Antidiabetic and Hepatoprotective Activities of
Bombax ceiba Young Roots in Alloxan-Induced
Rokshana Sharmin1, Maruf-ul-Islam1, Md. Hasibul Hasan Joarder1, Md. Mohiuddin Alamgir1,
Md. Golam Mostofa2 and A. H. M. Khurshid Alam2*
1Department of Pharmacy, Jessore University of Science and Technology, Jessore-7408, Bangladesh
2Department of Pharmacy, University of Rajshahi-6205, Bangladesh
A. H. M. Khurshid Alam, Department of Pharmacy, University of Rajshahi, Rajshahi-6205, Bangladesh, Tel: +880721711110 (office); Fax: +
Received: June 14, 2018; Accepted: August 2, 2018; Published: August 31, 2018
Khurshid Alam AHM, Sharmin R, Maruf I, et al. (2018)Antidiabetic and Hepatoprotective Activities of Bombax ceiba
Young Roots in Alloxan-Induced Diabetic Mice. J Nutrition Health Food Sci 6(5):1-7. DOI: 10.15226/jnhfs.2018.001140
Bombax ceiba (B. ceiba), a member of the Malvaceae family, has
widely been used by rural practitioners for various ailments from
ancient civilization. Although different parts of this plant are known
to have several biological activities, very little is known about the
effects of young roots of B. ceiba (BCYR) on diabetes and hepatic
toxicity. Therefore, the present study was undertaken to investigate
the effects of the BCYR on diabetes and hepatic toxicity in Alloxan-
Induced Diabetic Mice (AIDM). The dried coarse powder of the BCYR
was extracted with ethanol (Et-BCYR) by cold extraction method.
Qualitative phytochemical screening of the Et-BCYR revealed
the presence of flavonoids, phenolics, tannin, steroids, alkaloids
and glycosides. Administration of the Et-BCYR (400 mg/kg bw)
intraperitoneally in AIDM, it significantly (p>0.05) reduced the blood
glucose level compared to untreated AIDM at different time points
(0-24 hours). Strikingly, the most action was 78.36% in 16 hours,
which was higher than that of the standard met for min (72.36%).
Treatment with Et-BCYR significantly (p>0.0001) elevated the HDL
level; on the contrary, it reduces LDL, TC and TG levels when compared
to untreated AIDM. Additionally, Et-BCYR treatment significantly
(p>0.0001) decreased the hepatotoxicity by detecting the reduced
level of SGOT and SGPT (hepatotoxic markers) compared to untreated
AIDM. Our findings show that the young roots of B. ceiba have potential
hypoglycemic, hypolipidemic and hepato protective activities and
confirm the traditional uses of this plant to manage diabetes and its
associated liver toxicity.
Keywords: Diabetes mellitus; Bombax ceiba; Hypoglycemic;
• BCYR= Bombax ceiba Young Root
• Et-BCYR= Ethanolic extract of Bombax ceiba Young Root
• SGPT = Serum Glutamate Pyruvate Transaminase
• SGOT = Serum Glutamic Oxalate Transaminase
• FBG=Fisting Blood Glucose
• TC= Total Cholesterol
• TG= Tri Glyceride
• LDL= Low Density Lipoprotein
• HDL= High Density Lipoprotein
Diabetes is a chronic disorder of carbohydrate, fat and protein
metabolism, which is characterized by high blood sugar level due
to either inadequate synthesis of insulin or insulin resistance
. It is the most leading causes of death globally and more
than 25% of the total population are affected with this disease,
and it will be increased to 50% by 2025 . Nisha et al., (2014)
reported that hyperlipidemia is one of the major complications
of diabetes mellitus due to abnormal lipid metabolism . In
chronic hyperglycemia, excess formation of free radicals leads
to the development of the diabetic nephropathy . The higher
level of lipid profile, especially Total Cholesterol (TC) synthesis
during hyperglycemic condition contributes to accelerate the
atherosclerosis in diabetes mellitus . The International
Diabetes Federation (IDF) estimated more than 7.1 million
people suffer from diabetes in Bangladesh and an equal number
of people remain unknown with their diabetic condition and the
number of affected people will be 15 million by 2025. For this,
treatments, created along the standards of western prescription
(allopathic) are regularly constrained inadequacy, carry the
risk of adverse effects and are often too costly, particularly in
the developing world. Thus, it is essential to discover a potent,
safe and cost-effective hypoglycemic agent. Since ancient time,
diabetes treatment has been done orally with several medicinal
plants or their extracts based on their folkloric reputation.
Recently, WHO has also been recommended to use of traditional
plants as a treatment for diabetes patient . For the last few
years, herbal medicine and herbal drugs used were gradually
increasing in developing and developed countries . Hence, the
search for potent pharmacologically active agents from natural
sources such as from medicinal plants or their extracts that have
led to the discovery of many clinically useful drugs might play a
major role in the treatment of human diseases . Keeping this
in mind, we have chosen a plant, Bombax ceiba (B. ceiba), which
has different biological activities, including hypoglycemic activity.
B. ceiba, a member of the Malvaceae family, is a lofty, deciduous
tree. It is popularly known as silk cotton tree and widely distributed
in temperate and tropical Asia, Africa and Australia. Different
parts of this plant are reported to have therapeutic potentials
against different diseases such as diabetes, hepatic toxicity,
infections, asthenia, polyurea and glycosuria [9-12]. Although
different parts of B. ceiba are known to have different biological
activities including antioxidant, antimicrobial, anti-inflammatory
and analgesic, ,hypotensive and hypoglycemic activity, very little
work has been done by taking an ethanolic extract of young roots
of B. ceiba (BCYR) on diabetes and hepatic toxicity [9,10,13-15].
Therefore, the present study was undertaken to investigate the
effects of the BCYR on diabetes and hepatic toxicity in Alloxan-
Induced Diabetic Mice (AIDM).
Materials and Methods
According to Ayurveda, B. ceiba has proven medicinal
properties and is the ingredient of many formulations. The roots
are sweet, cooling, stimulant, restorative, astringent, alternative,
aphrodisiac, demulcent, emetic and tonic and they are used in the
treatment of diarrhea, hepatotoxicity, dysentery, menorrhagia,
styptic and for wounds [16-20]. The chemical investigations
worldwide suggest that the roots of B. ceiba are used for the
medicinal purpose because they are rich in lupeol, β-sitosterol
and sesquiterpenes, which are beneficial for health in treating
some diseases. Apart from the above active constituents, the stem,
root, flower, fruit and leaves of B. ceiba have been reported to
contain many important phytoconstituents, including alkaloids,
glycosides, flavonoids, steroids, saponins, phytosterols and
triterpenoids (lupeol and beta-sitosterol), phenolic compounds
and tannins . From ancient civilization, tips of young root
have been used as a vegetable for patients suffering from
impotency. Jain et al., (2009) reported that young roots of B. ceiba
(BCYR) are used traditionally as an ethnomedicine for various
ailments, including calculous affections, chronic inflammation
and ulceration of the bladder and kidneys. Moreover, younger
roots of B. ceiba are more nutritious than the older ones and are
roasted in the fire to eat like roasted sweet potato during famine
or otherwise also . Based on above mentioned reasons, young
roots instead of adult roots were selected for this study. The
fresh BCYR were collected from rural area of Kustia and Jessore
district in Bangladesh. The species was identified by an expert
taxonomist of Bangladesh national herbarium, Dhaka. After
collection, the fresh roots were thoroughly washed with distilled
water and kept apart barking layer from the roots and sliced into
small pieces. The roots were sun dried under shade and ground
with an electric grinder into coarse powder and were used for
Preparation of ethanolic extract
The dried coarse powders were soaked with 1.5 L of 95%
ethanol in amber-coloured extraction bottle. The bottles
were sealed and kept for the 7 days at room temperature with
occasional shaking and stirring. The extracts were filtered
through cotton followed by Whatman No.1 filter paper and
were concentrated with a rotary evaporator (Bibby Sterlin Ltd,
UK) under reduced pressure at 50°C to afford brown-coloured,
powdery crude extract.
Qualitative Phytochemical Profiles
Test for Carbohydrate (Benedict’s test)
To 5 ml of Benedict’s reagent, 1 ml of Et-BCYR solution was
added and boiled for 2 minutes and cooled. Formation of a red
precipitate shows the presence of carbohydrate.
Test for Saponins (Foam test)
Ten (10) mg of Et-BCYR was taken in a test tube and shaken
vigorously with 5 ml of water. Production of persistent foam
indicates the presence of saponins.
Test for Flavonoid, Phenol and Tannin (Ferric chloride
Five (5) mg of Et-BCYR was dissolved in 2 ml of water. 1 ml of
neutral 5% ferric chloride solution was added. A dark green color
indicates the presence of flavonoid, phenols and tannin.
Test for Proteins (Biuret Test)
Three to five (3-5) mg of Et-BCYR was added in 4% NaOH and
few drops of 1% CuSO4 solution was added in this solution. A
violet or pink color indicates the presence of protein.
Test for Steroids (Liebermann-Burchard test)
The Et-BCYR was dissolved in 1 ml of chloroform. 2 ml acetic
anhydride and 1 ml concentrated sulfuric acid were added.
Formation of greenish color solution indicates the presence of
Test for Phytosterol (Salkowski’s reaction)
To 2 ml of Et-BCYR, 2 ml of chloroform and 2 ml of concentrated
H2SO4 were added and shaken vigorously. Chloroform layer shows
greenish yellow fluorescence, which indicates the presence of
Test for Amino Acids (Ninhydrin test)
To an aliquot of diluted extract, 2 ml of ninhydrin solution was
added. A violet color formation indicates the presence of amino
Test for Glycosides (Keller- Killani Test)
About 2 ml of extracts was taken in a test tube followed by 1
ml of glacial acetic acid containing trace amount of FeCl3 and 1
ml of concentrated H2SO4 were added to the extract carefully. A
reddish-brown colour is formed at the junction of two layer and
the upper layer turns bluish green in presence of glycosides.
Test for Alkaloids (Wagner’s Test)
The Et-BCYR was taken in a test tube. 5 ml of 1% HCl was
added to the test tube and stirred on a stream bath. After filtering
the solution, few drops of Wagner’s reagent was added. Formation
of a reddish brown precipitate indicates the presence of alkaloids.
Preparation of Dose
The dose (120 mg/kg body weight) of standard metformin
is prepared by using sterilized water. BCYR extract is dissolved
in 10 % DMSO to prepare dose at a concentration of 400 mg/kg
body weight, bw.
Induction of Diabetes
For the development of diabetic model mice, the mice were
grouped into 4 classes (Group I-IV). All the mice in group II-IV
were kept overnight fasting and a freshly prepared solution of
alloxan monohydrate (150 mg/kg bw in 0.9% normal saline) was
administered intraperitoneally. The group I mice was kept as a
normal control group. After 48 hours of alloxan induction, blood
glucose content was measured by a Glucometer (SAFE TOUCH
Glucometer, HMD Bio Medical Inc., Taiwan Technology of USA).
Mice with blood glucose levels above 11.1 mmol/L were selected
for the study. Baseline blood glucose level of group II-IV mice was
also measured just prior to the administration of alloxan.
Grouping of Experimental Mice
Swiss Webster mice were randomly assigned into five groups
Group 1 (Normal control) received 0.9% normal saline (1 mL/
Group 2 (Diabetic control) mice received alloxan in 0.9% normal
Group 3 (positive control, standard) mice received metformin
hydrochloride (120 mg/kg, bw) in 0.9% normal saline.
Group 4 (Et-BCYR 400) mice received Et-BCYR extract (400 mg/
kg, bw) in 10% DMSO.
The protocol used in this study for the use of rat as an animal
model for diabetes research was approved by the Rajshahi
University Animal Ethical committee (27/08/RUBCMB). This
research work was approved by the Ethical Review Committee of
Research Cell of Rajshahi Medical College, Bangladesh (ref. RMC/
Blood samples were withdrawn by cutting the tail-tip of each
group of mice on the 0, 4, 8, 12, 16, 20 and 24 hours of the day
for estimating the blood glucose level. At the end of treatment,
the mice were sacrificed and the levels of HDL, LDL, TG, TC, SGOT
and SGPT were determined using commercial kits according to
the manufacturer protocols by a semi-auto analyzer.
Data were expressed as mean ± standard error of mean (SEM).
Statistical comparisons were performed by two-way analysis of
variance (ANOVA) for blood glucose analysis and two–way ANOVA
analysis. The results were considered to be significant when p
values were less than 0.05 (p< 0.05). Statistical calculations and
graphs were prepared using Graph Pad Prism Version 6.00 for
Windows (Graph Pad Software, San Diego, CA, USA).
Phytochemical Screening of Et-BCYR Extract
Freshly prepared ethanolic extract of the BCYR (Et-BCYR)
was subjected to preliminary phytochemical screening for
various constituents. Table 1 shows the differential distribution
of phytoconstituents in the Et-BCYR extract. Et-BCYR contains
large amounts of flavonoids, phenolics, tannins, and steroids. It
also contains cardiac glycosides and alkaloids in the moderate
Effect of Et-BCYR on Blood Glucose Level in Alloxan-
Induced Diabetic Mice (AIDM)
The Et-BCYR (400 mg/kg bw), and the standard metformin
(150 mg/kg bw) were administered i.p. in AIDM and the blood
samples were collected on the 4, 8, 12, 16, 20 and 24 hours of
treatment. Et-BCYR extract significantly reduced the blood
glucose levels at different time points (4, 8, 12, 16, 20 and 24
hours) when compared with the untreated AIDM. Interestingly,
the reduction activity of glucose level in blood was the maximum
of 16 hours and found to be 78.36%, whereas the standard
metformin reduced the blood glucose level to 72.36% by 16 hours
suggest that Et-BCYR has higher hypoglycemic activity than that
of a standard (Figure 1). All the experiments were done in three
different times and values are expressed as mean ± SEM (n=3).
Figure 1: Effect of Et-BCYR on blood glucose level in AIDM. * indicates
significant change (p< 0.0001) of blood glucose level in Et-BCYR-treated
AIDM compared to untreated AIDM. All the experiments are carried out
in three individual time and the results are expressed as means ± SEM
Effect of Et-BCYR on Lipid Profile in AIDM
After the measure of glucose level in blood, the mice were
sacrificed and blood samples were collected to measure the lipid
profile by a semi bio-analyzer. The standard metformin reduced
the Total Cholesterol (TC) level to a maximum of 76.69% when
compared to AIDM. On the other hand, treatment with Et-BCYR,
in comparison to AIDM, reduced the TC level to 78%, which was
higher than that of standard metformin (76.69%) (Figure 2A).
The treatment of the Et-BCYR and the standard metformin
reduced the Triglyceride (TG) level to 75.24% and 79.81%,
respectively in AIDM when compared with the untreated AIDM.
The activity of the Et- BCYR was higher than that of the standard
drug (Figure 2B). Beside these, Et-BCYR reduced the low lipid
density (LDL) level to 92.71%, on the other hand, it significantly
increased the high density lipoprotein (HDL) to 255.78%, which
was higher than standard metformin (204.95%) (Figure 2C-2D).
All the results together suggest that the Et-BCYR shows a potent
dislipidemic activity. All the experiments were done in triplicate
and values were expressed as mean ± SEM (n=3).
Figure 2: Effect of Et-BCYR on lipid profile in AIDM. * indicates significant change (p< 0.0001) of TC (Fig. 2A), TG (2B), LDL (Fig. 2C) and HDL (Fig. 2D) levels in Et-BCYR-treated AIDM compared to untreated AIDM. All the experiments are carried out in three individual times and the results are expressed as means ± SEM (n=3).
Effect of Et-BCYR on Hepatic Toxicity in AIDM
The level of hepatotoxicity markers, Serum Glutamic Pyruvate
Transaminase (SGPT) and Serum Glutamic Oxalate Transaminase
(SGOT) in AIDM and treated AIDM were examined by a semi bioanalyzer.
Et-BCYR extract and the standard metformin declined
the SGOT and SGPT level to 58% and 81.11%, respectively (Figure
3A-3B). Values are expressed as mean ± SEM of 3 independent
Figure 3: Effect of Et-BCYR on liver toxicity in AIDM. * indicates significant change (p< 0.0001) of SGOT (Fig. 3A) and SGPT (Fig. 3B) levels in Et-BCYR treated AIDM compared to untreated AIDM. All the experiments are carried out in three individual times and the results are expressed as means ± SEM (n=3).
Alloxan monohydrate destroys the β-cells of islets of
Langerhans of the pancreas and inhibits the production of
insulin, which affect to push glucose into the body tissues,
resulting high level of glucose, decreased protein content and
increased cholesterol and triglyceride level in the blood . In
this study, alloxan (150 mg/kg bw in 0.9% normal saline) was
administered intraperitoneally in mice and the mice, which have
a base line blood glucose level above 11 mmol/L was selected for
the study. The treatment with standard metformin reduced blood
glucose level of 5.58 mmol/L, whereas the Et-BCYR significantly
reduced blood glucose level to 5.38 mmol/L, (Figure 1), which
was similar to that of the standard suggesting that the extract
might control diabetes by increasing the production of insulin.
Qualitative phytochemical analysis of the Et-BCYR revealed the
presence of alkaloid, glycoside, tannins, and flavonoids (Table1)
(Rani N, 2012) reported that flavonoids put down glucose level
significantly by inhibiting α-glucosidase enzyme . Generally
alkaloids inhibit α-glucosidase enzyme and decrease glucose
transport through the intestinal epithelium cell . In this
study, we assume that phytochemicals, which have been detected
in Et-BCYR, reduced glucose level by inhibiting the α-glucosidase
enzyme leading to decrease glucose transport through the
Table 1: Phytochemical constituents of ethanolic extract of young roots of B. ceiba (Et-BCYR)
No. of Tests
Name of Tests
Name of Phytochemicals
Ferric chloride test
Ferric chloride test
Phenolic compound & Tannin
Fixed oils & fatty acid
- = absent; + = present in mild amount, ++ = moderate amount; +++ = large amount
Our results are consistent with those of previously
reported data . The most common lipid abnormalities in
diabetes are hypertriglyceridemia and hypercholesterolemia
. Hypertriglyceridemia is also associated with metabolic
consequences of hypercoagulability, hyperinsulinemia, insulin
resistance and glucose intolerance . Administration of the
Et-BCYR significantly (p< 0.0001) reduced the TG (Figure 2A) and
TC (Figure 2B) levels suggest that the observed hypolipidemic
effect was due to decreased cholesterologenesis and fatty acid
synthesis . Moreover, the Et-BCYR treatment significantly (p
< 0.0001) reduced LDL level (Figure 2C) and increased HDL level
(Figure 2D). These values are a very desirable in the biochemical
state for prevention of atherosclerosis and ischemic conditions
. Accumulating evidence suggests that a decrease in the
ratio of the TC/HDL cholesterol (atherogenic index) lessens the
risk of heart disease. Previous studies demonstrated that
phytochemicals have the capacity for lipid-lowering properties
[30&31]. Flavonoids prevent the oxidation of LDL, lower the TC
and TG levels leading to minimize the risk of atherosclerosis
[32&33]. Cardiac glycosides also used as diuretics and heart
tonics because of its valuable effects on the heart and affect
the availability of intracellular Ca2+for myocardial contraction
. SGPT is a better parameter than SGOT to identify the liver
toxicity, since SGOT also found in kidney and cardiac muscle
. The Et-BCYR contains high level of flavonoids and phenolic
compounds, which are natural antioxidants. They can scavenge
off free radicals. So, the anti-oxidant principles may be involved
in the hepatoprotective activity. In this study, Et-BCYR reduced
SGOT (Figure 3A) and SGPT (Figure 3B) to 58% and 76.53%,
respectively suggest that Et-BCYR can be considered as an agent
for the treatment of liver toxicity.
The present study clearly indicates that the young roots of
Bombax ceiba possess prolong and potential hypoglycemic activity
evidenced by 24 hours. The effect of this plant on biochemical
alterations reveals hypolipidemic and hepatoprotective
activities and confirms the traditional uses of this plant in the
management of diabetes and its associated liver toxicity. The
phytochemicals present in this plant might be account for the
observed pharmacological actions. Further study is necessary
to characterize the potential compound(s) and their role in
the control and management of diabetes and hepatotoxicity in
Conflict of Interests
The author(s) declare(s) that there is no conflict of interests
regarding the publication of this article.
The authors want to thank Department of Pharmacy, Jessore
University of Science and Technology and Ethno-pharmacology
Lab, Jahangir Nagar University, for providing necessary facilities
to carry out this research work.
The authors declare that they have no fund for this research.
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