Antidiabetes effect of Noni Fruit (Morinda citrifolia L.)
on mice with Oral Glucose Tolerance Method and Streptozotocin Induction Method
1Faculty of Pharmacy,
Universitas Padjadjaran, Indonesia.
2Faculty of Pharmacy, Mandala Waluya, Health Science Academy,
Indonesia.
3Faculty of Pharmacy, Halu
Oleo University, Indonesia.
*Corresponding Author
E-mail: nikeherpianti.apt@gmail.com
Diabetes mellitus (DM) is a problem
that deserves attention because of its increasing prevalence every year. The prevalence of DM based on WHO in 2030 is
predicted to reach 366 patients. Epidemiologically, it is estimated that in 2030 the prevalence of DM in
Indonesia reaches 21.3 million people. The purpose of this study was to determine the effectiveness of glycosides
from noni fruit on reducing blood sugar levels in normal rats induced by diabetes by the method of glucose
tolerance and induction of streptozotocin (STZ). Antidiabetic effect testing was divided into 8 groups namely 4 groups
for oral glucose tolerance test in male mice (positive control group, negative
control group, glycoside control
group, and normal group), 4
other groups for testing
with STZ induction (positive control group,
positive control group, negative control, glycoside control group, and normal group). Oral glucose tolerance test
results on normal mice showed that glycosides from noni fruit (Morinda citrifolia L.) gave significantly different results
with negative controls at minute 30 is 0.036 (p <0.05) and were not significantly different from positive
controls (0.462) subsequently at 120 minutes the results showed that the glycoside group was significantly different
from the negative group (0.028) and not significantly different from the positive group (0.261). Tests with STZ
induction method showed that the decrease in the level of blood sugar induced
by the glycoside group was not significantly different (p> 0.05) with the positive group on day 1 (0.056), day 3 (0.168), and day 7(0.141)
so that it could be concluded that the glycosides from Noni fruit with a dose of
150mg/kg body weight provides antidiabetic activity.
KEYWORDS: Glycosides, Morinda citrifolia, Diabetes
mellitus, Oral glucose
tolerance, Streptozotocin (STZ).
The threat of Diabetes Mellitus (DM)
in the midst of people's lives is a very frightening specter because almost every 10 seconds in the world
people die due to complications from
the disease. DM is a chronic disease that occurs when the body cannot produce enough insulin
or when the body does not use insulin effectively which results in a metabolic
disease characterized by chronic hyperglycemia and abnormalities in carbohydrate, protein,
and fat metabolism1.
One
of the goals of therapy
for patients with DM is controlling blood sugar levels by administering oral hypoglycemic drugs or exogenous
insulin. Pharmacological
therapy of DM is generally given in the long
term so that this can increase the risk of side effects from medications such as hypoglycemia, liver and kidney damage, psychonia, and lactic acidosis.
This patient safety factor is a consideration for finding alternatives in dealing with DM from
natural ingredients. The use of
medicinal plants and phytochemicals to treat
DM is to look for safer alternatives as medicines, which temporarily reduce blood glucose,
prevent heart disease and high blood pressure, and also improve
antioxidant systems, insulin action
and secretion2
The prospect of using natural
products to treat DM has not been
much studied. One of the medicinal plants used
to reduce blood glucose levels is noni (M. citrifolia
L). Noni (M. citrifolia L.) contains
polysaccharides, glycosides, flavonoids, iridoids, carotinoids, and anthraquinones3. Flavonoids
that function as antioxidants are able to withstand
the rate of absorption of blood glucose from the digestive tract to
blood vessels so that they are able to
withstand the rate of increase in blood glucose
levels4. Flavonoids are divided into 5 sub-classes including flavons,
flavonols, flavonons, isoflavons, and anthocyanins5.
The sub-class of flavonols is subdivided, one of which consists of quercetin compounds.
Several studies have focused on
quercetin to be developed as an antidiabetic drug to
prevent and manage DM.
Previous studies have reported
quercetin mechanisms in DM cases, such as decreased lipid peroxidation, increased antioxidant enzymes (SOD, GPX,
and CAT), reduced glucose absorption
in the intestine by inhibiting GLUT26,7 .
Research conducted by Kwon et al, (2007) explained
that the transport of fructose and glucose by
GLUT2 is strongly inhibited by quercetin.
Separation of glycoside compounds
was carried out by the process
of extraction of the sympathetic by maceration method to obtain maserate. Maserate is obtained thickened using a rotary
evaporator, then the thick extract is
added with ethyl acetate to separate the glycosides
from the Morinda
citrifolia L.
extract with the partitioning
process, the glycosides from the Noni fruit
will be at the bottom and the top is the ethyl extract layer. acetate. The separated glycosides are then thickened.
The experimental animals used in
this study were Wistar strain male
mice aged 8 weeks and weighed between 20- 30g
and placed in separate cages according to the test group. The animals were adapted
in the experimental cage one week before being treated. The
state of the cage is maintained at
a temperature of 28-32°C and a dark- light
cycle of 12 hours each. Experimental animals were fed standard diet pellets and ad libitum
drinking water8,
10. The
total volume of administration is 1
ml which is the volume that can be given based on the normal volume of the stomach of mice8.
In this test the test animals were
divided into 8 groups, 4 groups for the oral glucose tolerance
test and 4 groups
for the test in STZ induced mice. Each group consists of 3 mice.
Test animals are divided into 4 groups, consisting of
: Group I :Negative Control (Na. CMC 1%)
Group II :Positive Control (Glibenclamide 5mg/kg BW)
Group III :Extract Control (Noni glycosides 150mg/ kgBW)
Group IV :Normal control
(sufficient food and drink)
Test animals are divided into 4 groups,
consisting of :
Group 1
:Negative Control (Na. CMC 1%)
Group 2 :Positive Control (Exogenous Insulin)
Group 3 :Extract Control (Noni glycosides 150mg/ kgBW)
Group 4 :Normal control (sufficient food and drink)
On the first day before treatment
all treatment groups were fasted for
20 hours. After 20 hours blood sugar levels were measured from each group. Then each treatment group was induced
by glucose orally,
then blood glucose levels were
measured again in the 30th minute.
Furthermore, they are given treatment according to their respective groups. Blood sugar levels were measured
at 0, 30, 60, 90, 120, 150, 180 minutes.
On
the first day before treatment
all rats were fasted for
20
hours, then their fasting blood sugar levels were examined. Induction of diabetes in
experimental animals is done by giving
STZ (45mg/kg BW) intraperitoneally. Blood
sugar levels of mice were checked again on the
second day, 24 hours after STZ injection. Mice are declared diabetic if they have glucose
levels of 200-260 mg/dL, but this did
not happen in this study which can be
caused by several factors including lowering fasting blood sugar levels
of experimental animals
or the level of stress
experienced by experimental animals1,8. After that, the test animals are given treatment according to their respective groups.
The
results of the study are
expressed as mean±SEM. The significance of the data was analyzed by One-way Analysis of Variance (ANOVA) (SPSS 16.0
program) with post hoc LSD's test. Data is considered
significant if the p value
is ≤ 0.05.
Fig 1: Glucose-Induced Blood Sugar
Level Diagram of Mice
|
Ket. |
: KGDP |
: Blood glucose level during fasting (mg / dL) |
|
|
KGDG |
: Blood glucose Level After induced Glucose (mg / dL) |
|
|
0 |
: Blood glucose Level after Treatment at 0 minute (mg / dL) |
|
|
30 |
: Blood glucose Level after Treatment at 30 minute (mg / dL) |
|
|
60 |
: Blood glucose Level after Treatment at 60 minute (mg / dL) |
|
|
90 |
: Blood glucose Level after Treatment at 90 minute (mg / dL) |
|
|
120 |
: Blood glucose Level after Treatment at 120 minute (mg / dL) |
|
|
150 |
: Blood glucose Level after Treatment at 150 minute (mg / dL) |
|
|
180 |
: Blood glucose Level after Treatment at 180 minute (mg / dL) |
Table 1. Glucose-Induced Blood
Sugar Level Diagram
of Mice
|
Observational result |
Treatments |
IK 95% |
|
|
Mean diffrence |
P |
||
|
KGDSIG |
Negative vs Normal |
72.000* |
0.000 |
|
Negative vs Glikoside |
1.333 |
0.893 |
|
|
Negative vs Positive |
14.000 |
0.184 |
|
|
Positive vs Glikoside |
-12.667 |
0.225 |
|
|
Minute 0 |
Negative vs Normal |
77.667* |
0.000 |
|
Negative vs Glikoside |
8.667 |
0.380 |
|
|
Negative vs Positive |
15.667 |
0.131 |
|
|
Positive vs Glikoside |
-7.000 |
0.474 |
|
|
Minute 30 |
Negative vs Normal |
137.000* |
0.011 |
|
Negative vs Glikoside |
105.000* |
0.036 |
|
|
Negative vs Positive |
137.333* |
0.011 |
|
|
Positive vs Glikoside |
-32.333 |
0.462 |
|
|
Minute 60 |
Negative vs Normal |
86.333* |
0.011 |
|
Negative vs Glikoside |
60.333* |
0.049 |
|
|
Negative vs Positive |
134.000* |
0.001 |
|
|
Positive vs Glikoside |
-73.667* |
0.022 |
|
|
Minute 90 |
Negative vs Normal |
79.000* |
0.008 |
|
Negative vs Glikoside |
63.333* |
0.023 |
|
|
Negative vs Positive |
88.333* |
0.005 |
|
|
Positive vs Glikoside |
-79.000* |
0.008 |
|
|
Minute 120 |
Negative vs Normal |
64.667* |
0.030 |
|
Negative vs Glikoside |
65.667* |
0.028 |
|
|
Negative vs Positive |
95.333* |
0.005 |
|
|
Positive vs Glikoside |
-29.667 |
0.261 |
|
|
Minute 150 |
Negative vs Normal |
77.667* |
0.002 |
|
Negative vs Glikoside |
81.000* |
0.001 |
|
|
Negative vs Positive |
86.000* |
0.001 |
|
|
Positive vs Glikoside |
-5.000 |
0.770 |
|
|
Minute 180 |
Negative vs Normal |
129.667* |
0.000 |
|
Negative vs Glikoside |
125.000* |
0.000 |
|
|
Negative vs Positive |
139.000* |
0.000 |
|
|
Positive vs Glikoside |
-14.000 |
0.475 |
|
Fig 2: STZ Induced Blood Sugar Level
Diagram of Mice
|
Ket. |
: KGDP |
: Blood glucose level during fasting (mg/dL) |
|
KGDSISTZ |
: Blood glucose Level 24 hours after STZ Induction (mg/dL) |
|
|
Day 1 |
: Blood glucose Levels H + 1 Treatment (mg/dL) |
|
|
Day 3 |
: Blood glucose Levels H + 3 Treatment (mg/dL) |
|
|
Day 7 |
: Blood glucose Levels H + 7 Treatment (mg/dL) |
Table 2. STZ Induced
Blood Sugar Level Diagram of Mice
|
Observational result |
Treatments |
IK 95% |
|
|
Mean diffrence |
P |
||
|
KGDSISTZ |
Positive vs Negative |
-28.000 |
0.222 |
|
Positive vs Normal |
279.333* |
0.000 |
|
|
Positive vs Glikoside |
-2.000 |
0.927 |
|
|
Negative vs Glikoside |
26.000 |
0.254 |
|
|
Day 1 |
Positive vs Negative |
-206.667* |
0.000 |
|
Positive vs Normal |
96.000* |
0.002 |
|
|
Positive vs Glikoside |
-17.333 |
0.424 |
|
|
Negative vs Glikoside |
-113.333* |
0.001 |
|
|
Day 3 |
Positive vs Negative |
-191.333* |
.000 |
|
Positive vs Normal |
69.333* |
.000 |
|
|
Positive vs Glikoside |
-4.000 |
.610 |
|
|
Negative vs Glikoside |
187.333* |
.000 |
|
|
Day 7 |
Positive vs Negative |
-405.667* |
.000 |
|
Positive vs Normal |
-47.000 |
.054 |
|
|
Positive vs Glikoside |
-32.667 |
.155 |
|
|
Negative vs Glikoside |
373.000* |
.000 |
|
Normality test carried out on each
treatment group after giving glucose and after treatment minutes 0, 30, 60, 90, 120, 150, and 180 using the
Shapiro-Wilk test, showed that at all times the treatment
data was normally
distributed. The analysis
shows that there is no real difference. The result seen in figure 1.
Statistical results on the oral glucose
induction showed that the blood sugar levels induced by the glycoside
group to the negative and positive groups were not significantly different (p> 0.05) with significant
values of 0.036 and 0.462, respectively, this proved that all induced
experimental animals had hyperglycemia, differing the case with the normal group without glucose
induction showed significant results
in the negative group, positive group,
and glycoside group. This is also in line with the statistical results of the induction of blood sugar levels in the STZ
group. The result can be seen in figure
2.
This research aims to look at
antidiabetic activity in mice test animals
with 2 methods namely to prove that glycoside compounds
from noni fruit can act as antidiabetic in diabtes
model animals and normal test animals.
From the results it can be concluded that the
glycoside compound testing
using the oral glucose tolerance test method provides an optimal
antidiabetic effect by proving that
the measurement of blood glucose levels decreased
blood sugar levels by 20.03% and positive control by 49.62%. This is also
supported by statistical data based
on the post hoc test ie at
the 30th minute, the decrease in blood sugar levels of the glycoside group (0.716) was not
significantly different (p> 0.05) with the positive group (0.490) while the negative
control group showed a significant result of 0.032.
This
research above, in line with some previous studies where it is said that the compounds contained in noni fruit have antidiabetic properties. The compounds
contained in noni are ursolic acid (triterpenoids); caprylic
acid and hexanoic
acid (fatty acid); niacin; asperulosidic acid (iridoids); routine
and quercetin (flavonoids); 2,6-di-O- (β-D12 glucopyranosyl 1-O- octanoil-β-D glucopyranose (fatty acid esters);
damnacanthal
(antrakinone); americanin A (lignans), xeronin
(alkaloids) and scopoletin (coumarin) (fatty acid esters);
damnacanthal
(antrakinone); americanin A (lignans); xeronins (alkaloids) and scopoletin
(coumarin) (fatty acids)9,10,11.
In the STZ induction testing group, normality testing
using the Shapiro-Wilk test showed normal
distributed data (p> 0,05).
Tests on the STZ induction group performed
showed a significant difference in the H + 3 treatment of the negative
control group to the normal and glycoside
treatment groups. From these data it is known that the glycoside compound provides antidiabetic
effect because the results
are not significantly different from the positive group ie exogenous insulin. This is also in line with the results of blood sugar level
measurements on day 7. Antidiabetic
activity is also indicated by a percent decrease in the control
group. positive 63.48% and glycoside
compound group by 62.36% while for the negative group there was no decrease
in blood sugar
levels from induced
blood sugar levels.
The results of this study indicate
that glycosides which are the largest constituent of the components of the metabolite compounds in noni fruit provide
effectiveness for healthy
animals and diabetes.
Based on the results of the study
after statistical analysis and
discussion, it can be concluded that:
1. Glycosides at a
dose of 150mg/kg BW from noni extract (Morinda citrifolia L.) showed antidiabetic activity in test animals with oral glucose tolerance
test methods and were not significantly different
from positive control activities.
2.
Glycosides
at a dose of 150mg/kg BW from noni extract (Morinda citrifolia L.) showed antidiabetic activity in test animals using STZ induction method and not significantly different from
positive control activities.
The
authors wish to express gratitude
to STIKES Mandala
Waluya for support
to implementing this study
There are no conflicts of interest.
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Received on 07.07.2020
Modified on 14.11.202
Accepted on 17.01.2021
© RJPT All right reserve
Research J. Pharm. and Tech. 2021; 14(10):5067-5071.
DOI: 10.52711/0974-360X.2021.00883