sugar levels measurement on day 14th post therapy
Histological slides preparation with Mallory Azan
staining
Histopathological changes observation
Data analysis
14. Research Result
The average number of pancreatic β-cells
Treatment Mean ± SD
C+ 32.27b ± 8.48
C- 29.74b ± 7.67
T0 9.31a ± 2.60
T1 18.11ab ± 9.76
T2 31.83b ± 10.48
T3 27.39b ± 13.97
15. Research Result
The scoring results of damage on Langerhans islets
Treatment Mean ± SD
C- 0.33b ± 0.5
C+ 1.00b ± 0.0
T0 3.75a ± 0.5
T1 3.5ab ± 0.57
T2 1.00b ± 0.00
T3 1.5b ± 0.57
16. Conclussion
• Bay leaves fraction at doses 125, 250, and 500 mg/kgBW are
effective to repair the damage of Langerhans islets. Bay leaves
fraction increased the number of pancreatic β-cell after the
alloxan induction.
• Bay leaves fraction repaired the structure of Langerhans islets
after the alloxan induction.
17. Suggestion
• Further research on the results of the isolation of antioxidant
compounds of bay leaves fraction bioactivity as an antioxidant
should be tested.
• The isolation treatment of other compounds from other active
fractions needs to be studied more carry on. This was done to
look for potential compounds and can be used as a drug with in
vivo clinical trials for know the value of toxicity and has a good
level of safety.
