Effectiveness of Fig Leaf Extract (Ficus Carica L.) in Lowering Blood Glucose in Mice (Mus Musculus)
Nur Aisyaha,1, Sisilia TR Dewia,2, Jumaina,3
aDepartment
of Pharmacy, Poltekkes Kemenkes
Makassar,
Indonesia
[email protected]1, [email protected]2
ABSTRACT
Fig leaves (Ficus carica L.) is a plant that has antidiabetic properties,
because fig leaves contain alkaloid compounds, flavonoids, tannins,
polyphenols, saponins, terpenoids, and steroids. This study aims to determine
the extract of fig leaves (Ficus carica L.)
in lowering blood glucose in mice (Mus musculus). Extraction is carried out by
maceration with 96% ethanol solvent, then evaporated using Rotavapor. This
study used alloxane as a diabetes inducer, the dose of fig leaf extract tested
was 5%, 10% and 15% given orally. Observations were made on day 3 and day 7. As
a positive control used Glibenclamide and Na. CMC 1% w/v as a negative control.
The results showed an average percent reduction in blood glucose levels for
negative controls by 18.6%, for 5% extracts by 29.1%, for 10% extracts by
46.6%, for 15% extracts by 83.9% and for positive controls by 45.8%. The
results of research have been obtained that the administration of Fig Leaf
extract (Ficus carica L.)
is proven to reduce blood glucose levels in mice (Mus musculus) induced by
alloxane at concentrations of 5%, 10%, 15%. In the One-way ANOVA test, it shows
a significant value (p < 0.05) showing a significant difference in each
contract. It can be concluded that fig leaves can be a potential alternative
treatment for diabetes mellitus with the right concentration can be an
effective and natural treatment option.
Keywords: alloxan, diabetes, ficus carica l.
Correspondent: Nur Aisyah
Email: [email protected]�
INTRODUCTION
Diabetes
mellitus is the most deadly disease globally (Lampe et al., 2020).
This disease causes complications that lead to death. Diabetes mellitus
is a disease that cannot be infected. Diabetes is divided into several types:
Type I and Type II. Every year, there is always a significant increase (Kurniawaty &
Lestari, 2016).
Quoted
from the Data and Information Center of the Indonesian Ministry of Health
(InfoDATIN) states that Diabetes does not only cause premature death worldwide.
This disease is also a significant cause of blindness. Heart disease and kidney
failure. The International Diabetes
Federation (IDF) estimates that at least 463 million people aged 20-79 years in
the world have Diabetes in 2019 or the equivalent of a prevalence rate of 9.3%
of the total population at the same age. Based on gender, IDF estimates that
the prevalence of Diabetes in 2019 is 9% for women and 9.65% for men. The
prevalence of Diabetes is estimated to increase with the increasing age of the
population to 19.9% or 111.2 million people aged 65-79 years. The number is
predicted to continue to increase until it reaches 578 million in 2030 and 700
million in 2045.
The 2018 Riskedas results show that the prevalence of Diabetes mellitus
in Indonesia based on a doctor's diagnosis at the age of ≥15 years on the
2013 Riskesdas results is 1.5% (Karyadi, 2022). However, the prevalence of Diabetes mellitus,
according to blood sugar examination results, increased from 6.9% in 2013 to
8.5 in 2018. This figure indicates that only around 25% of diabetics know that
they have Diabetes (Nova et al., 2020).
The
World Health Organization (2018) estimates that around 422 million adults aged
over 18 years have been living with Diabetes since 2014. The most significant
number are estimated to come from Southeast Asia and the West Pacific, 96
million and 131 million, respectively.
In
the world of medicine, this disease can be treated with various kinds of drugs,
such as taking medicines made from herbal ingredients or chemical drugs.
Treatment efforts to control blood sugar levels and prevent complications in
people with diabetes mellitus include using hypoglycemic drugs derived from
plants. Ara leaves are plants used in traditional medicine, and fig leaves have
many benefits (Adawiyah, 2018).
One
of the plants that have antidiabetic activity is the fig or fig fruit, but what
is often used from this plant is the leaves to lower blood glucose in the body (WU et al., 2022). Fig
leaves are a good source of vitamins A, B1 and B2. Ara leaves also contain
calcium, phosphorus, manganese, iron, potassium and
sodium. Fig leaves have many health benefits (Maximum, 2020). �
Based
on research conducted by (Nafiandary, 2019), Fig
leaves (Ficus carica L) contain phytochemical compounds belonging to the
alkaloid, phenolic, flavonoid and triterpenoid groups, which can treat various
health problems such as cancer, diabetes mellitus, vascular and nerve diseases.
Plants
are often used directly by the people of Bone, especially in the Lapri area,
because they are alternative medicines for the treatment of Diabetes, which
have fewer side effects than chemical drugs (Adhikari et al.,
2023). Even though fig leaves have been used empirically, if they have not been
accepted in the medical world, then fig leaves will still be familiar with
people's herbal medicines. Fig leaves without proof will not develop into
herbal products that people trust. Researchers are interested in researching
fig leaves (Ficus carica L) for their effectiveness. However, further
research is needed because many plants have unknown toxicity levels. In
addition to Diabetes medication, fig leaves also have properties such as
preventing heart disease, anti-cancer, and heart disease (Lestario, 2018).
The purpose of this study was to see the ethanol extract of fig leaves (Ficus
carica L) has an effect in lowering blood glucose in mice (Mus musculus).
The benefit of this study is to provide information to the public about the use
of Fig Leaves in lowering blood glucose and as a reference for future research.
METHODS
This research was conducted at the Pharmacology Laboratory and Chemistry
Laboratory of the Pharmacy Department of the Health Ministry's Makassar
Polytechnic. The time for this research to be carried out is between August
2021- April 2022. This type of research is experimental research conducted to
test fig leaf extract (Ficus carica L.) obtained from the district. Bone
in the Lapri Section to test the effectiveness of reducing blood glucose in
mice (Mus musculus). �The materials used
in this study were fig leaves (Ficus
carica L.) and mice (Mus musculus), 96% ethanol,
glibenclamide tablets, ammonia, chloroform, H2SO4, Mayer reagent, Wagner
reagent, dragendroff reagent. Concentrated HCl, Mg Powder, FeCl3,
Acetic Acid, Na-CMC, and alloxan. The tools used in
this study were GlucoDr AGM-2100, glucose strips, syringe, analytical balance (Aicis), coarse scale (Ohaus), stir bar, porcelain cup,
measuring cup, maceration vessel, simplicial oven, rotary evaporator, tube�s reaction, Erlenmeyer 100 ml, mortar, scissors,
and alcohol swab. �The experimental animals used in this study
were mice (Mus musculus) healthy males (@ 30 g) totaling 15 heads who were
monitored for one week and gave feed and water ad libitum and were divided into
2 groups, namely groups 1 and 5 were control groups (6 mice) which were divided
into two groups, namely the positive control group and the negative control
group without alloxane inducers. While group 2,3,4 is the treatment group.
Furthermore, mice were induced using alloxane at a dose of 3.36 mg / 20 g / 0.5
ml / 20 g administered intraperitonial and fed and drank for 3 days.
Initial
measurement of blood glucose levels was carried out on day 3 and day 7 after
administration of the extract. blood collection was carried out through the
tail of mice by first satisfying mice for 8 hours. Mice are declared diabetic
if their fasting blood glucose level is more than 150mg/dL. After the treatment
was carried out, the mice were checked again using the GlucoDr AGM-2100
glucometer with previously satisfied for 8 hours. Furthermore, data on glucose
levels from male mice (Muc musculus) were compiled in tables. The percentage
results that have been obtained, then statistical analysis is carried out using
the SPSS version 22 test. To see the normality value with the Shapiro-Wilk
method, the result shows that the significant value (p > 0.05) is a normal
distributed value. The test looks at the homogeneity value with the Test of
Homoginrtiy of Variences method, the results show Sig values of 0.56 > 0.05
which means homogeneous data. The variable inhibitory power test data is
homogeny and can be continued for the ANOVA One way test. One way ANOVA test
results obtained values of 0.000 < 0.05 which means there are significant
differences between each treatment group. And then continued with further
tests, namely Post Hoc with the Tukey method obtained a significant value (p
< 0.05) showing a difference between each concentration. Each observation
was analyzed using analysis of variant (ANOVA).
RESULTS AND DISCUSSION
Results Leaf Extraction Fig (Ficus carica L.)
This time, the
sample was Ara Leaf (Ficus carica L), taken from the district.
Bone, Kususnya in parts area Capri Which picked direct from the tree.
Table 1.
Heavy Extract Leaf Fig (Ficus carica L.)
|
Sample |
Type Solvent |
Heavy Sample dry |
Heavy Extract |
yield |
Literature |
|
Leaf Fig (Ficus carica L.) |
Ethanol 96 % |
165 grams |
91.7 grams |
55.57% |
< 5.9% (Senduk et al., 2020) |
Results Test Compound
Phytochemicals Leaf Extract Fig (Ficus carica
L.)
Table 2.
Results Observation Screening Phytochemicals Extract
Leaf Fig
|
Type compound |
Change colour |
Results |
References |
|
Alkaloids |
Mayer = precipitate white Wagner = precipitate chocolate |
(+) |
Mayer = precipitate white Wagner = precipitate chocolate |
|
Flavonoids |
precipitate Green |
(+) |
Green |
|
tannins |
Green black |
(+) |
Green Black |
|
Polyphenols |
Black |
(+) |
Black |
|
Saponins |
formed froth |
(+) |
formed froth |
|
terpenoids |
Purplish |
(+) |
Purple |
|
Steroids |
Own black ring |
(-) |
Blue |
Results of
Measuring Blood Glucose Levels in Mice Which�� induced Alloxan
Table 3. Results of Measuring Blood Glucose Levels in
Induced Mice Alloxan
|
Group |
Replication |
Heavy Mice (g) |
Sugar Blood
Beginning (mg/dL) |
After Induction (mg/dL) |
Blood Glucose Levels After Giving (mg/dL) |
|
|
Day The
3rd |
7th day |
|||||
|
Na.CMC 1% (Control
-) |
1 |
29.4 g |
75 |
117 |
113 |
110 |
|
2 |
21.9 g |
153 |
193 |
189 |
183 |
|
|
3 |
27.2 g |
84 |
112 |
110 |
108 |
|
|
Amount |
|
312 |
422 |
412 |
401 |
|
|
Average |
|
104 |
140.6 |
137,3 |
133.6 |
|
|
Extract 5% |
1 |
27.2 g |
129 |
152 |
149 |
147 |
|
2 |
26.5 g |
134 |
200 |
196 |
172 |
|
|
3 |
27.0 g |
175 |
182 |
179 |
172 |
|
|
Amount |
|
438 |
534 |
521 |
491 |
|
|
Average |
|
146 |
178 |
173.6 |
163.6 |
|
|
Extract 10% |
1 |
30 g |
119 |
247 |
191 |
185 |
|
2 |
26.5 g |
130 |
254 |
210 |
198 |
|
|
3 |
28.1 g |
125 |
237 |
220 |
185 |
|
|
Amount |
|
374 |
738 |
621 |
568 |
|
|
Average |
|
124.6 |
246 |
207 |
189.3 |
|
|
Extract 15% |
1 |
31.2 g |
125 |
174 |
161 |
133 |
|
2 |
30.7 g |
97 |
277 |
259 |
128 |
|
|
3 |
29.8 g |
130 |
189 |
143 |
120 |
|
|
Amount |
|
352 |
640 |
563 |
381 |
|
|
Average |
|
117,3 |
213,3 |
187.6 |
127 |
|
|
Glibenclamide (Control +) |
1 |
27.6 g |
165 |
200 |
192 |
185 |
|
2 |
19.3 g |
125 |
260 |
245 |
195 |
|
|
3 |
26.8 g |
80 |
230 |
200 |
160 |
|
|
Amount |
|
370 |
690 |
637 |
364 |
|
|
Average |
|
123.3 |
230 |
212,3 |
121.3 |
|
Average Blood Glucose Levels
of Mice at the Beginning, After Induction and Decrease in Blood Glucose
Figure 1. Mean Blood Glucose
Levels in Early Mice,
After Induction and Decrease
in Blood Glucose
Percentage of Decreased Blood
Glucose Levels of Mice (Mus musculus) as Indicated by Alloxan
Table 4. Percentage of
Decreased Blood Glucose Levels in Alloxan-Induced Mice
|
Replication |
Percentage Effect Decline Rate Glucose Blood
Mice |
||||
|
Control Negative (Na-CMC 1%) |
Extract Fig leaf ( Ficus carica
L.) 5% |
Extract Fig leaf ( Ficus carica L.) 10% |
Extract Fig leaf ( Ficus carica
L.) 15% |
Positive Control (Glibenkamid) |
|
|
1 |
16.6% |
21.7% |
48.4% |
83.6% |
42.8% |
|
2 |
25% |
42.4% |
45.1% |
82.7% |
48.1% |
|
3 |
14.2% |
23.3% |
46.4% |
85.5% |
46.6% |
|
Average |
18.6% |
29.1% |
46.6% |
83.9 |
45.8% |
STDEV Results of Blood Glucose Levels in Mice (Mus Musculus)
Figure 2. The average percentage of blood glucose
levels
Based on the research results, samples of fig leaves (Ficus carica L.)
are soaked using 96% ethanol. 96% ethanol was chosen as the solvent in this
extraction because, according to research conducted by (Nurzahra et al., 2022), the best treatment test
results were obtained in the 96% ethanol solvent type treatment. 96% ethanol is
a polar compound that quickly evaporates, so it is good to use as an extracting
solvent (Kurniawati, 2017). The weight of the dry sample
is 165 grams, while the weight of the viscous extract is 91.7 grams, and the
yield is 55.57%. Yield is the ratio of dry weight to the number of raw
materials. The yield value is related to the amount of bioactive content contained.
The higher the yield, the higher the content of substances attracted to raw
material. This calculation is carried out to determine the percentage of the
amount of material remaining as a result of the extraction process and to
determine the effectiveness of the resulting process (Farhan et al., 2022).
Based on the results of the phytochemical
screening test of Fig Leaf Extract (Ficus carica L.), Alkaloid testing
using Mayer's reagent (White precipitate) on Wagner's reagent (brown), it can
be concluded that (alkaloid positive) (Meziant et al., 2021). Flavonoid testing showed
results (green precipitate) on adding Mg metal and concentrated HCl, concluded
(flavonoid positive). The tannin test showed results (blackish Green) after
adding FeCl3 solution and was concluded (tannin positive). Polyphenol testing
shows (black colour) concluded (polyphenol positive). The saponin test shows
(favourable for the formation of foam) after boiling for 2-3 minutes and
cooling, it is concluded (positive for saponins). The terpenoid test showed
(purple colour) after adding glacial acetic acid, and it was concluded
(terpenoid positive). Steroid testing showed results (formed black rings), and
it can be concluded that fig leaves do not contain steroid compounds (Meziant et al., 2021). Therefore, fig leaves
contain alkaloids, tannins, flavonoids, polyphenols, saponins and terpenoids.
Meanwhile, steroids are not contained in fig leaves (Simaremare, 2014).
In this study, the test animals used were male
mice weighing 20-30 g, first adapted to the surrounding environment. Mice were
chosen as test animals because they have well-characterized anatomical and
physiological characteristics; besides that, they are easy to obtain, easy to
handle, inexpensive, and considering that the volume of blood needed to measure
blood sugar levels is only tiny, it will be more effective to use mice than
other animals. Male mice were chosen because they have a more stable hormone
system than female mice. After all, the hormone estrogen in female mice can
affect blood glucose levels in the body. Male mice aged 2-3 months are young
adult mice with optimal physiological conditions (Khairiyani, 2018). Mice were divided into five
groups. Group 1 is negative control (Na-CMC 1%), group 2 is 5% extract, group 3
is 10% extract, group 4 is 15% extract and group 5 namely positive control
(glibenclamide), where glibenclamide is an antihyperglycemic drug, sulfonylurea
group with a mechanism stimulates insulin secretion.
Table 3 shows the results of observations that have been treated in the
five groups of mice. The results of observations of initial blood glucose
levels ranged from 75-165 mg/dL before being induced by alloxan. Blood glucose
levels after alloxan induction ranged from 117-277 mg/dL. Mice with fasting
blood glucose levels ≥ 150 mg/dL are declared hyperglycemia and can be
used for further studies (Rachmansyah et al., 2020). This shows that alloxan has
damaged the pancreatic beta cells of mice. However, when alloxan was given,
glucose was not added so that these mice could experience hypoglycemia, so
these mice did not experience a significant increase in blood glucose. Based on
research conducted by (Kumalasari et al., 2019), giving 5% glucose after
alloxan-induced mice aim to prevent the hypoglycemia phase, which can cause
seizures and can be fatal.
The bar chart shows that the blood sugar level before and
after induction increased, and after being given treatment, i.e. administration of fig leaf extract,
decreased significantly. The inducer
used is alloxan because alloxan is fast and can
reach the pancreas; its action is preceded by rapid uptake by Langerhans β cells. The
formation of reactive oxygen is a primary factor in damage to cells
(Surbakti & Khairun, 2015). Giving There
was a decrease in the extract. The Ara Leaf extract is suspected of
containing. Phytochemicals, namely alkaloids, flavonoids,
tannins, polyphenols, saponins, and terpenoids. This is where flavonoid compounds can lower blood glucose levels with the
ability as substance antioxidant. Flavonoids are characteristic protective to damage cell β as producer insulin
and can increase
insulin sensitivity (Liwu et al., 2019). Alkaloids in Leaves Ara (Ficus carica L.)
have the potential as anticancer and antidiabetic. For Tannins, polyphenols, saponins, terpenoids and steroids
contained in Ara Leaf extract can
also increase glycolysis which can help lower glucose blood in the body.
However, compared to compound flavonoids, it can be concluded that
the content of flavonoids is relatively high in the treatment of antidiabetic.
Generally, the contraction extract,
Which is most effective
in lower blood glucose in mice, is a 15%
concentration. This matter because
fig leaf extract (Ficus carica L)
contains compounds active that existing content flavonoids Which own antidiabetic ability Which Work in the
process of regeneration from the
cell beta pancreas And increase insulin secretion.
CONCLUSION
Based on the results of research that has been done,
it can be concluded that fig leaves
with a concentration of 5%, 10% and 15% have an effectiveness decline
in glucose levels blood in mice Which induces
alloxan. Ara leaves can be a potential
alternative treatment for diabetes mellitus. In treating Diabetes mellitus,
especially type 2, lowering blood glucose levels is crucial to avoid more
severe complications. Using Ara leaves with the right concentration can be an
effective and natural treatment option.
REFERENCES
Adawiyah,
R. (2018). Antifunction test of earring leaf extract (Acalypha indica Linn)
against Trichophyton rubrum and Candida albicans with various solvents.
Maulana Malik Ibrahim State Islamic University.
Adhikari,
YP, Bhandari, P., Adhikari, DM, & Kunwar, RM (2023). Chapter 18 - Ficus species (Ficus auriculata Lour., Ficus
benghalensis L., Ficus carica L., Ficus religiosa L., Ficus semicordata Buch.
Ham. ex Sm). (T. Belwal, I. Bhatt, & HBT-HF and B. Devkota
(eds.); pp. 171�182). Academic Press.
https://doi.org/10.1016/B978-0-323-85591-4.00030-1
Farhan,
MI, Chusniasih, D., & Marcellia, S. (2022). Antibacterial Activity Test of
Tin Leaf Extract (Ficus Carica L.) Against Escherichia coli and Staphylococcus
aureus. PHARMACON , 11 (1),
1328�1334. https://doi.org/10.35799/pha.11.2022.39145
Karyadi,
D. (2022). The Relationship Between Mindfulness And Stress In Individuals
With Diabetes Mellitus. University of Mercu Buana Yogyakarta.
Khairiyani,
A. (2018). Test of the Effect of Reducing Blood Glucose Levels of Ethanol
Extract of Pandan Wangi Leaves (Pandanus amaryllifolius Roxb.) Against
Alloxan-Induced Male Mice. The University of Northern Sumatra.
Kumalasari,
E., Susanto, Y., Rahmi, MY, & Febrianty, DR (2019). The Effect of Ramania
Leaf Ethanol Extract (Bouea macrophylla Griffith) on Decreased Blood Sugar
Levels of White Mice (Mus musculus) induced by alloxan. JCPS (Journal of
Current Pharmaceutical Sciences), 2 (2), 173�179.
Kurniawati,
A. (2017). The Effect of Solvent Type on the Extraction Process of Rose Flowers
Using the Maceration Method as a Perfume Fragrance. Journal of Student
Creativity, 2 (2),
74�83. https://doi.org/10.15294/jcs.v2i2.14587
Kurniawaty,
E., & Lestari, EE (2016). Test the effectiveness of starfruit leaves
(Averrhoa bilimbi L.) as a treatment for diabetes mellitus. Majority Journal,
5 (2), 32�36.
Lampe,
M., Waris, L., & Rajab, M. (2020). The influence of socio-economic gender's
roles towards Diabetes symptoms in South Sulawesi fishermen community. Enfermer�a
Cl�nica , 30 , 140�143. https://doi.org/10.1016/j.enfcli.2019.07.065
Lestario,
LN (2018). ANTHOCYANIN: Chemical Properties, Its Role in Health, and Its
Prospects as a Food Coloring. UGMPRESS.
Liwu,
AN, Lidia, K., & Amat, ALS (2019). Effect of Ethanol Extract of South
African Leaves (Vernonia Amygdalina Delile) on Blood Glucose Levels of Alloxan-Induced
White Rats (Rattus Norvegicus). Sandalwood Medical Journal (CMJ), 7 (2), 299�307. https://doi.org/10.35508/cmj.v7i2.1803
Makmun,
A. (2020). Some of the Benefits of Figs (Ficus Carica) From Anticonvulsants,
Anti Allergic, Anti Inflammatory, Antihyperglycemic, Antitumor, Anti Cancer to
Liver Therapy. Journal of Medicine, 9 (3), 184�201.
Meziant,
L., Bachir-bey, M., Bensouici, C., Saci, F., Boutiche, M., & Louaileche, H.
(2021). Assessment of inhibitory properties of flavonoid-rich fig (Ficus carica
L.) peel extracts against tyrosinase, α-glucosidase, urease and
cholinesterase enzymes, and relationship with antioxidant activity. European
Journal of Integrative Medicine, 43, 101272.
https://doi.org/https://doi.org/10.1016/j.eujim.2020.101272
Nafiandary,
M. (2019). Effect of fig fruit extract (Ficus carica l.) as an antioxidant
on the epithelial thickness and lumen diameter of the proximal tubule of the
kidney of female mice exposed to rhodamine B. Maulana Malik Ibrahim State
Islamic University.
Nova, E., Redondo-Useros, N., Mart�nez-Garc�a, RM,
G�mez-Mart�nez, S., D�az-Prieto, LE, & Marcos, A. (2020). The
potential of Moringa oleifera to improve glucose control for the prevention of
Diabetes and related metabolic alterations: a systematic review of animal and
human studies. Nutrients, 12 (7), 2050. https://doi.org/10.3390/nu12072050
Nurzahra,
A., Mulqie, L., & Hazar, S. (2022). Determination of Total Ash Content and
Specific Weight of Figs (Ficus carica L.). Bandung Conference Series:
Pharmacy , 2 (2),
891�899. https://doi.org/10.29313/bcsp.v2i2.4677
Rachmansyah,
MS, Junaidi, E., & Effendi, E. (2020). Reduction of Blood Glucose Levels by
Apple Vinegar in Mice Induced by Dexamethasone. Journal of Agromedicine and
Medical Sciences, 6 (3),
143�148. https://doi.org/10.19184/ams.v6i3.11481
Senduk,
TW, Montolalu, L., & Dotulong, V. (2020). The yield of boiled water extract
of sonneratia alba old mangrove leaves. Fisheries and Tropical Marine, 11 (1), 9�15.
Simaremare,
ES (2014). Phytochemical screening of ethanol extract of itching leaves
(Laportea document (Roxb.) Wedd). PHARMACY: Indonesian Pharmaceutical
Journal (Pharmaceutical Journal of Indonesia), 11 (1). 10.30595/pji.v11i1.855
Surbakti, ESB, & Khairun, NB (2015). Tomato
(Lycopersicum esculentum Mill.) As anti-ageing Skin. The
majority, 5 (3),
73�78.
WU,
L., GAO, Y., SU, Y., LI, J., REN, W.-C., WANG, Q.-H., & KUANG, H.-X.
(2022). Probiotics with anti-type two diabetes mellitus properties: targets of
polysaccharides from traditional Chinese medicine. Chinese Journal of
Natural Medicines, 20 (9),
641�655. https://doi.org/10.1016/S1875-5364(22)60210-3