European Journal of Medicinal Plants

8(2): 78-86, 2015, Article no.EJMP.2015.092

ISSN: 2231-0894

SCIENCEDOMAIN international
www.sciencedomain.org

Hypoglycemic and Hypolipidemic Potentials of

Extract of Vernonia calvoana on Alloxan-Induced
Diabetic Albino Wistar Rats
I. A. Iwara1*, G. O. Igile1, F. E. Uboh1, E. U. Eyong1 and P. E. Ebong1
1

Department of Biochemistry, Faculty of Basic Medical Sciences, University of Calabar,

Calabar, Nigeria.
Authors contributions
The sole author designed, analyzed and interpreted and prepared the manuscript.
Article Information

DOI: 10.9734/EJMP/2015/16058
Editor(s):
(1) Marcello Iriti, Department of Agricultural and Environmental Sciences, Milan State University, Italy.
Reviewers:
(1) Abdullahim Nuhu, Kaduna Polytechnic, Kaduna, Nigeria.
(2) Musa Yakubu Tula, Federal Polytechnic Mubi, Adamawa State, Nigeria.
(3) Anonymous, Romania.
Complete Peer review History: http://www.sciencedomain.org/review-history.php?iid=1084&id=13&aid=8870

Original Research Article

Received 3rd January 2015

th
Accepted 16 January 2015
Published 17th April 2015

ABSTRACT
Objective: Investigations were carried out to evaluate the therapeutic potentials of extract of
Vernonia calvoana as herbal medicines.
Methods: In this study, the hypoglycemic and hypolipidemic potentials of extract of Vernonia
calvoana was evaluated by using normal and alloxan-induced diabetic rats at a dose of 400 mg/kg
b.w. daily for 14 days. Fasting Blood glucose levels and body weight were monitored at 7 days
intervals, and different biochemical parameters, serum glucose, serum cholesterol, serum
triacylglycerol, high density lipoprotein, low density lipoprotein and very low density lipoprotein were
also assessed in the experimental animals.
Results: It was found that extract of Vernonia calvoana showed significant hypoglycemic and
hypolipidemic activity (P<0.01).
Conclusion: Extracts of Vernonia calvoana showed significant hypoglycemic and hypolipidemic
activity, which provides the scientific proof for their traditional claims.

Keywords: Vernonia calvoana; hypoglycemic; hypolipidemic and alloxan.

_____________________________________________________________________________________________________
*Corresponding author: E-mail: iwaraiwara83@gmail.com;

Iwara et al.; EJMP, 8(2): 78-86, 2015; Article no.EJMP.2015.092

medicinal and industrial uses. For example,

Vernonia amygdalina, Vernonia calvoana and
Vernonia colorata, are eaten as leafy vegetables
[9,10]. Vernonia species frequently used in
ethnomedicine include Vernonia amygdalina,
Vernonia condensata, Vernonia cineria, Vernonia
guineensis and Vernonia conferta. Vernonia
galamensis are used industrially for its seed oil
contents. Vernonia amygdalina is the most
studied member of the Vernonia genus as well
as one of the most studied plants in Africa [11].
Despite the widespread use of plants of the
Vernonia genus in food and medicine and a
comprehensive review on the nutritional and
health of Vernonia genus by Toyang and
Verpoorte [12] in human and animal subjects,
Vernonia calvoana Hook. F. (V.C) is amongst the
less known species of the genus. The ethnomedicinal uses of this species as reported by
Focho et al. [13] include the relief of navel aches
and constipation. However, recent reports have
shown that this plant contains high bioactive
component with high nutrient composition which
contributes to the nutrient intake by the
consuming population [14]. To ll this gap, this
research is designed to provide knowledge on
the hypoglycemic and hypolipidemic evaluation
of this Vernonia specie, conrm claims of their
traditional uses and further research into its
development of medicines.

1. INTRODUCTION
Chemotherapy and chemoprophylaxis which
form an important and major aspect of disease
control in Africa and the world over, is beset with
problems which include resistance to drugs,
drug toxicity, protracted treatment protocol and
limited repertoire of compounds [1]. The problem
of drug toxicity and resistance is very
pronounced in the management of diabetes.
However, in a medicinal plant, several agents
(phytochemicals) may exert a wholistic action in
various mechanistic targets, hence a better
option.
Dependence on medicinal plants as medicines in
the treatment of disease is common among a
large proportion of population of the rural
populace because of its availability and
affordability [2]. Due to the increase in
awareness of the importance of traditional
medicine in human and animal healthcare,
researches into the efficacy of some of the herbs
used in the treatment of some illness would be
worthwhile. The report of WHO, [3] supports the
use of effective and safe remedies, and accepts
traditional medicine as a valuable resource, for
primary healthcare. Medicinal plants constitute a
fundamental component of primary traditional
healthcare system in rural communities
throughout Africa and beyond. According to
Odugbemi, 2008 [4] in the last twenty years,
medicinal plants have received huge attention
both in developed and developing countries [5].
However, medicinal plants that are commonly
used for the treatment of many diseases, include,
Eucalyptus globules (blue gume), Momodica
charantia, Azadirachta indica, Gongronema
latifolium, Vernonia amygdalina etc and extracts
obtained from these leafy vegetables are used to
manage some ailments or diseases [6].

2. MATERIALS AND METHODS

2.1 Plant Materials
2.1.1 Collection of plant materials
Vernonia
calvaona
(Ekeke)
green-leafy
vegetable was purchased from Ugep in Yakurr
LGA of Cross River State of Nigeria. The plant
sample was authenticated by Dr. Michael Eko, a
Botanist in the University of Calabar, Nigeria, and
a voucher specimen (BCH/VC/01) was deposited
in the Herbarium of the Department of
Biochemistry, University of Calabar, Nigeria.

It is observed in most developing countries that

attention has been focused on use of medicinal
plants and studies on medicinal plants however
have concentrated on plant extracts [7].

2.2 Sample Preparation

Vernonia (Asteraceae) is the largest genus in the

tribe Vernoniae, with close to 1000 species [8].
Vernonia species grow in a wide range of
habitats of broad ecological diversity and climatic
conditions including tropical forest, marshes and
wet areas, dry plains; tropical savannahs dry
sites and even frosty regions of eastern part of
North America [8]. The genus is morphologically
made up of annuals, herbaceous perennials
lianas, shrubs, and trees. The genus Vernonia is
known for having several species with food,

The leaves were washed, cut into small pieces

and air-dried at room temperature (272C) for
seven days. The dry leaves were blended with
the use of cornono (El legitima) V C.I.A. S.A
manual hand blender, Made in Colombia
(Medellin-Colombia) into powdered and 1.5 kg of
the blended leaves was weighed out and soaked
in 2000 ml of a solution of Methanol and
Dichloromethane (1:1, v/v). The mixture was

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Iwara et al.; EJMP, 8(2): 78-86, 2015; Article no.EJMP.2015.092

allowed for 48hours in the refrigerator at 4C for

thorough extraction of the plants active
components. This was then filtered with a chess
cloth material and later with Whatman No. 1 filter
paper to obtain a homogenous filtrate. This
filtrates were then concentrated in vacuo at low
temperature (37-40C) to about one tenth the
original volume using a rotary evaporator. The
concentrates were allowed open in a water bath
(40C) for complete dryness. The extract was
then refrigerated at 2- 8C until when used.

dose of 400 mg/kg body weight. The control

animals received 0.2 ml of normal saline
(placebo). The treatment lasted for 14 days and
throughout this periodic changes in blood
glucose and body weight were measured with
the use of a glucometer and electronic weighing
balance respectively. The animals were
maintained on pellets prepared with Growers
feed from Vital Feeds, Jos, Plateau state,
Nigeria, and tap water. Both the feed and water
were provided ad libitum.

2.3 Animals

2.7 Collection of Samples for Analysis

Twenty four (24) albino rats (both sexes) of

wistar strain weighing about 140-180 g were
obtained from the animal house of the
Department of Biochemistry, University of
Calabar, Calabar. The animals were allowed to
acclimatize for two weeks in the animal house of
the Department of Biochemistry. The animals
were housed in well ventilated cages (wooden
bottom and wire mesh top) and kept under
controlled
environmental
conditions
of
temperature (255C), relative humidity (505%)
and 12 hour light/dark cycle. The protocol was in
accordance with the guidelines of the National
Institute of Health (NIH) publication (1985) for
laboratory Animal Care and Use and approved
by the Collage of Medical Sciences Animal
Ethics Committee University of Calabar, Nigeria.

At the end of the 14 days, food was withdrawn

from the rats and they were fasted overnight but
had free access to water. They were then
euthanized under chloroform vapor and
sacrificed. Whole blood was collected via cardiac
puncture using sterile syringes and needles. The
blood was emptied into plain tubes and allowed
to clot for about two hours. The clotted blood was
thereafter centrifuged at 3,000 rpm for 10
minutes to recover serum from clotted cells.
Serum was separated with sterile syringes and
needles and stored frozen until used for
biochemical analysis.

2.8 Data and Statistical Analysis

The results were analyzed for statistical
significance by student t-test using SPSS
software. All data were expressed as mean
SEM.

2.4 Experimental Design

The design consisted of twenty-four (24) rats
divided into four (4) groups consisting of normal
and diabetic treated group. The dose used was
based on the predetermined LD50 values
obtained from preliminary studies using Lorkes
method 1983 [15].

2.9 Animal Grouping

Schedule
Group

2.5 Induction of Experimental Diabetes

Hyperglycemia was induced by a single
intraperitoneal injection of 100 mg/kg of alloxan
(SigmaAldrich, Inc, St. Louis, MO 63103,
U.S.A.) in sterile saline. After 3 days of alloxan
injection, the hyperglycemic rats (glucose level >
180 mg/dl) were separated and divided into
different groups comprising of 6 rats each for the
anti-diabetic study.

and

Normal control
Diabetic control
Diabetic treated

Number of
animals
6
6
6

Diabetic treated

Treatment
Treatment
Placebo
Placebo
Insulin
(5IU/kg b.w)
V. calvaona
extract (400
mg/kg b.w)

2.10 Measurement of Fasting Blood

Glucose and Body Weight Change
Fasting blood glucose was determined at 7 days
interval during the 2 week experimental period
using glucucometer (one touch) and body weight
determined using electronic weighing balance.

2.6 Extract and Drug Administration

Before administration, the extracts were
reconstituted in normal saline (vehicle) and
administered orally via gastric intubation at a

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Iwara et al.; EJMP, 8(2): 78-86, 2015;; Article no.EJMP.2015.092

no.

V.C, the diabetic rats showed

owed a significant
(P<0.05) increase and body weight gain
compared to the normal control.

2.11 Blood Sampling and Biochemical

Analysis

Estimation of serum blood glucose level:

level
Serum blood glucose level was estimated
enzymatically using AGAPPE
PE diagnostic kit
(Switzerland) according to manufacturers
instruction.
Estimation of serum lipid profile level:
Serum lipid profile viz triacylglycerol (TG),
total cholesterol (TC), low density lipoprotein
cholesterol (LDL), High density lipoprotein
cholesterol
(HDL-C)
C)
was
estimated
colorimetrically using assay kit method from
AGAPPE diagnostic (Switzerland) according
to manufacturers instruction.

Figs. 4, 5, 6, 7 and 8 show the effect of 14 days

administration of extract of Vernonia calvoana on
stz induced diabetic albino wistar rat. Observed
was a significant
nt (p<0.05) increase in lipid profile
viz: Total cholesterol (TC), Triacylgycerol (TG),
High density lipoprotein-cholesterol
cholesterol (HDL-C),
(HDL
Very low density lipoprotein cholesterol (VLDL-C)
(VLDL
and Low density lipoprotein cholesterol (LDL
(LDL-C)
in the alloxan induced diabetic
iabetic rats compared to
the normal. On treatment with extract for 14
days, significant (p<0.05) decrease in TC, TG,
HDL-C, VLDL-C and LDL-C
C levels in all treated
animals was observed compared to both diabetic
and normal control rats. However, significant
decrease was also observed in TC and LDL
LDL-C of
the Vernonia calvoana treated group compared
to the insulin treated groups.

3. RESULTS
Figs. 1 and 2 show the effect of 14 days
administration of extract of Vernonia. calvoana
(V.C) on the fasting blood and serum glucose
level of alloxan- induced diabetic rats. It was
observed in the study that, the concentration of
fasting blood glucose was significantly (P<0.05)
increased in the alloxan-induced
induced diabetic rats in
the study. On administration of the extract of V.C,
the fasting and serum glucose concentration
significantly (p<0.05) reduced compared to the
diabetic control group. The result also showed
that the pattern of reduction progresses as the
experimental day.

4. DISCUSSION
Increase in fasting and serum blood glucose was
observed in alloxan induced diabetic rats which
wh
was in line with Atangwho et al., 2007 [16],
Ebong et al., 2008 [17], who also has reported
increase glucose concentration at diabetic
condition. Alloxan is a diabetogenic agent with
two distinct pathological effects interfering with
the physiological function of the pancreatic beta
cells and causes a state of insulin dependent
diabetes mellitus through its ability to inhibit the
glucokinase, the glucose sensor of the beta cell
and cause a state of insulin dependent
diabetes mellitus through its ability
lity to induce a
selective necrosis of the beta cells [18].

Fig. 3 revealed
ed the effect of extract of V.C on the
body weight change of alloxan induced diabetic
rats. It was observed in the study that alloxanalloxan
induced diabetic rats had a reduction in body
weight. However, on administration of extract of

Fig. 1. Comparison of serum glucose concentrations in the different experimental groups

Values
ues are expressed as mean + SEM; n = 4; a = p<0.05 vs diabetics; b = p<0.05 vs insulin

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Fasting blood glucose conc. (mg/dL)

Iwara et al.; EJMP, 8(2): 78-86, 2015;; Article no.EJMP.2015.092

no.

NC
Diabetics
Insulin
VC
500
450
400
350
300
250
200
150
100
50
0

*, b

*
*, b

Basal

Day 7

Day 14

Variable
Fig. 2. Comparison of fasting blood glucose concentrations
of the different experimental groups.
Values are expressed as mean + SEM, n = 4.
*significantly different from NC at p<0.05;
a = p<0.05 vs diabetics;
b = p<0.05 vs insulin.

Fig. 3. Comparison of body weight of the different experimental groups

Values
ues are expressed as mean + SEM; n = 4; *significantly different from NC at p<0.05;
a = p<0.05 vs diabetics; b = p<0.05 vs insulin

On administration
serum glucose
(p<0.05) reduced
control group. The

of the extract of V.C,

V.C the
concentration significantly
compared to the diabetic
findings are consistent with

earlier studies done by Atangwho et al. [16] and

Ebong et al. [17] on Vernonia amygdalina when
administered to diabetic animals. R
Report by Igile
et al. [14] revealed the presence sterols,

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Iwara et al.; EJMP, 8(2): 78-86, 2015;; Article no.EJMP.2015.092

no.

saponins,
coumarins,
quinones,
tannins,
flavanoids. These principles are known to be
bioactive for the management of diabetes. It is
well known that certain flavonoids exhibit
hypoglycemic activity and pancreas beta cell
regeneration ability. Sterols have also shown to
decrease blood sugar in experimental animal
models [19].. Thus, the observed significant
hypoglycemic effect of extract of V. C may be
due to the presence of these antihyperglycemic
agent and their synergistic properties.

before administration of Vernonia amygdalina

extract. Since the pancreatic cells are be
being
destroyed by alloxan, there is deficiency of
insulin leading to increased ketone body
formation. This increase in ketone bodies with
increase lipolysis leads to a severe body weight
loss [19]. However, on administration of extract
of V.C, the diabetic rats showed a significant
(P<0.05) increase and body weight gain
compared to the normal control. This could be
due to a better control of hyperglycemic state in
the diabetic rats and decreased fasting blood
glucose level could improve body weight in
alloxan-induced diabetic rats [16].. Studi
Studies have
shown an association between hyperglycemia
and decrease in body weight of experimental
animals [20].. Consequently, changes in weight
are usually a fundamental index of physiological
or pathological state of an experimental animal.

It was observed in the study that alloxan

alloxan-induced
diabetic rats had a reduction in body weight
which later appreciated during the experimental
period. This would be expected as one of the
pronounced effect of diabetes (body weight loss),
an observation which was also reported by
Atangwho et al. [16] weight loss in diabetic rats

Fig. 4. Comparison of triglyceride concentrations in the different experimental groups

Values are expressed
xpressed as mean + SEM; n = 4; No significant differences among groups

Fig. 5. Comparison of total cholesterol concentrations in the

e different experimental groups
Values
es are expressed as mean + SEM; n = 4; *significantly different from NC at p<0.05; a = p<0.05 vs diabetics;
b = p<0.05 vs insulin

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Iwara et al.; EJMP, 8(2): 78-86, 2015;; Article no.EJMP.2015.092

no.

Fig. 6. Comparison of high density lipoprotein cholesterol concentrations in the

different experimental groups
Values
ues are expressed as mean + SEM; n = 4; No significant differences among groups

Fig. 7. Comparison of very low density lipoprotein cholesterol concentrations in the different
experimental groups
Values
es are expressed as mean + SEM; n = 4; No significant differences among groups

Lipid
abnormalities
accompanying
with
premature atherosclerosis is the major cause of
cardiovascular diseases and is one of the
complication associated with diabetic patients,
therefore treatment option the management of
diabetic conditions, in addition to glycemic
control, should have a favorable effect on lipid
profile. Cardiovascular diseases are listed as the
cause of death in 65% people suffering from
diabetes [21,22]. The abnormal high level of
serum lipids is mainly due to the uninhibited
actions of lipolytic hormones on the fat depots,
mainly due to the action of insulin. Under normal
circumstances, insulin activates the enzyme
lipoprotein lipase, which hydrolyses triglycerides.

However, in diabetic state lipoprotein lipase is

not activated due to insulin deficiency, resulting
in hyper triglyceridemia [23], and insulin
deficiency is also associated with hyper
cholesterolemia due to metabolic abnormalitie
abnormalities.
The dyslipidemia is characterized by increase in
TC, LDL, VLDL, TG and fall in HDL. This altered
serum lipid profile was reversed towards normal
after treatment with extract of V.C
V.C. From this
point of view, it was encouraging that the 14 day
treatment of extract of V.C brought down the
elevated level of lipid profile such as total
cholesterol, triglycerides, HDL-cholesterol,
cholesterol, VLDL
VLDLcholesterol and LDL-cholesterol
cholesterol compared to
both the control and insulin treated groups.

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Iwara et al.; EJMP, 8(2): 78-86, 2015;; Article no.EJMP.2015.092

no.

Fig. 8. Comparison of low density lipoprotein cholesterol concentrations in the

th different
experimental groups
Values are expressed
sed as mean + SEM; n = 4; *significantly different from NC at p<0.05;
a = p<0.05 vs.
vs diabetics; b = p<0.05 vs insulin.

2.

4. CONCLUSION
Observed from this result, it may be deduced that
Vernonia calvoana may be acting in a similar
fashion to the insulin (standard drug) in reducing
the elevated blood glucose level and lipid profile
of alloxan induced diabetic rats.

3.

4.

Therefore, the results provide pharmacological

evidence for its folklore claim as an anti
anti-diabetic
and hypolipidemic agent. Further studies on its
isolation, identification
ntification and characterization of the
active principles would go a longway in giving
more light on their mechanism of action.

5.

CONSENT
It is not applicable.

ETHICAL APPROVAL

6.

It is not applicable.

COMPETING INTERESTS
Authors have
interests exist.

declared

that

no

competing
7.

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