Evaluation of Antidiabetic Potential Of

International Journal of Science and Research (IJSR)
ISSN (Online): 2319-7064

Index Copernicus Value (2015): 78.96 | Impact Factor (2015): 6.391
Evaluation of Antidiabetic Potential of

A Novel Formulation from Prophetic Medicine in
Streptozotocin-Induced Diabetic Balb/C Mice
Eman Elm .E. Elaswed1, Moklesur Rahman Sarker2
1
Faculty of Sciences, Lincoln University College Malaysia Mayang Jaya, 47301, Petaling Jaya, Selangor Darul Ehsan, Malaysia
2
Faculty of Pharmacy, Lincoln University College Malaysia Mayang Jaya, 47301, Petaling Jaya, Selangor Darul Ehsan, Malaysia
Abstract: Diabetes is a common, chronic and expensive disease that is threatening the health of generations of people all around the
world. Diabetes mellitus is a disorder that affects the body’s ability to make or use insulin and it remains an incurable disorder in spite of
intense research. Prophetic Medicine is the total authentic Hadith narrated by the Prophet (peace be upon him), which carries incredible
values for the administration of diabetes. The present study aimed to develop a new formulation from Prophetic Medicine and to
evaluate its therapeutic potential for the effective use in diabetes for avoiding major adverse effects in animal model. Towards this aim,
diabetes was induced to Balb/c mice with the injection of streptozotocin (160 mg/kg). The mice were treated with the formulation of
Prophetic medicine (PMFD) at the doses of 200, 400, 600 mg/kg and metformin at dose of 200 mg/kg orally. The glucose levels were
measured on first day for 3 hours, second and third day for 2 hours and 2 weeks before and after treatment with the PMFD.
Non-diabetic mice were used as a negative control and diabetic mice without treatment was used as a positive control .The formulation
(PMFD) at the dose of 200, 400 and 600 mg/kg shows significant reduction in post-prandial plasma glucose level and body weight,
whereas metformin of 200 kg/mg shows significant reduction of glucose level on second and third day.Thus, the present formulation of
Prophetic medicine can be recommended for the prevention and treatment of diabetes. This data can support for biomedical research
efforts that can lead to prevention, treatment and possibly even cures for diabetes.
Keywords: Diabetes Multiuse, Prophetic medicine, Plasma Glucose level, Balb/c mice, Body Weight Streptozotocin , Metformin
1. Introduction The aim of the present study is to investigate or evaluate the

therapeutic potential of PMFD for management and treated
Diabetes mellitus (DM) is a clinical syndrome associated streptozotocin (STZ)-induced diabetic mice with avoiding
with deficiency of insulin secretion or action. It is major adverse effects. This study hypothesize that diabetic
considered one of the largest emerging threats to health in mice treated with PMFD will significantly reduce fasting
the 21st century. Insulin secretion is subject to control by plasma glucose FPG and post-prandial blood glucose PPBG
nutrients, hormonal, neural, and pharmacological factors. levels. The result from this study might be significantly
Among these glucose is the most important regulator of contribute for the management of plasma blood glucose of
insulin secretion machinery (Mosley et al., 2013). It is STZ- induced diabetic Balb/c mice or minimizing the
estimated that there will be 380 million persons with DM in side-effects of diabetic medication. Thus, it can be a clue of
2025 (Atkins & Zimmet, 2010). Despite considerable new drug for the effective management of diabetes
progress in the treatment of diabetes by hypoglycemic improving the quality of life for diabetic patients.
agents, search for newer drugs continues because the
existing synthetic drugs of diabetes are associate with the 2. Objectives
increased risk of many complications, such as
cardiovascular disease, cancer, renal impairment (Cassidy et The aim of the current research is to develop a new
al., 1999). formulation from Prophetic medicines and to evaluate its
therapeutic potential for the effective management of
Many prophetic medicine plants have been used for the plasma glucose STZ- induced diabetic Balb/c mice with
treatment of diabetes mellitus. Out of these only a few have avoiding major adverse- effects in animal model.
been evaluated as per modern system of medicine. From
many such plants, only extracts have been prepared and 3. Methodology
their usefulness evaluated in experimental diabetes in
animals. Unfortunately, there is a lack of scientific 3.1 Materials
investigations to discover and establish the effectiveness of
Prophetic medicines for the treatment of different diseases. 3.1.1. Prophetic materials used in PMFD
Towards, achieving this goal present study was performed. The Essential Prophetic Medicine (PM) materials were
In the current study, streptozotocin (STZ)-induced diabetic selected based on literature survey and their use in
mice were treated by Prophetic medicine formula for traditional medicines. These materials were purchased from
Diabetic (PMFD) and subjected to management of blood Arabian shops and Malaysian markets as well. The list of
glucose level and body weight. the prophetic materials used in the study are listed in Table
1.
Volume 6 Issue 3, March 2017
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Licensed Under Creative Commons Attribution CC BY
Paper ID: ART20171623 DOI: 10.21275/ART20171623 857
Table 1:List of selected Prophetic medicine materials with their supplier
3.1.2.Chemicals cinnamon powder to 500 ml of 80 % ethanol and by shaking

The chemicals used in this study are listed in Table 2 the flask twice a day to mix the material with each other.
together with their source of manufacturer. Alcohol was changed every two days for two weeks. At last
day of extraction, all cinnamon extract was filtered using
Table 2: List of chemicals and reagents used in the study filter paper. Finally, extracted cinnamon was dried by using
No Chemicals and Reagent Source heating mantle at 40 0C temperature.
1 Ethanol Dhe tech enterprise Taman
Mayang, PJ Malaysia 3.2.2.2 Extraction of black seed
2 Citric acid R &N Marketing, Essex, U.K. Black seed powder of 200 g was added to 500 ml of 80 %
3 Sodium citrate Alpha Chemika, India. ethanol and shaking the flask twice a day to mix the
4 Streptozotocin Sigma Aldrich, U.S.A. material with each other and alcohol changed every two
5 Metformin Santa Crutz, German. days for two weeks to get the extraction. At last day of
extraction, all black seed extract was filtered using filter
3.1.3 Experimental Balb/c mice paper. Finally, the extracted black seed were dried by using
Total 18 male Balb/c mice were used for the experimental, heating mantle at 40 0C temperature.
which were purchased from “Pets Wonderland Paradigm
Mall” Selangor Malaysia (Their source: Envigo, USA). The 3.2.2.3 Extraction of barley
approximate weight of each mouse was 18–30 g and the age Barley powder of 300 g was added to 700 ml of 80%
was in between 7-9 weeks. All mice were kept under ethanol and the flask was shaked twice a day to mix the
standard conditions at animal house of Lincoln University material with each other and alcohol changed every two
college. Balb/c mice were fed with normal diet for 10 days days for two weeks. At last day of extraction, all barley
before proceed in the experiments. extract was filtered using filter paper. Finally, extraction of
barley was dried by using heating mantle at 400C
3.2 Methods temperature.
3.2.1 Collection of materials used for the formulation of 3.2.2.4 Extraction of olive leaves
antidiabetic of Prophetic medicine. Olive leaves powder of 200 g was added to 500 ml of 80%
The materials that have been used in this study are ethanol and shake the flask twice a day to mix the material
Cinnamon (Cinnamomum zeylanicum), Ginger (Zingiber with each other and alcohol changed every two days for two
officinale), Black seed (Nigella sativa), Olive leaves (Olea weeks. At last day of extraction, all olive leaves extract was
European), Barley (Hordeum vulgare L), Olive oil, Apple filtered using filter paper. Finally, extraction was dried by
cider vinegar and Pure Honey. Most of materials were dried used heating mantle at 400C temperature.
and crushed into powder and measured, whereas ginger was
washed, cut into small pieces and dried under sun light for 3.2.2.5 Extraction of ginger
one week then crushed into powder and measured. Ginger powder of 150 g was added to 400 ml of 80 %
ethanol and shake the flask twice a day to mix the material
3.2.2 Extraction of materials used in the formulation of with each other and alcohol changed every two days for two
antidibetic Prophetic medicine weeks. At last day of extraction, all ginger extract was
The Extraction of materials performed by grinding and filtered using filter paper. Finally, extracted ginger was dried
drying of each material into the powder and proceeds with by using heating mantle at 40 0C temperature. All extracted
extraction. All flasks containing the mixture of material samples of PMFD kept in fridge until it prepared the
powders in ethanol were kept in a dark place for all formulation.
extraction processing.
3.2.3 Preparation of formulated Prophetic medicine
3.2.2.1 Extraction of cinnamon After three weeks of processing PMFD materials, different
The extraction of Cinnamon was performed by adding 300 g quantity of each PM extracted material was measured
separately by using electrical balance. Cinnamon (2 g),
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Black seed (2 g), Barley (1 g), Olive leaves (2 g), Ginger (3 with PMFD. Blood glucose levels was measured also for
g), Olive oil (5ml), Apple cider vinegar (1 ml), and Pure non-diabetic, diabetic and metformin group. For all the 6
Honey (15 ml) were mixed together in a beaker. Finally, groups blood glucose level were measured on first day
mixer of formulation was kept in the fridge until further use. before and after 3 hours, second and third days before and
after 2 hours and 2 weeks before and after treatment in
3.2.4 Induction of diabetes in Balb/c mice by administrated mice.
streptozotocin
Diabetes was induced in Balb/c mice by a single 3.2.7 Measurement of body weight
intraperitoneal injection of 160 mg/kg weight of 0.1 M Body weight of mice was measured on the first and last
streptozotocin. STZ was prepared in 0.1 M citrate buffer by days of the experiment (before and after treatment with
dissolving 2.1 g of citric acid and 2.94 g of sodium citrate in PMFD) by using Electric balance.
100 ml of distilled water. The pH was adjusted to 4.5 by
proper addition of concentrated NaOH/HCL using a 3.3 Data analysis
calibrated pH meter (Ballester et al., 2004). After 48 hours
of induction of diabetes, One touch Glucometer and Statistical Analysis was performed by using IBM SPSS 22
compatible glucometer strips were used for the version software. The results for blood glucose level were
determination of blood glucose levels in over-night fasted presented as mean ± S.D (standard deviation). One way
mice. analysis of variance (ANOVA) test was used in this study.
Differences were considered statistically significant at P <
Blood samples were collected from dorsal vein of the tail of 0.05.
conscious mice. Diabetes was allowed to stabilize for 5 days
before the commencement of intervention. Thereafter, the 4. Results
blood glucose level of all animals in each experimental
group was assessed. Animals were checked for clinical signs The current research was done in lab environment at
of drug toxicity such as tremors, diarrhea, weakness, weight Lincoln University. Total 18 mice were used in this project,
loss and death. which were equally divided into 6 different groups that is 3
mice each group.
3.2.5 Treatment of mice with prophetic medicine
formula (PMFD) Diabetes was induced in Balb/c mice with the i.p.
After preparation of final prophetic medicine formula administration of streptozotocin. The administration of
(PMFD) then, all Balb/c mice were divided into 6 prophetic medicine formula (PMFD) given to group 3, 4 and
experimental groups and different doses of PMFD was 5 groups as 200, 400 and 600 mg/kg dose. Group 6 diabetic
given as listed below. mice were treated with metformin at dose of 200 mg/kg.
Non-diabetic Mice (group-1) used as a negative control and
Group 1: Non-diabetic Mice (negative control-1) diabetic mice without treatment was used as a positive
Group 2: Diabetic Mice without treatment (positive control- control (group-2). Blood glucose levels of each mice of all
2) the groups were measured on first day before and after
Group 3: Diabetic mice treated with PMFD (200 mg/kg) treatment of 3 hours, second and third day before and after
Group 4: Diabetic mice treated with PMFD (400 mg/kg) treatment of 2 hours, and after 2 weeks. Body weight of
Group 5: Diabetic mice treated with PMFD (600 mg/kg) mice were measured on 2nd week before and after treatment
Group 6: Diabetic mice treated with metformin (standard. with PMFD.
Antidiabetic drug) at dose of 200 mg/kg.
4.1 Result of PMFD on blood glucose levels in
Nondiabetic (group-1) and diabetic (group-2) mice was STZ-induced Balb/c mice 3 hours after treatment of
administrated with distilled water daily and group 3, 4 & 5 diabetic mice
was administrated orally with (PMFD) with 200, 400 and
600 mg/kg doses. Metformin group-6 mice was treated with The STZ-induced Balb/c mice were treated with PMFD and
metformin drug orally. Each dose of PMFD and metaformin blood glucose level was recorded at on first day before and
was diluted after 3 hours of treatment. The data on (figure 1) clearly
with 4 ml of distilled water. shows that the glucose level was decreased in all the mice of
each group. The positive control mice blood glucose level
3.2.6 Measurement of blood glucose levels was found to be increased. However, the statistical
Blood glucose level of each mice of all groups was significance test shows that only results of group 3, 4 and 5
measured by using one touch glucometer. The blood were significant and group 6 results were insignificant
samples were collected from the tail vein of mice by using (Table 3).
sterile needle . Blood glucose level of each mice was
measured before and after the treatment of diabetic mice

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Table 3: Effect of PMFD on blood glucose levels in STZ-induced Balb/c mice after 3 hours treatment (1 st day) to diabetic
mice
(An asterisk signifies a statistically significant difference change in glucose level in PMFD Balb/c mice before and after 3
hours of administration at P < 0.05.).
Figure 1: Comparison of change in blood glucose level in STZ-induced Balb/c mice before and after 3 hours of PMFD
treatment(1st day)
4.2 Result of PMFD on blood glucose levels in treatment to diabetic mice. The positive control mice blood
STZ-induced Balb/c mice 2 hours after treatment (2 nd glucose levels were found to be increased. The results show
day) to diabetic mice. that the blood glucose level in all the mice were reduced
(Figure 2). All the results found to be statistically significant
The blood glucose level was measured for each mice of all (Table 4).
groups on second day before and after 2 hours of PMFD
Table 4: Effect of PMFD on blood glucose levels in STZ-induced Balb/c mice after 2 hours treatment (2nd day) to diabetic
mice
hours of administration at P < 0.05.).

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treatment (2nd day)
4.3 Result of PMFD on blood glucose levels in levels were reduced after the treatment of PMFD in group 3,
STZ-induced Balb/c mice 2 hours after treatment (3th 4, 5 and 6 compared to before treated mice (Figure 3). The
day) to diabetic mice. positive control mice blood glucose level was found to be
increased. All the results found to be statistically significant
From the results, the effect of PMFD treatment on change in (Table 5).
glucose level in diabetic Balb/c mice before and after 2
hours of administration on third day shows that the glucose
Table 5: Effect of PMFD on blood glucose levels in STZ-induced Balb/c mice after 2 hours treatment (3th day) to diabetic mice
hours (3th day) of administration at P < 0.05.)

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treatment (3th day)
compared to before treatment mice (Figure 4). The positive
4.4 Result of PMFD on blood glucose levels in control mice blood glucose level was found to be increased.
STZ-induced Balb/c mice after 2 week’s treatment to All the group’s results show statically significant reduction in
diabetic mice blood glucose level (Table 6).
The blood glucose level in diabetic Balb/c mice after 2 weeks

of administration of PMFD shows reduction in all the groups
Table 6: Effect of PMFD on blood glucose levels in STZ-induced Balb/c mice after 2 weeks treatment to diabetic mice
(An asterisk signifies a statistically significant difference change in glucose level in PMFD Balb/c mice before and after 2 weeks
of administration at P < 0.05.).
Figure 4: Comparison of change in blood glucose level in STZ-induced Balb/c mice before and after 2 weeks of PMFD
treatment

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4.5 Result of PMFD on body weight in STZ-induced (Figure 5).Overall results prove that that PMFD treatment
Balb/c mice after 2 week’s treatment to diabetic mice. help in reducing blood glucose level and body weight in most
of the mice in all groups. Comparisons of blood glucose level
The body weight of diabetic mice was measured before and and body weight of all 18 mice compared with control group
after PMFD treatment at 2 weeks. The positive control mice (untreated mice) were done using the Student’s t-test. Most
blood glucose levels were found to be increased. (Table 7) of the results found to be statistically significant difference
shows that there was significant reduction in group 3, 4, and compared to the control (untreated seedlings) at P < 0.05).
5, but group 6 (diabetic mice were treated with metformin at
dose of 200 mg/kg) did not appear statistical significant
Table 7: Effect of PMFD on body weight in STZ-induced Balb/c mice after 2 weeks treatment to diabetic mice (An asterisk
signifies a statistically significant difference change in body weight in PMFD Balb/c mice before and after 2 weeks of
administration at P < 0.05.)
Figure 5: Comparison of change in body weight in STZ-induced Balb/c mice a before and after 2 weeks of PMFD treatment .
5. Discussion effectively reduced blood glucose levels (post prandial and

random blood glucose levels as well). From our
Diabetes is a metabolic disorder and major global health investigation, we found that diabetic mice treated with
problem worldwide. As per evaluation, one person is PMFD at the doses of 200, 400 and 600 mg/kg body weight
detected with diabetes every five second in the world significantly reduced plasma glucose levels on first day at 3
whereas one patient dies of in every 10 second (Colagiuri, hours (Table 3), second and third day at 2 hours after
2010). Most of prophetic plants contain carotenoids, treatment of diabetic mice (Table 4 and 5) and after 2 weeks
flavonoids, terpenoids, alkaloids, glycosides and can often (Table 6) . Metformin as a positive control also reduced
have anti-diabetic effects (Afrisham et al., 2015). Numerous plasma glucose levels .The levels of glucose reduction by
studies have substantiated the beneficial effects of medicinal (PMFD) were strongly comparable with metformin group or
herbs for controlling glycaemicstatus (Campbell-Tofte et al., higher than that of metformin group reduction. But in case of
2012; Abdelwahab et al., 2013; El-Abhar & Schaalar, the diabetic untreated group, the glucose levels were
2014). Scientific evaluation of Prophetic medicine increased in each time of 2, 3 days and 2 weeks after
recommended for the management of diabetes is lacking. We treatment. This indicates that in case of normal diabetes
designed a study to evaluate a formulation of Prophetic mice, the condition of diabetes was deteriorated with the
medicine for the reduction of blood glucose levels to the increasing of glucose levels without treatment. This proves
significantly lower levels. Our investigation demonstrated the effectiveness of (PMFD) as it could effectively control
that this innovative formulation of Prophetic medicine
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the diabetic disease status with effective control of blood side-effects. This result proves that diabetic mice treated with
glucose levels in the treated group. PMFD was significantly able to reduce fasting plasma
glucose FPG and post-prandial blood glucose PPBG levels.
Many similar previous studies have shown that cinnamon, The outcomes from this study could be contribute for the
black seed, olive leaves and ginger had effects for treatment management of plasma blood glucose of STZ- induced
of diabetes. The therapeutic potentials of α-lipoic acid diabetic Balb/c mice or minimizing the side-effects of
(α-LA), L-carnitine and N. sativa or combination of them in diabetic medication. Thus, it can be used as a raw data for
carbohydrate and lipid metabolism was evaluated in a rat manufacturing new drug for the effective management of
model of diabetes which was induced by single i.p. injection diabetes improving the quality of life for diabetic patients.
of streptozocin (STZ) 65 mg/kg. For evaluation of glucose Development of new medications from natural sources
metabolism, fasting blood glucose, insulin, insulin especially from Prophetic medicine sources will be very
sensitivity, HOMA, C-peptide, and pyruvate dehydrogenase useful as can ensure the long-term affectivity of antidiabetic
activity were determined. Either α-LA or N. drugs, minimizing or diminishing apparent side-effects.
sativa significantly reduced the elevated blood glucose level.
The combination of 3 compounds significantly increased the 7. Recommendations and Further Studies
level of insulin and C-peptide. Combination of α-LA,
L-carnitine and N. sativa will contribute significantly in Based on the findings of this study the following
improvement of the carbohydrate metabolism in diabetic recommendations are suggested:
rats, thus increasing the rate of success in management of
DM (Salama, 2011). The stretpozotocin induced diabetic in The formulation of Prophetic medicine (PMFD) can be used
rat model has demonstrated the ability of cinnamon to for the management of diabetes. However, further in vivo
normalize glucose metabolism, lipid abnormalities and studies to assess the active of the formulation on insulin,
weight changes associated with diabetes. Most of the trials on lipids, HbA1c levels may confirm its therapeutic
animal studies showed the ability of cinnamon to reduce effectiveness. Besides, clinical studies of this formulation on
FPG, postprandial glucose and or HbA1c (Subash Babu et diabetic patients for long period are also recommended. The
al., 2007; Anand et al., 2010; Shihabudeen et al., present study on formulation of Prophetic medicine can be
2011). Previous study used cinnamaldehyde extracted recommended for the prevention and treatment of diabetes.
from Cinnamomum zeylanicum to demonstrate a significant
reduction of plasma glucose and HbA1c levels in
References
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Author Profile
Eman Elaswed, received the ( higher professional diploma)
degrees in Branch of Medical Analysis from The higher Institute
for medical Professions at AlJMAIL in the academic year
2009/2010. She pursued study for the Master’s degree in
Biotechnology in the Department of Biotechnology in Lincoln
University College, Malaysia, and this paper is part of her thesis
results in Biotechnology.

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International Journal of Science and Research (IJSR)

ISSN (Online): 2319-7064

Evaluation of Antidiabetic Potential of

1. Introduction The aim of the present study is to investigate or evaluate the

3.1.2.Chemicals cinnamon powder to 500 ml of 80 % ethanol and by shaking

Volume 6 Issue 3, March 2017

Volume 6 Issue 3, March 2017

Volume 6 Issue 3, March 2017

The blood glucose level in diabetic Balb/c mice after 2 weeks

Volume 6 Issue 3, March 2017

5. Discussion effectively reduced blood glucose levels (post prandial and

Volume 6 Issue 3, March 2017

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