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Eduvest � Journal of
Universal Studies Volume 4
Number 11, November, 2024 p- ISSN
2775-3735- e-ISSN
2775-3727 |
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THE EFFECT COMBINING SNAKEHEAD
FISH EXTRACT, MENIRAN, AND TEMULAWAK ON GLYCEMIC STATUS AND PANCREATIC
HISTOPATHOLOGY IN HIGH-FAT DIET DIABETIC RATS |
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Wa Amalia Zahiah1, OK Yulizal2,
Erwin Sopacua3 1,2,3Faculty of Medicine, Universitas Prima Indonesia, Indonesia Email: [email protected]1, [email protected]2, [email protected]3 |
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ABSTRACT |
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Diabetes mellitus is a metabolic
disorder characterized by hyperglycemia, frequently associated with obesity,
dyslipidemia, hypertension, and cardiovascular complications, particularly in
type 2 diabetes mellitus (DM). In Indonesia, traditional medicine is often
favored for its safety and cost-effectiveness compared to synthetic drugs.
This study involved 25 male Wistar rats, which were divided into five groups:
a control group, an alloxan and high-fat diet group, a pioglitazone group, a
plant extract group, and a combination group. Obesity was induced over 27
days through a high-fat diet, followed by the administration of alloxan to
elevate blood glucose levels. Glucose measurements were taken at specified
intervals, with HbA1c assessed on day 26, and pancreatic histopathology was
analyzed post-study. The Kruskal-Wallis test revealed significant differences
in weight gain (p = 0.016) and blood glucose levels (p = 0.003) among the
groups. Although no significant difference was observed in blood glucose
reduction (p = 0.05), the combination group exhibited the most substantial
decrease. One-way ANOVA results demonstrated that the combination of the
extract and pioglitazone significantly reduced HbA1c levels (p < 0.001),
with the second group showing the highest levels and the fourth group
displaying the most pronounced reduction. Histopathological analysis
indicated damage to the islets of Langerhans in both the pioglitazone and
extract groups. The combination of snakehead fish extract, meniran,
temulawak, and pioglitazone effectively lowers blood glucose and HbA1c
levels; however, it does not appear to ameliorate islet damage, likely due to
oxidative stress resulting from the treatments. |
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KEYWORDS |
Diabetes
mellitus , Snakehead Fish Extract, Meniran, Temulawak, Glycemic Status,
Pancreatic Histopathology. |
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This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International |
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The pancreas is an accessory organ in the
digestive system that is located in the Retroperitoneal and is flattened like a
sponge, measuring about 12 to 15 cm long and 2.5 cm thick. The pancreas comprises
three parts: a head surrounded by a duodenum, a central or torso, and a blunt
and pointed tail on the left. The pancreas acts as exocrine and endocrine
glands. The islands of the pancreatic Langerhans tend to be more abundant in
the tail of the pancreas, while the head of the pancreas is dominated by
exocrine tissue. About 99% of the pancreatic structure is exocrine tissue that
secretes 1,200 to 1,500 mL/day of pancreatic fluid. In the endocrine part, the
island of Langerhans pancreas produces insulin and glucagon (Saladin et al., 2010).
Diabetes mellitus (DM) is a group of common
metabolic disorders in which hyperglycemia is the main feature. The variation
of the type of diabetes mellitus is influenced by the complex interaction
between genetic and environmental factors. Diabetes mellitus is a condition in
which there is damage to the cells of the pancreatic Langerhans (in the form of
necrosis and vacuolisation), thus indicating damage or breakdown in the beta (Jameson, 2010) cells of the Langerhans.
In this state, lymphocytes can penetrate into the pancreatic Langerhans, thus
indicating the presence of an autoimmune process against the Langerhans beta (β) cells. Based on the causes, factors that trigger hyperglycemia include
decreased insulin secretion, inefficient use of glucose, and increased glucose
production (Jameson, 2010) (Margaretha, 2016).
Diabetes mellitus is closely related to
obesity, specifically type 2, and is a risk factor for dyslipidemia,
hypertension, as well as heart and vascular disease, which later becomes the
main complication and leading cause of death in individuals with type 2
diabetes mellitus. In addition, a lifestyle that lacks physical activity,
leading to overweight and obesity, is one of the factors that can be changed in
the occurrence of diabetes mellitus. Other risk factors such as an unbalanced
diet, a history of impaired glucose tolerance, or disturbances in fasting blood
glucose levels (Ardiani et al., 2021) (N. N. Sari, 2018).
In type 1 diabetes mellitus, pancreas's the
beta (β) ceeas are damaged by autoimmune processes, resulting in the absence of
insulin production. This condition leads to fasting hyperglycemia, in which the
liver produces glucose without proper regulation. Although glucose from food
circulates in the blood and causes postprandial hyperglycemia, the liver
is unable to store that glucose. When blood glucose levels are very high, the
kidneys are unable to reabsorb the glucose that has been filtered, causing
glucose to appear in the urine, known as diabetes. The osmotic diuresis process
is the process of excretion of excess glucose along with excess electrolytes,
which results in increased frequency of urination (polyuria) and excessive
thirst (polydipsia) (Lestari & Zulkarnain, 2021).
In type 2 diabetes mellitus, insulin
resistance in muscle and liver cells and pancreatic beta cell dysfunction are
the central damaging pathophysiology of type 2 diabetes mellitus. Recent
research suggests that pancreatic beta cell failure occurs earlier and is more
severe than previously thought. Other organs that participate in type 2
diabetes mellitus are adipose tissue (experiencing increased lipolysis),
gastrointestinal (experiencing incretin deficiency), alpha cells of the
pancreas (experiencing hyperglycagonia), kidneys (experiencing increased
glucose absorption), and brain (experiencing insulin resistance) thus causing
impaired glucose tolerance. Currently, three new pathogenesis pathways have
been identified in the concept of "ominous octet," which plays
a role in the occurrence of hyperglycemia in type 2 diabetes mellitus (Indonesia, 2021).
Patients with diabetes mellitus need a
variety of treatments to reduce the risk of micro and macrovascular
complications. Drugs from the thiazolidindione group, such as
pioglitazone, can be used as one of the options in the treatment of diabetes
mellitus. This drug works by increasing sensitivity to insulin so that it is
able to overcome insulin resistance problems and its complications without
causing hypoglycemia. However, the use of this group of drugs can cause side
effects such as edema, weight gain, heart failure, and risk of fractures. (Lebovitz, 2019) (Malihah & Emelia, 2022)
In Indonesia, many people prefer the use of
traditional medicine as a method of prevention and treatment of various
diseases, including diabetes mellitus (DM), rather than using synthetic drugs.
The reason is that traditional medicines are claimed to have fewer side
effects, are affordable, and are easy to get. Traditional medicine can be in
the form of ingredients or mixtures of plants, animals, minerals and galenic
preparations that have been used for generations to overcome various diseases,
including diabetes mellitus (Anam et al., n.d.).
One of the natural ingredients that can be
used for the treatment of diabetes is a combination of snakehead fish (Channa
striata), meniran Phyllanthus niruri L.), and temulawak (Curcuma
xanthorrhiza), which are available in finished capsule preparations. Channa
striata extract is known as a source of animal protein that is rich in
essential nutrients, which plays an important role in increasing the body's
stamina after childbirth, surgery, and the recovery process from certain
diseases, and has anti-inflammatory, antioxidant and protective abilities
against peptic ulcers (Yulizal et al., 2020) .α −glucosidase. This enzyme is responsible for converting
carbohydrates into glucose. Thus making it effective in helping to control
glucose levels in the blood (Soniya & Fauziah, 2020).
Temulawak (Curcuma xanthorrhiza) is a
plant native to Indonesia that has the potential to be an antidiabetic.
Secondary metabolites contained in curcuma are believed to reduce blood glucose
levels. In addition,
Green Meniran (Phyllanthus niruri L.)
is a plant that is known to have effects that can increase the body's
immune system or immunostimulant. Its main contents include flavonoids,
phyllants, hypophylantines, resin, and tannins. These ingredients are
believed to be beneficial as diuretics, antioxidants, anti-inflammatory,
antidiabetic, and antipyretic, and can increase appetite. In medical
activities, this plant is used to treat various conditions, such as kidney
stones, dyspepsia, hepatotoxicity, and has the potential as an
antihyperglycemic agent. However, studies on the antihyperglycemic and
antioxidant potential of meniran (H. Sari et al., 2019). (Phyllanthus niruri
L.) is still fairly rare (Kumar et al., 2019).
Based on the description above, extracts from
snakehead fish (Channa striata), meniran (Phyllanthus niruri L.),
and temulawak (Curcuma xanthorrhiza) show potential as
antihyperglycemic agents in the treatment of diabetes mellitus. Based on these
findings, researchers became interested in further studying the effects of
animal and plant extracts formulated in combination capsule preparations as an
alternative treatment for diabetes mellitus.
RESEARCH METHOD
This
study is an experimental research with a randomized post-test-only control
group design. The object of the study used was male rats of the Wistar strain
(Rattus norvegicus) induced by aloxan to assess the glycemic status and
histopathological picture of the pancreas. This research has received an
ethical permit with the number 046/KEPK/UNPRI/III/2024.
The
research data source consisted of 30 male rats of the Wistar strain aged 6-8
weeks with a body weight of 150-250 grams, obtained from the Ellio Medan
Laboratory. The study population was male rats of the Wistar strain, while
samples were taken based on strict inclusion and exclusion criteria. The
inclusion criteria included healthy mice, had no anatomical abnormalities, and
had not been used in previous studies. Exclusion criteria included mice that
died during the study or were not actively moving. The sample size was
calculated using Federer's formula, resulting in the need for a minimum of 5
rats per treatment group, which was then rounded to 6 rats per group to anticipate
the death of rats.
Research
techniques and tools include providing a high-fat diet to induce diabetes and using
capsules with a combination of snakehead fish extract, temulawak, and meniran,
as well as pioglitazone as a positive control. Blood glucose levels were
measured using a glucometer, while HbA1c levels were measured using an
i-CHROMA� device. Pancreatic histopathology preparations were made using the
paraffin technique, stained with Hematoxylin and Eosin (HE), and examined using
the Olympus Cx-21 microscope.
Data
analysis was carried out using the SPSS version 29 program. The data obtained
were tested for normality using the Shapiro-Wilk test. For the analysis of the
effect of the combination of extracts on the glycemic status of mice, a variant
analysis (ANOVA) was used if the data were normally distributed (P > 0.05).
In contrast, the Kruskal-Wallis H test was applied to the normally
undistributed data (P < 0.05), in order to determine the mean difference in
results between each treatment group.
RESULT AND DISCUSSION
Effect of High Fat Diet in Rats
The
study showed that mice in the group before and after the treatment experienced
weight gain as a result of feeding a high-fat diet. Each group experienced an
average weight gain of 10 grams. The following table shows the complete data on
the weight changes:
Table
1. Shapiro-Wilk Test of Rat Body Weight
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Treatment Groups |
Mean � SD |
P value |
Distribution Data |
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Initial Weight Loss |
K-1 |
151.20 � 1.30 |
0,421 |
Usual |
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K-2 |
151.80 � 1.92 |
0,223 |
Usual |
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K-3 |
152.00 � 0.70 |
0,325 |
Usual |
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K-4 |
150.40 � 0.54 |
0,314 |
Usual |
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K-5 |
151.80 � 1.23 |
0,006 |
Abnormal |
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Weight Loss After Treatment |
K-1 |
153.80 � 2.07 |
0,656 |
Usual |
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K-2 |
164.80 � 0.83 |
0,314 |
Usual |
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K-3 |
162.60 � 5.77 |
0,866 |
Usual |
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K-4 |
164.60 � 4.50 |
0,957 |
Usual |
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|
K-5 |
164.40 � 1.67 |
0,314 |
Usual |
The results of the
Shapiro-Wilk test, based on the data above, show that certain groups have a p
< value of 0.05 while other groups have a p > value of 0.05. This
indicates an abnormal distribution of weight data. Therefore, the analysis
continued by applying the Kruskall-Wallis H non-parametric test to assess the
significant weight difference between body weight before and after the high-fat
diet treatment.
Table
2. Kruskal-Wallis Test of Rat
Body Weight
|
Parameters |
Group Treatment |
Weight (Gram) |
P value |
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Median |
Min |
Max |
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Initial Weight Loss |
K-1 |
150 |
150 |
150 |
0,128 |
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K-2 |
152,5 |
152 |
153 |
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K-3 |
152 |
151 |
153 |
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K-4 |
151 |
150 |
152 |
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K-5 |
150 |
150 |
151 |
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Weight Loss After Treatment |
K-1 |
153 |
150 |
157 |
0,016 |
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K-2 |
165 |
164 |
166 |
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K-3 |
162 |
155 |
171 |
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K-4 |
165 |
158 |
170 |
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K-5 |
164 |
162 |
166 |
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The
results of the Kruskal-Wallis test showed that the value on body weight after
treatment showed a p-value of 0.016 (< 0.05), which indicates a significant
influence of high-fat diet on the difference in body weight of male rats of the
Wistar strain (Rattus norvegicus). The weight of the rats in this study ranged
from 150 grams to 171 grams.
Effect of Aloxan Induction on Increase in KGD and Extract Administration on Decrease in KGD in Rats
The
results showed that blood glucose levels were in the normal range before naloxone
induction, after induction, and after administration. Each group showed
hyperglycemia conditions with glucose levels of more than 300 mg/dL and a
decrease in blood glucose levels after 14 days of administration of the extract
can be seen in the table below:
Table 3. Shapiro Wilk Test Results
for Each Treatment Group
|
Treatment Groups |
Mean � SD |
Shapiro Wilk (Sig.) Normality Test |
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Before |
After |
D+4 |
D+14 |
Before |
After |
D+4 |
D+14 |
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K-1 |
83 � 3.39 |
130.4 � 2.97 |
124.2 � 6.30 |
117.2 � 3.35 |
0.564 |
0.777 |
0.414 |
0.616 |
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K-2 |
80.8 � 5.63 |
469.2 � 90.6 |
466.6 � 132.12 |
399.4 � 82.18 |
0.272 |
0.169 |
0.129 |
0.341 |
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K-3 |
88.6 � 5.08 |
478.6 � 121.08 |
396.8 � 132.04 |
317.8 � 82.17 |
0.965 |
0.215 |
0.515 |
0.642 |
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K-4 |
89.6 � 8.08 |
402.0 � 128.18 |
374.4 � 85.05 |
212.8 � 143.73 |
0.548 |
0.914 |
0.957 |
0.018 |
|
K-5 |
92.0 � 5.83 |
428.8 � 69.08 |
322.4 � 113.4 |
107.2 � 11.12 |
0.351 |
0.669 |
0.674 |
0.002 |
The
above data showed that there was an increase in blood glucose levels in rats
after induction with aloxan 100 mg/kgBB, with an increase in blood glucose
levels of around 300 mg/dL in the treatment group that received aloxane. On the
14th day, treatment with a combination of snakehead fish extract (Channa striata), meniran (Phyllanthus niruri L.),
curcuma (Curcuma xanthorrhiza), and
pioglitazone lowered blood glucose levels in male rats of the Wistar strain (Rattus norvegicus) to below 100 mg/dL, as shown in the
data above. The analysis of glucose level reduction in five treatment groups
was carried out using� the Shapiro-Wilk test, with most of the
results of the p > value of 0.05, indicating normal distributed data, while
K-5 (Aloxan + high-fat diet +
pioglitazone on day 14) p < 0.05 showed abnormal data distribution.
Therefore,� the Kruskall-Wallis� test was
conducted to test for significant differences between the groups.
Table 4. Kruskal-Wallis Test After Administration
of H+14 Extract
|
Treatment |
Test Statistic (D+14) |
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Kruskall Wallis H |
Df |
Asym. Sig |
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K-1 |
16,315 |
4 |
0,003 |
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K-2 |
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K-3 |
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K-4 |
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K-5 |
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Based on the Kruskal-Wallis test, a p-value of
0.003 (< 0.05) was obtained; this value showed that there was a significant
difference in the average blood glucose level in male rats of the Wistar strain
(Rattus norvegicus) on D+14. To further examine the differences between
the treatment groups, a Mann Whitney post hoc test was performed, as seen in
the following graph:
a bc c d e
Figure 1. Graph of H+14 Glucose Level
Decrease
The
same letter notation showed that there was no difference in the reduction of
glucose levels between treatments, based on Mann Whitney's post-hoc test �at the singnifiability level of α = 0.05.
The K-5 treatment (aloxan + high fat diet + pioglitazone + extract)
showed the highest average decrease in glucose levels compared to other
treatments. This shows that the K-5 treatment is the most effective in lowering
blood glucose levels in the Wistar strain male rats (Rattus norvegicus). Thus,
it can be concluded that the combination of snakehead fish extract, meniran,
temulawak, and pioglitazone significantly lowers blood glucose levels.
Effectiveness of Snakehead Fish Extracts, Meniran, Temulawak, and Pioglitazone on HbA1c Levels in Wistar Rats
HbA1c measurements were performed to assess
glycemic control in people with diabetes mellitus. HbA1c was measured after the
mice underwent acclimatization, aloxane induction, and treatment with snakehead
fish extracts, meniran, temulawak, and pioglitazone for 26 days. The
examination of HbA1c levels in rat serum obtained the results in the following
table:
Table 5. HbA1c Levels in Mouse Serum
|
Group |
Result |
Units |
|
K1
(Normal) |
3.7 |
% |
|
K1
(Normal) |
3.6 |
% |
|
K1
(Normal) |
4.0 |
% |
|
K1
(Normal) |
3.5 |
% |
|
K1 (Normal) |
3.8 |
% |
|
K2
(Aloxan + CMC + High Fat Diet) |
9.3 |
% |
|
K2
(Aloxan + CMC + High Fat Diet) |
8.8 |
% |
|
K2
(Aloxan + CMC + High Fat Diet) |
8.9 |
% |
|
K2
(Aloxan + CMC + High Fat Diet) |
9.0 |
% |
|
K2
(Aloxan + CMC + High Fat Diet) |
8.7 |
% |
|
K3 (Aloxan
+ High Fat Diet +
Pioglitazone) |
3.9 |
% |
|
K3 (Aloxan
+ High Fat Diet +
Pioglitazone) |
3.8 |
% |
|
K3 (Aloxan
+ High Fat Diet +
Pioglitazone) |
4.1 |
% |
|
K3 (Aloxan
+ High Fat Diet +
Pioglitazone) |
3.6 |
% |
|
K3 (Aloxan
+ High Fat Diet +
Pioglitazone) |
4.2 |
% |
|
K4 (Aloxan
+ High Fat Diet +
Extract) |
3.2 |
% |
|
K4 (Aloxan
+ High Fat Diet +
Extract) |
3.3 |
% |
|
K4 (Aloxan
+ High Fat Diet +
Extract) |
3.1 |
% |
|
K4 (Aloxan
+ High Fat Diet +
Extract) |
3.5 |
% |
|
K4 (Aloxan
+ High Fat Diet +
Extract) |
3.6 |
% |
|
K5 (Aloxan
+ High Fat Diet +
Pioglitazone + Extract) |
4.3 |
% |
|
K5 (Aloxan
+ High Fat Diet +
Pioglitazone + Extract) |
4.0 |
% |
|
K5 (Aloxan
+ High Fat Diet +
Pioglitazone + Extract) |
3.9 |
% |
|
K5 (Aloxan
+ High Fat Diet +
Pioglitazone + Extract) |
4.1 |
% |
|
K5 (Aloxan
+ High Fat Diet +
Pioglitazone + Extract) |
3.5 |
% |
����������� The HbA1c level was first analyzed
using the Shapiro-Wilk test to check whether the data was distributed normally,
and the results of the normally distributed data were obtained (p > 0.05).
Then the data was analyzed by the ANOVA one-way test. The results of this
analysis are shown in the following table:
Table 1. Normality Test and One Way ANOVA
|
Group Treatment |
Mean �
SD |
Normality Shapiro
Wilk |
One
Way Annova |
|
|
F
Calculate |
P-value |
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|
K-1 |
3.72 � 0.19 |
0,928 |
445,193 |
<
0.001 |
|
K-2 |
8.94 � 2.32 |
0,685 |
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K-3 |
3.92 � 0.23 |
0,899 |
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K-4 |
3.34 � 0.20 |
0,754 |
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K-5 |
3.96 � 0.29 |
0,777 |
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The results of the ANOVA
one-way �test showed a p-value of
< 0.001, which indicates that the combination of snakehead fish extract,
meniran, and temulawak significantly affected the decrease in HbA1c levels in
the serum of male rats of the wistar strain (Rattus norvegicus) after 26
days of treatment. To identify further differences between treatment
groups,� a post hoc test of Tukey LSD �was performed, as seen in the graph below:
b c b a bc
Figure 2. Tukey LSD Post Hoc Chart
The same letter notation showed no
significant difference in the decrease in HbA1c levels between treatments based
on the Tukey LSD post hoc test �at
a significance level of α = 0.05. The K-2 treatment (aloxan + high fat
diet) showed the highest average HbA1c levels compared to other treatments,
while the K-4 treatment (aloxan + high fat diet + extract) had the
lowest average HbA1c levels. This showed that K-4 was the best treatment group
in lowering HbA1c levels in male rats of the Wistar strain (Rattus norvegicus)
compared to other treatments. Therefore, it can be concluded that the
combination of snakehead fish extract, meniran, and temulawak has a significant
effect on reducing HbA1c levels in rats.
Overview of Pancreatic Histopathology in Diabetes Mellitus Model Rats with a High Fat Diet After Treatment
The results of the study showing the effect of giving a
combination of snakehead fish (Channa striata), meniran (Phyllanthus
niruri L.), and temulawak (Curcuma xanthorrhiza) extracts for 26
days can be seen in Figure 3.
Figure
3. Overview of Histopathology of the Rat Pancreas
The histological picture of the
pancreas of each treatment group shows differences, but there is one thing in
common, namely the absence of atrial cell desquamation, as shown in Figure 3.
The picture of pancreatic histology did not change in the normal group (a).
Group 2 (b) (aloksan + CMC + high fat diet) also showed no
abnormalities. Group 3 (c) (aloksan + high fat diet + pioglitazone)
showed Langerhans islet atrophy, cell nucleus picnicosis, as well as fading
cytoplasm on the pancreatic Langerhans islet. Group 4 (d) (aloksan + high
fat diet + extract) showed similar results to group 3 (c), namely
Langerhans islet atrophy, cell nucleus picnicosis, and fading cytoplasm on the
pancreatic Langerhans islet. Group 5 (e) (aloxan + high fat diet +
pioglitazone + extract) showed no abnormalities.
Discussion
The treatment group was given a
high-fat diet for about 26 days, with additional feed in the form of lard 3
grams/200 gramsBB and duck egg yolk 2 grams/200 gramsBB, while group 1
(negative control) did not get any treatment. The treatment results showed that
a high-fat diet increased the weight of rats by 10 grams. The analysis
of the Kruskal-Wallis test showed that there was a significant
difference in the weight gain of male rats of the Wistar strain with a value of
p = 0.016 (< 0.05) between the treatment groups.�
Intraperitoneal
induction of aloxan at a dose of 100 mg/kgBB led to an increase in blood
glucose levels in groups 2, 3, 4 and 5 up to 300 mg/dL. After the increase,
male rats of the Wistar strain were given a combination of snakehead fish
extract, meniran, and temulawak, as well as pioglitazone for approximately 26
days. On the 14th day after the combination of the extract and pioglitazone,
blood glucose levels dropped to 100 mg/dL.�
The Kruskal-Wallis test on day 14 showed a significant difference
in blood glucose levels of male rats of the Wistar strain with a value of p =
0.003 (< 0.05) between treatment groups. The Mann-Whitney test showed
that there was no significant difference in the reduction of blood glucose
levels between the treatment groups (p = 0.05). However, figure 13 shows that
group 5 experienced the largest decrease in blood glucose levels compared to
other groups. This showed that the combination of pioglitazone at a dose of
0.27 mg/200 gramsBB and the combination of extract at a dose of 9.9 mg/200
gramsBB was the most effective in lowering blood glucose levels.
The
combination of snakehead fish extract, meniran, temulawak and pioglitazone for
approximately 26 days had an effect on HbA1c levels in male rats of the Wistar
strain. Based on the ANOVA one-way test, the combination of extract and
pioglitazone significantly reduced HbA1c levels with a p < value of 0.001. The
Tukey LSD post hoc test showed that there was no significant difference in
the reduction in HbA1c levels between the treatment groups (p = 0.005).
However, group 2 had the highest average HbA1c levels compared to other groups.
Meanwhile, group 4 (aloksan + high-fat diet + extract) with a dose of
9.9 mg/200gramBB showed the most optimal decrease in HbA1c levels. In this
study, drug interactions were also found. The interactions that occurred were
in the form of synergistic/additive interactions and antagonists that could
potentially cause adverse drug reactions (Al
Mukminah & Indradi, 2021). In group 5, HbA1c levels decreased by 3.96%, while group 4 decreased
by 3.34%. From these results, it can be concluded that there is an interaction
between the combination of the extract and pioglitazone that affects the
decrease in HbA1c levels in group 5.
Research shows
that therapy with snakehead fish extract at a dose of 300 mg/kgBB and metformin
at a dose of 45 mg/kgBB is better than single therapy with EIG or metformin. In
addition, it was found that meniran leaf extract at a dose of 200 mg/kgBB lowered
blood glucose levels from 200 mg/dL to 90 mg/dL. The study also supported these
findings with a dose of 18 mg/kgBB of temulawak extract could lower blood
glucose levels from 206 mg/dL to 123 mg/dL, while a dose of 20 mg/kgBB lowered
glucose levels to 101 mg/dL from 214 mg/dL. (Nurcahyani, 2022) (Yulizal et al.,
2021) (H. Sari et al.,
2019)
The
combination of snakehead fish extract, meniran, temulawak, and pioglitazone for
26 days also provided a typical histopathological picture of the pancreas,
especially Langerhans Island. In groups 3 and 4, it was shown that there was
atrophy of Langerhans island, pyrosis of cell nucleus, and fading cytoplasm on
Langerhans island pancreas and no desquamation was found. The other treatment
groups did not show any abnormalities.
The study
showed that histopathology in the pancreas of rats with aloxan-induced
hyperglycemia showed that Langerhans Island was atrophied. In addition, it is
also seen that the cell nucleus with psychosis or condensation of the cell
nucleus is darker due to more intense staining. Langerhans Island the pancreas
undergoes desquamation and cytoplasm that appears to fade.� stated that various factors can cause pincosis,
atrophy of the cell nucleus, and changes in the histopathology of the
Langerhans islet pancreas of mice. Oxidative stress, which occurs as a side
effect of pioglitazone, triggers the degeneration of endocrine cells, leading
to picnicnics and atrophy. In addition, the presence of bioactive compounds
such as flavonoids, which act as antioxidants can have different effects. The
histopathological condition of the pancreas may worsen if the dose or
concentration of the extract is not appropriate or if there is a negative
interaction with pioglitazone
(Maharani et al.,
2023) revealed that aloxan does not always cause morphological changes in
pancreatic tissue, such as pincosis or atrophy, although it can trigger necrosis.
Aloxans tend to cause necrosis without causing more subtle changes, such as
atrophy or pincosis.� It is also
suggested that histopathological results are affected by the dose of aloxan and
the method of administration. Lower doses or certain methods of administration,
such as intraperitoneal, may not be strong enough to alter significant
histopathological changes, so the resulting picture differs from those of
higher doses or other methods (Wulandari et al.,
2024)v.�����
Several natural compounds work together to lower blood glucose
levels HbA1c levels and improve the histopathological picture of the pancreas
observed in this study. The amino acids in snakehead fish proteins, such as
arginine and leucine, control blood glucose levels in hyperglycemia. The
antihyperglycemic and anti-inflammatory properties of �(Soniya & Fauziah, 2020) C.xanthorrhiza and
xanthorrhizole extracts make it an effective antidiabetic agent in the
treatment of diabetes mellitus. In addition, meniran contains alkaloids that
function as antipyretic, antidiarrheal, and antidiabetic. Filantin and
hypophyllant compounds that function as hepatoprotectors, as well as
flavonoids, quercetin and nirurin that act as anti-carcinogens, also function
as diuretics (Maulida & Indradi, 2019), (Risnawati et al., 2021).
CONCLUSION
The
study findings indicate that the combination of snakehead fish extract (Channa
striata), meniran (Phyllanthus niruri L.), curcuma (Curcuma xanthorrhiza), and
pioglitazone is an effective method for reducing blood glucose levels in male
Wistar rats. Furthermore, administration of a combination of snakehead fish
extract, meniran, and curcuma for a period of 26 days demonstrated a notable
reduction in HbA1c levels, indicative of enhanced glycemic control. However,
histopathological analysis demonstrated that neither pioglitazone nor the
combined treatment resulted in an improvement in the condition of the islets of
Langerhans. This lack of improvement may be attributed to various contributing
factors, including oxidative stress resulting from pioglitazone's side effects,
potential inaccuracies in extract dosage or concentration, and the impact of
alloxan induction, including its specific dosage and administration methods.
These findings highlight the combination's potential efficacy in managing blood
glucose and HbA1c levels, but underscore the need for further investigation
into optimal dosing and mitigation of associated oxidative stress to support
pancreatic health.
Further research is needed to understand the effects
of combining snakehead fish extract, meniran, and temulawak with pioglitazone,
on glycemic status and pancreatic histopathology in male rats. Furthermore,
comprehensive studies from a biomolecular perspective are essential to enhance
insights into the mechanisms and potential benefits of these extracts in the
treatment of diabetes mellitus. Such research could illuminate their
therapeutic roles and guide more effective applications in the management of diabetes.
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