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Alexandria Journal of Veterinary Sciences 2014, 41:19-27
ISSN 1110-2047, www.alexjvs.com
DOI: 10.5455/ajvs.154650
Influence of Combined Administration of Turmeric and Black Seed on Selected
Biochemical Parameters of Diabetic Rats
Sabry M. El-Bahr1,2*, Nabil M. Taha1, Mahdy A. Korshom1, Abd EL-Wahab A. Mandour1,
Mohamed A. Lebda1
1Department of Biochemistry, Faculty of Veterinary Medicine, Alexandria University, Egypt
2Department of Physiology, Biochemistry, and Pharmacology, Faculty of Veterinary Medicine and Animal Resources,
King Feisal University, Saudi Arabia
Key words
ABSTRACT:
Diabetes
mellitus,
blood
glucose,
body
weight,
water intake,
food intake.
The objective of the present study was to investigate the effect of Turmeric (Curcuma Longa)
and Black cumin seed (Nigella Sativa) mixture on selected biochemical parameters of
streptozotocin (STZ) induced diabetic rats. Therefore, twenty one healthy adult albino rats
divided into three groups (7 rats in each). Rats of first and second groups were non diabetic
and diabetic respectively. Rats of the third group were diabetic and received a mixture of
Turmeric (0.5 g/kg b .wt.) and Black cumin seed (1 g/kg b.wt.) orally by intragastric
intubations once a day for six weeks. The changes in blood glucose level, body weight and
water and food intake were measured. In addition, selected biochemical parameters were also
determined. The present study revealed that, dietary supplementation of plant mixture of
Turmeric and Black cumin seed to diabetic rats significantly (P<0.05) reduced the blood
glucose level as well as water and food intake accompanied by an increase in body weight
gain when compared with untreated diabetic rats. Proteins were not affected whereas liver
and kidney functions were disrupted in STZ-diabetic rats and restored whenever treated by
medicinal plants mixture. Hypercholesterolemia and hyperlipidemia observed in STZ-
diabetic rats were attenuated by administration of examined medicinal plants combination.
Conclusively, dietary supplementation of Turmeric and Black cumin seed mixture reduced
blood glucose level and ameliorated the hypercholesterolemia and hyperlipidemia in STZ
induced diabetic rats.
Corresponding Author: Sabry M. El-Bahr: e-mail: sabryelbahr@hotmail.com;
1. INTRODUCTION
Diabetes is one of the most common
chronic diseases affecting more than 100
million people worldwide. The two major
types of diabetes mellitus are characterized by
hyperglycemia, abnormal metabolism of lipid
and protein with specific long term
complications affecting the retina, kidney and
nervous system (Mathur et al., 2011).
Hyperglycemia is an important factor in the
development and progression of the
complications of diabetes mellitus (Luzi,
1998). The literature on medicinal plants with
hypoglycemic activity is vast. Many of these
plants were used for many centuries without
any side effects. However chronic
consumption of large amounts of traditional
remedies must be taken with caution as
toxicity studies have not been conducted for
most of these plants (Shnkar et al., 1980).
Turmeric (Curcuma longa) is a
perennial herb that grows to a height of three
to five feet and is cultivated extensively in
Asia (India and China) and other countries
with a tropical climate. Curcumin, is the
active ingredient of turmeric and is a potent
antioxidant and anti-inflammatory agent with
hepatoprotective, anticarcinogenic and
El-Bahr et. al. /Alexandria Journal of Veterinary Sciences 2014, 41:19-27
20
antimicrobial properties (Duvoix et al.,2005;
Aggarwal et al., 2005) . Curcumin also has a
beneficial effect on blood glucose in diabetics
and increases gastric mucosal secretion in
rabbits (Arun and Nalini 2002). The Turmeric
rhizomes have been reported to possess
antidiabetic properties as its alcohol extract
possesses active constituents showing blood
glucose lowering activity in alloxan induced
diabetic rats (Shnkar et al., 1980). Black
cumin seed (Nigella sativa) is herbaceous
plant which is a member of the Ranunculocea
family. Black seed induced significant
reduction in blood glucose and cholesterol
levels in humans (Bamosa et al., 1997). Black
cumin seed has been found to rank high
among the antidiabetic plants most
recommended by traditional practitioners
(Haddad et al., 2003). Many studies have also
examined the antidiabetic effect of Black
cumin seed (Nigella sativa) in normal and in
diabetic animal. Beside the effect of crude
aqueous extract of Black cumin seed on
glucose homeostasis (Labhal et al., 1997) its
petroleum ether extract significantly lowered
fasting plasma levels of insulin and
triglycerides and normalized high density
lipoprotein cholesterol (HDL-c) (Le et al.,
2004). Because of publications regarding the
antidiabetic effect of Turmeric and black
cumin seed mixture are scarce, the present
study was conducted to study the synergistic
efficacy of both plants as a mixture on blood
glucose level, water and food intake, body
weight and some biochemical parameters in
STZ induced diabetic rats.
2. MATERIALS AND METHODS
2.1. Preparation of plant suspension
The whole Black cumin seeds were
crushed in a blender and 12.5 g of the seeds
was added to 100 ml distilled water at room
temperature to prepare a crude suspension a
few minutes before experiment (Al-Ghamdi,
2001). Turmeric powder was mixed with
distilled water (Meddah et al., 2009; Mills and
Bone 2000). The Black cumin seed was used
in a dose of 1 g/kg body weight and mixed
with Turmeric powder in a dose of 0.5 g/kg
body weight. The dose used in the current
experiment was the half of the doses used in
previous works for Black cumin seed (Labhal
et al., 1997; Le et al., 2004; Meddah et al.,
2009) and Turmeric (Arun and Nalini 2002)
because plants were used in a combination.
2.2. Experimental animals
A total of twenty one male albino rats
(3 months old) weighing between 185–200 g
were housed in clean and disinfected cages.
Commercial basal diet and water were
provided ad libitum. Rats were subjected to
natural photoperiod of 12hr light:dark cycle
throughout the experimental period (6 weeks).
All rats received basal diet for two weeks
before the start of the experiment for
adaptation and to ensure normal growth and
behavior. Rats were maintained in their
respective groups for six weeks, monitored
closely every day and weighed every week.
The food of rats was weighed every day
before and after feeding the animals to
determine the daily food intake.
2.3. Induction of experimental diabetes
Diabetes was induced by administering
intraperitonial injection of a freshly prepared
solution of STZ (60 mg/kg b. w.) in 0.1 M
cold citrate buffer (pH 4.5) to the overnight
fasted rats (Sekar et al., 1990). After 72 hours,
fasting blood glucose levels were monitored.
Animals having blood glucose values above
250mg/dl on the third day after STZ injection
were considered as diabetic rats (Tank et al.,
1989). Then the treatment was started on the
third day after STZ injection and it was
considered as first day of treatment.
2.4. Experimental design and sampling
analysis
The rats were divided into three groups
(7 rats each).
Group 1: non diabetic control rats (negative
control).
Group 2: Diabetic control rats (positive
control).
Group 3: Diabetic rats treated with black
cumin seed (1 g/kg b.wt. per day) mixed with
turmeric (0.5 g/kg b.w. per day) in aqueous
solution orally for six weeks.
Daily measurements of body weight,
and water and food intake were recorded. For
blood glucose level estimation, blood samples
El-Bahr et. al. /Alexandria Journal of Veterinary Sciences 2014, 41:19-27
21
were collected from the fasted rats of three
groups prior to the treatment with plants and
three times per weeks after oral administration
of the treatments up to 6 weeks. The blood
samples were collected by snipping tail with
sharp razor and blood glucose level was then
measured immediately by glucose strips
(haemo-glucotest). In addition, blood samples
were collected without anticoagulant for
serum separation. Serum was separated by
centrifugation for 10 min at 3000 rpm and
was immediately frozen at –20ºC until the
time of analysis. The sera were used for
spectrophotometric determination of the
activities of Aspartate Transaminase (AST)
and Alanine Transaminase (ALT) (Reitman
and Frankel, 1957). In addition, total protein
(Doumas et al., 1981), albumin (Reinhold,
1953), globulin, uric acid (Tabacco et al.,
1979), creatinine (Henry, 1984),
triacylglycerol (TAG) (Sidney and Bernard
1973) and total cholesterol (Zak et al., 1954)
were also determined. Very low density
lipoprotein cholesterol (VLDL-c) was
calculated by division of TAG by 5 (Bauer,
1982). Calcium, phosphorus and magnesium
were determined by using commercial kits on
chemistry analyzer according to the
manufacturer instructions.
2.5. Statistical analysis
All the grouped data were statistically
evaluated and the significance of changes
caused by various treatments was determined
using Students t-test (Chou, 1975). The
results have been expressed as means ± SD
from seven rats in each group. Wherever
appropriate, one way ANOVA was also done.
The statistical significance was set at P <0.05.
3. RESULTS
The present findings (Table 1)
showed that the control rats (group I) gained
weight over the six weeks of the experimental
period, with an average body weight
increasing by 60 grams. Moreover, the
untreated diabetic rats (group II) lost an
average of 30 grams of their weight after six
weeks (P<0.05). When these diabetic rats
treated with a mixture of Black seed and
Turmeric did not lost their weight but gained
20 grams of body weight which represented
33.3% of weight gained in normal non
diabetic control rats.
Data summarized in Table 2 revealed
that, water intake in untreated diabetic groups
(Group II) were significantly (P<0.05)
increased by 100% than non-diabetic normal
rats (group I). When these diabetic rats treated
with plant mixture, 30% of the increase in
water intake in diabetic rats was decreased
(P<0.05) throughout the entire intervals of the
experiment.
Table 1 Effect of oral administration of Turmeric and Black cumin seed mixture for six weeks on body
weight gain (g) in streptozotocine diabetic rats.
Groups
Body weight (g)
Gain in body weight (g)
Initial
Sixth weeks
I
195 ± 5.1
255 ± 6.2a
60
II
194 ± 7.2
164 ± 7.1c
-30
III
194 ± 6.1
214 ± 5.2b
20
I (non diabetic), II (diabetic), III (diabetic treated with plant mixture), Values are mean ± SD of 7 rats ,
Means within the same column with different letters are significantly differed (P ≤ 0.05).
Table 2 Effect of oral administration of Turmeric and Black cumin seed mixture for six weeks on
water intake (ml/day) in streptozotocine diabetic rats.
Groups
Water intake (ml/day)
1st week
2nd week
4th week
6th week
I
50 ± 3.3c
51 ± 5.1c
55 ± 4.1c
60 ± 4.6c
II
100 ± 5.2a
95 ± 5.2a
90 ± 5.1a
87 ± 4.7a
III
70 ± 5.5b
75 ± 2.1b
75 ± 5.1b
75 ± 2.9b
I (non diabetic), II (diabetic), III (diabetic treated with plant mixture), Values are mean ± SD of 7 rats
Means within the same column with different letters are significantly differed (P ≤ 0.05).
El-Bahr et. al. /Alexandria Journal of Veterinary Sciences 2014, 41:19-27
22
Table 3. Effect of oral administration of Turmeric and Black cumin seed mixture for six weeks
on feed intake (g/day) in streptozotocine diabetic rats.
Groups
Food intake (g/day)
1st week
2nd week
4th week
6th week
I
15 ± 2.1c
17 ± 1.2c
20 ± 1.1c
22 ± 2.1c
II
38 ± 1.9a
40 ± 1.5a
39 ± 1.4a
41 ± 1.7a
III
22 ± 1.7b
26 ± 1.1b
25 ± 2.1b
30 ± 1.4b
I (non diabetic), II (diabetic), III (diabetic treated with plant mixture), Values are mean ± SD of 7 rats
Means within the same column with different letters are significantly differed (P ≤ 0.05).
Table 4 Effect of oral administration of Turmeric and Black cumin seed mixture for six weeks on
blood glucose concentration (mg/dl) in streptozotocine diabetic rats
Groups
Blood glucose concentration (mg/dl)
1st week
2nd week
4th week
6th week
I
99 ± 2.1c
100 ± 1.2c
98 ± 1.1c
96 ± 2.1c
II
350 ± 1.9a
355 ± 1.5a
360 ± 1.4a
355 ± 1.7a
III
250 ± 1.7b
240 ± 1.1b
245 ± 2.1b
240 ± 1.4b
I (non diabetic), II (diabetic), III (diabetic treated with plant mixture), Values are mean ± SD of 7 rats
Means within the same column with different letters are significantly differed (P ≤ 0.05).
Data summarized in Table 3 revealed
that feed intake in untreated diabetic groups
(Group II) were significantly (P<0.05)
increased by 153% than non-diabetic normal
rats (group I). When these diabetic rats treated
with a mixture of Black seed and Turmeric,
39.5% of the increase in feed intake in
diabetic rats was reduced (P<0.05) throughout
the entire intervals of the experiment.
The results for the effects of
Turmeric and Black cumin seed mixture on
blood glucose concentration of diabetic rats
are shown in Table 4. At the start of the
experiment, there were no significant
(P>0.05) differences in the mean values of
blood glucose level between all experimental
groups. After injection of STZ, the mean
values of blood glucose levels in untreated
diabetic rats (group II) were remained above
350 mg/dl during the entire period of the
experiment, which were significantly
(P<0.05) higher than those of the non-diabetic
normal control rats by 253.3%. The
percentage of increase in glucose level of
diabetic rats (253.3%) was reduced by 28.6%
(250 mg/dl) when they treated with plants
combination.
The results presented the effect of
Turmeric and Black cumin seed mixture on
protein patterns, lipid profile, liver and kidney
functions and electrolytes were presented in
Table 5. These findings indicated that, STZ
induced significant (P<0.05) increase in total
cholesterol values by 31.3% compared with
normal non diabetic rats. The plants
combination restored the cholesterol to
normal level. The present findings indicated
also that, STZ induced significant (P<0.05)
increase in TAG values by 9% (142 ±
3.4mg/dl) compared with normal non diabetic
rats (130 ± 4.1 mg/dl). Combined
administration of plants reduced this increase
till reached 41.5% (76 ± 4.1 mg/dl) lower than
the normal control level (130 ± 4.1 mg/dl).
The other examined biochemical values
remained unchanged throughout the
experimental period.
4. DISCUSSION
Growth promoters are commonly
added to the animals feed for growth
enhancement and efficient feed utilization.
They are chemical products, antibiotics,
enzymes and/or natural extractives. Since the
use of chemical products antibiotics might
have some unfavorable side effects, therefore
researchers tended to use natural additives
El-Bahr et. al. /Alexandria Journal of Veterinary Sciences 2014, 41:19-27
23
which meet the requirements of good growth
promoting agents (Fluk et al., 1976).
Medicinal plants have received increasing
attention as spices for human and additive in
diets for animals. In the present study, we
tried to examine the antidiabetic effect of
Black cumin seed and Turmeric mixture in
experimentally induced diabetic rats, since the
publication concerning this issue are limited.
Injection of STZ in rats caused significant
increase in blood glucose level. Using 60
mg/kg STZ dose induced an autoimmune
process that resulted in the destruction of the
ß-cells of the islets of Langerhans and the
same dose of STZ resulted in the toxicity of
ß-cells with emergence of clinical diabetes
within 2-4 days (Weiss, 1982). Previous
articles (Hussain, 2002; Sellamuth et al.,
2009) indicated that, STZ diabetic rats are
hyperglycemic and have increased oxidative
stress. This is also in consistent with the fact
that blood glucose level might have increased
due to gluconeogenesis in the absence of
insulin (Yao et al., 2006). On other hand, the
present study revealed that Turmeric and
Black cumin seed mixture was effective in
reducing blood glucose concentration. The
hypoglycemic effect of Turmeric comes in
accordance with those obtained in rats (Arun
and Nalini 2002; Sharma et al., 2006).
Authors reported that curcumin, the active
principles of Turmeric had a beneficial effect
on blood glucose in diabetics and increases
gastric mucosal secretion in rabbits. The
inclusion of curcumin at 0.6-0.9 g/kg diet
improved the adverse effect of aflatoxins in
blood glucose values of broiler chicken
(Ahmadi, 2010). The present results
concerned the hypoglycemic effect of plant
mixture agrees with those reported the
antidiabetic effect of Turmeric rhizomes in
alloxan induced diabetic rats (Shnkar et al.,
1980) and with others (Haddad et al., 2003;
Labhal et al., 1997; El-Bagir et al., 2010) that
demonstrated the hypoglycemic effect of
Black cumin seed. The antidiabetic effect of
Turmeric perhaps attributed to the antioxidant
activity of curcumin against oxidative tissue
damage (Joe and Lokesh, 1994) whereas, the
antidiabetic effect of Black cumin seed
perhaps explained by an insulin-like
stimulation of glucose uptake by muscle and
adipose tissue (Benhaddou-Andaloussi et al.,
2010) or inhibition of intestinal glucose
absorption (Meddah et al., 2009).
Table 5 Effect of oral administration of Turmeric and Black cumin seed mixture for six weeks on
selected biochemical parameters in streptozotocine diabetic rats
Parameters
Group I
Group II
Group III
Total Protein (g/l)
5.5 ± 0.2
5.4 ± 0.1
5.6 ± 0.1
Albumin (g/l)
3.2 ± 0.2
3.3 ± 0.1
3.4 ± 0.2
Globulin (g/l)
2.3 ± 0.2
2.1 ± 0.1
2.2 ± 0.1
A/G ratio
1.4 ± 0.1
1.6 ± 0.1
1. 5 ± 0.1
Total cholesterol (mg/dl)
48 ± 2.2b
63 ± 3.5a
49 ± 2.3b
TAG (mg/dl)
130 ± 4.1b
142 ± 3.4a
76 ± 4.4c
ALT (U/l)
19 ± 1.2
18 ± 1.4
20 ± 2.4
AST (U/l)
100 ± 4.5
102 ± 4.1
100 ± 4.4
BUN (mg/dl)
4.1 ± 1.6
3.8 ± 2.1
5.1 ± 2.0
Uric acid (mg/dl)
2.1 ± 0.3
2.3 ± 0.2
2.0 ± 0.1
Creatinine (mg/dl)
0.5 ± 0.1
0.6 ± 0.2
0.6 ± 0.1
Calcium
10.6 ± 2
11.5 ± 0.8
10.5 ± 1
Phosphorus
2.1 ± 0.2
4.5 ± 1.9
2.6 ± 0.8
Magnesium
1.5 ± 0.9
0.8 ± 0.2
1.0 ± 0.3
Chloride
109.6 ± 16
104.6 ± 5.5
107.2 ± 4.5
I (non diabetic), II (diabetic), III (diabetic treated with plant mixture), Values are mean ± SD of 7 rats
Means within the same column with different letters are significantly differed (P ≤ 0.05).
El-Bahr et. al. /Alexandria Journal of Veterinary Sciences 2014, 41:19-27
24
The present study also showed that,
injection of rats with STZ lead to reduction of
mean value of body weight accompanied by
an increase in the mean values of food and
water intake. These findings further support
the fact that the most classic symptoms for
diabetes mellitus are Polyuria, Polydipsia and
Polyphagia. Poor or defect of glucose
utilization can also lead to weight loss and
sense of fatigue despite of normal or even
increased of food intake (Irshaid et al., 2010).
In addition, body weight loss and fatigue can
also result due to loss of fluid. The animals
should increase food and water intake to
compensate for loss of body weight and fluid
as a consequence to such changes (Irshaid et
al., 2010). On other hand, the present data
also revealed that treatment of diabetic rats
with Turmeric and Black cumin seed mixture
appears to have positive effects on body
weight gain as well as in reduction of both
food and water intake. These ameliorative
effects might be explained by the partial
reduction in blood glucose levels in STZ
induced diabetic rats. However, another study
(Ahmadi, 2010) reported that, oral
administration of Turmeric (0.3 or 0.6 g/kg
diet) had no effect on growth performance of
broiler chicken. An increase in body weight
gain in rats fed different concentration of
Black cumin seed was reported (El-Bagir et
al., 2010) whereas in the contrary, oral
administration of methanol extracts of Black
cumin seed lowered body weight in mice
(Vahdati-Mashhadian et al., 2005).
Interestingly, the effect of combined mixture
of Black cumin seed and Turmeric was
investigated only in fish (El-Bahr and Saad
2008). The dietary supplementation of Black
cumin seed and Turmeric (5g/kg diet)
improved performance and biochemistry of
Mugil Cephalus (El-Bahr and Saad 2008).
The unchanged serum total
proteins, albumin and globulin values in STZ
diabetic rats treated with plant mixture
disagree with the findings observed in Mugil
cephalus fish (El-Bahr and Saad 2008) when
the same dose of plant mixture was used. The
present results concerning biochemistry
disagree with previous researches
demonstrating the positive effect of Black
cumin seed in biochemical parameters of Nile
tilapia (Hussain, 2002), Catfish (Ezat, 2005),
Pekin ducklings (El-Bahr, 2007) and Mugil
Cephalus fish (El-Bahr and Saad 2008) and
disagree also with the results concerning the
positive effect of Turmeric in biochemical
values of mice (Antony et al., 1999), rats
(Salama and El-Bahr 2007) and Mugil
Cephalus fish (El-Bahr and Saad 2008).
However, this difference perhaps attributed to
different species, dose and experimental
period. The increased activity of AST in STZ
induced diabetic rats indicated liver
dysfunction as reported before in rats (Saeed
et al., 2008). Liver damage was observed after
oral administration of aqueous extract of
Nigella sativa (10 ml/kg of body weight for
14 consecutive days) in rats (Tennekoon et
al., 1991) or in mice after oral administration
of an aqueous extract of the Black cumin
seeds in 4 different doses, 6, 9, 14 and 21 g/kg
(Vahdati-Mashhadian et al., 2005). In
addition the same liver damage was observed
in ducklings kept on powdered Black cumin
seed (20 gm/kg diet) (El-Bahr and Saad
2008). The liver damage observed in the
previous studies (Tennekoon et al., 1991;
Vahdati-Mashhadian et al., 2005; El-Bahr,
2007) disappeared in the present study and
that might be attributed to difference in plant
source, dosage, route of administration,
species, age and administered ingredients.
The present study reported non-significant
changes in liver and kidney function in all
groups. These results come in accordance
with the previous findings (Abubakar et al.,
2009) demonstrated that diabetes induction
caused insignificant changes in serum levels
of ALT, AST, urea and creatinine. In the
contrary, the present findings also disagree
with others (Saeed et al., 2008) demonstrated
an increase in blood urea nitrogen and uric
acid in STZ induced diabetic rats indicated
kidney dysfunction.
Diabetes induced many changes in
lipid profile as hyperlipidemia and
hypercholesterolemia observed in STZ
diabetic rats come in accordance with the
previous studies in rats (Hussain, 2002; Soni
El-Bahr et. al. /Alexandria Journal of Veterinary Sciences 2014, 41:19-27
25
and Kuttan 1992). The antihyperlipidemic and
antihypercholesterolemic effect of Turmeric
was reported before in rats (Kim and Kim
2010). Administration of 500 mg curcumin to
human volunteers for one week did not
produce significant change in serum
cholesterol level (Soni and Kuttan 1992).
Additionally, consumption of Curcumin over
1 month or 6 months was reported not to
affect blood concentrations of TG, or TC,
LDL, and HDL-c (Baum et al., 2007). The
hypocholesterolemic effect of Turmeric
reported to be due to up-regulation of
cholesterol 7α-hydroxylase in rats (Kim and
Kim 2010). Hypolipidemic and
hypocholesterolemic effect of Black cumin
seed come in accordance with previous
research in rats (Baum et al., 2007; Kocyigit
et al., 2009; Nader et al., 2010). In the
contrary, hypercholesterolemic and
hyperlipidemic effects followed oral
administration of Black cumin seed (2%) in
Pekin ducklings were recorded (El-Bahr and
Saad, 2008). The hypolipidemic effect of
Black cumin seed may be attributed to the
synergistic effect of its different constituents,
soluble fiber, sterols, flavenoids and high
content of polyunsaturated fatty acids.
Hypolipidemic effect of soluble fibers
contents of Black cumin seed might be related
to decreased cholesterol absorption and
increased bile acid synthesis and degradation
(Ali and Blunden 2003). The
hypocholesterolemic effect of black cumin
seed may relate to up-regulation of low
density lipoprotein receptors and inhibition of
3-hydroxy 3-methylglutaryl coenzyme A
reductase (Al-Naqeep et al., 2009). The
present study can concluded that, dietary
supplementation of Turmeric and Black seed
mixture to diabetic rats was effective in
reducing blood glucose, water and food
intake, TAG and Cholesterol accompanied by
an increase in body weight gain in STZ
diabetic rats.
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