Journal of Ethnopharmacology 88 (2003) 7377

Effect of soluble dietary bre fraction of Trigonella foenum

graecum on glycemic, insulinemic, lipidemic and platelet
aggregation status of Type 2 diabetic model rats
J.M.A. Hannan
a
, B. Rokeya
a
, O. Faruque
a
, N. Nahar
b
,
M. Mosihuzzaman
b
, A.K. Azad Khan
a
, L. Ali
a,
a
Biomedical Research Group, Bangladesh Institute of Research & Rehabilitation in Diabetes,
Endocrine & Metabolic Disorders (BIRDEM), 122, Kazi Nazrul Islam Avenue, Dhaka 1000, Bangladesh
b
Department of Chemistry, University of Dhaka, Dhaka 1000, Bangladesh
Accepted 7 May 2003
Abstract
The soluble dietary bre (SDF) fraction of Trigonella foenum graecum (Tf-sdf) has previously been shown to reduce postprandial elevation
in blood glucose level of Type 2 model diabetic rats by delaying the digestion of sucrose. The Tf-sdf has now been investigated for its chronic
effect on serum fructosamine, insulin and lipid levels, and on platelet aggregation in Type 2 diabetic rats. Tf-sdf was administered orally twice
daily at a dose of 0.5 g kg
1
for 28 days. It lowered the serum fructosamine level (P < 0.05) with no signicant change in the insulin level
as compared with the control. Atherogenic lipids, i.e. triglycerides, cholesterol and LDL-cholesterol were found to decrease signicantly
in Tf-sdf fed rats (P < 0.01). HDL-cholesterol showed an opposite trend (P = 0.024), but serum non-esteried fatty acid (NEFA) values
paralleled the atherogenic lipids (P = 0.001). No signicant effect on platelet aggregation (%) was found although there was a tendency to
lower the aggregation (P = 0.069). It is concluded that Tf-sdf has a benecial effect on dyslipidemia and has a tendency to inhibit platelet
aggregation in Type 2 model diabetic rats.
2003 Elsevier Ireland Ltd. All rights reserved.
Keywords: Trigonella foenum graecum; Soluble dietary bre; Diabetes mellitus; Oral hypoglycemic agent; Fructosamine; Insulin; Hypolipidemic effect;
Platelet aggregation; Streptozotocin diabetic rats
1. Introduction
The seeds of Trigonella foenum graecum are reported
to possess hypoglycemic properties in both animal (Shani
et al., 1974; Ribes et al., 1984; Ajabnoor and Tilmisany,
1988; Amin et al., 1988) and human subjects (Madar
et al., 1988; Madar and Arad, 1989; Sharma et al., 1990). We
have previously shown that the whole powder, its methanol
extract, and the residue remaining after methanol extraction
have signicant hypoglycemic effects when fed simulta-
neously with glucose. The water extract of the methanol
extractive-free residue of the seed powder, which contains
almost exclusively the soluble dietary fraction of Trigonella
foenum graecum (Tf-sdf), showed signicant hypoglycemic
activity at different prandial states (Ali et al., 1995a,b).
Tf-sdf was found to be effective in Type 2 as well as in Type

Corresponding author. Tel.: +880-2-8611138/8617130/8613700;

fax: +880-2-8613004.
E-mail addresses: lali@dab-bd.org, lali@citechco.net (L. Ali).
1 (where insulin secretory capacity is almost absent) model
rats. It showed no effect on the fasting blood glucose levels
of nondiabetic and diabetic model rats. However, it showed
a signicant hypoglycemic effect (P < 0.05) in Type 2
model rats when fed simultaneously with glucose (Ali
et al., 1995a,b). We have also shown that its hypoglycemic
activity may be related to lowering of carbohydrate absorp-
tion through inhibition of disaccharidase enzyme in the gut
(Faruque et al., 1998). In the present study, we investigated
the effect of Tf-sdf on serum fructosamine, insulin and lipid
levels, and on platelet aggregation in the Type 2 rats after
chronic feeding.
2. Materials and methods
2.1. Plant materials and preparation of test sample
Seeds of the plant were purchased from local mar-
ket and botanically authenticated and voucher specimens
0378-8741/$ see front matter 2003 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/S0378-8741(03)00190-9
74 J.M.A. Hannan et al. / Journal of Ethnopharmacology 88 (2003) 7377
were deposited in the National Herbarium, Bangla-
desh.
Trigonella foenum graecum seeds were dried at 40

C
and nely powdered. The powder (300 g) was extracted
with aqueous 80% ethanol (3 400 ml, 30 min each time)
by reuxing and the aqueous 80% ethanol extract was dis-
carded. The remaining residue (100 g) was further extracted
with chloroform (3 750 ml, 30 min each time) at room
temperature. From the remaining residue, a soluble dietary
bre (SDF) was prepared (25.2 g) by the Theander and
Westerlund method (Theander and Westerlund, 1986).
2.2. Experimental animals
LongEvans male rats (weighing 180200 g) bred at
BIRDEM laboratory were used. Type 2 rat model were
developed by intra-peritoneal injection of Streptozotocin
(90 mg kg
1
body weight, dissolved in 0.1 citrate buffer,
pH 4.5) to 4872-h-old pups (Bonner-Weir et al., 1981).
Ten rats (n = 10) were fed with Tf-sdf (0.5 g kg
1
body
weight) twice daily for 4 weeks and another 10 were used
as control. Values of serum fructosamine, insulin and lipids
of control and treated groups at 0 day were matched.
2.3. Blood collection
Blood was collected from the abdominal aorta under pen-
tobarbital anesthesia. Serum and plasma were separated and
stored at 20

C until the estimation of fructosamine, in-

sulin and lipids.
2.4. Measurement of platelet aggregation
Blood samples were drawn from abdominal aorta after
pentobarbital anesthesia. The anticoagulant used was sodium
citrate (38 g l
1
, 1 vol. anticoagulant for 9 vol. of blood).
Platelet-rich plasma (PRP) and platelet-poor plasma (PPP)
were prepared by centrifugation. Platelet aggregation kinet-
ics in 200 ul of PRP was measured by optical aggregometry
using two different doses of ADP (12 and 14 M) as in-
ducer. Platelet aggregation was expressed as percentage of
the PPP transmission value.
2.5. Biochemical analysis
Serum fructosamine levels were measured by using spec-
trophotometric method, serum insulin by Rat insulin ELISA
kit (Crystal Chem Inc., USA) and lipids (TG, total choles-
terol, HDL-cholesterol, LDL-cholesterol and NEFA) were
measured in plasma by enzymatic-colorimetric method.
2.6. Statistical analysis
All analyses were done using the Statistical Package for
Social Sciences (SPSS) for Windows 7.5. Results are ex-
pressed as Mean S.D. Unpaired Student t-test was per-
formed for statistical comparison.
3. Results
3.1. Serum fructosamine
Fructosamine level was signicantly lowered in Tf-sdf
group (mmol l
1
, control versus Tf-sdf, 91.087.20 versus
71.99 6.35; P < 0.05) (Fig. 1a).
3.2. Serum insulin
Serum insulin did not differ signicantly between the two
groups (ng ml
1
, 166.09 9.02 in control versus 149.83
17.03 in Tf-sdf) (Fig. 1b).
3.3. Plasma lipids
Atherogenic lipids were lowered signicantly compared
to control (mg dl
1
, M S.D., control versus Tf-sdf; TG:
70.7817.45 versus 30.898.72, P = 0.001; total choles-
terol: mg dl
1
, 66.344.16 versus 52.225.81, P = 0.001;
LDL-cholesterol: mg dl
1
, 16.40 7.4 versus 6.12 6.9,
P = 0.014) (Fig. 2ac). HDL-cholesterol level (mg dl
1
)
increased from 35.78 5.31 to 41.89 5.11 (P = 0.024)
F
r
u
c
t
o
s
a
m
i
n
e

(
m
m
o
l
/
l
)
0
20
40
60
80
100
120
Control Tf-sdf
S
e
r
u
m

i
n
s
u
l
i
n

(
n
g
/
m
l
)
0.0
0.1
0.2
0.3
0.4
0.5
0.6
*
(a)
(b)
Fig. 1. Effect of Tf-sdf on serum fructosamine and serum insulin levels.
Values are mean S.D. (n = 10).

P < 0.05.
J.M.A. Hannan et al. / Journal of Ethnopharmacology 88 (2003) 7377 75
Control Tf-sdf
T
o
t
a
l
C
h
o
l
e
s
t
e
r
o
l
(
m
g
/
d
l
)
0
20
40
60
80
Control Tf-sdf
T
G
(
m
g
/
d
l
)
0
20
40
60
80
100
Control Tf-sdf
L
D
L
-
c
h
o
l
e
s
t
e
r
o
l
(
m
g
/
d
l
)
0
5
10
15
20
25
Control Tf-sdf
H
D
L
-
c
h
o
l
e
s
t
e
r
o
l
0
10
20
30
40
50
*
*
**
**
(a)
(c) (d)
(b)
Fig. 2. Effect of Tf-sdf on lipid proles. Values are mean S.D. (n = 10).

P < 0.05,

P < 0.01.
Control Tf-sdf
N
E
F
A
(
m
m
o
l
/
l
)
0.0
0.1
0.2
0.3
0.4
0.5
**
Fig. 3. Effect of Tf-sdf on NEFA level. Values are mean S.D. (n = 10).

P < 0.01.
(Fig. 2d). NEFA values (mmol l
1
) were signicantly low-
ered (0.32 0.11 versus 0.14 0.05, P = 0.001) (Fig. 3).
3.4. Platelet aggregation
Tf-sdf fraction resulted in a lowering tendency of platelet
aggregation in Type 2 rats (%, control versus Tf-sdf, ADP
(12 M): 55.96 10.75 versus 44.17 11.11, P = 0.078;
ADP (14 M): 57.29 14.04 versus 44.64 8.28, P =
0.069) (Fig. 4).
4. Discussion
We have shown earlier that Tf-sdf fraction can effec-
tively reduce postprandial rise of glucose in rats (Ali et al.,
1995a,b). The serum fructosamine data in the present
76 J.M.A. Hannan et al. / Journal of Ethnopharmacology 88 (2003) 7377
ADP concentration
12 micro-M 14 micro-M
P
l
a
t
e
l
e
t
a
g
g
r
e
g
a
t
i
o
n
(
%
)
0
20
40
60
80
Control
Tf-sdf
Fig. 4. Effect of Tf-sdf on platelet aggregation. Values are mean S.D.
(n = 10).
study reects the glycemic status of the rats for the last
23 weeks and it demonstrates, more conclusively, the
anti-hyperglycemic properties of the Tf-sdf fraction. It has
also been shown earlier that Tf-sdf fraction does not stimu-
late insulin secretion during a single feeding (Rokeya et al.,
1998), this has been further conrmed in this study at a
chronic feeding state. Findings from the present as well as
previous studies, therefore, suggest that the hypoglycemic
activity of Tf-sdf is not related to stimulation of insulin se-
cretion. Thus, this material may lead to postprandial glucose
reduction without any extra load on the pancreatic B cells.
High levels of total cholesterol and, more importantly,
LDL-cholesterol in blood are major coronary risk factors
(National Cholesterol Education Program Expert Panel,
1994). Administration of Tf-sdf fraction lowered both total
cholesterol and LDL-cholesterol in Type 2 diabetic rats.
Several studies show that an increase in HDL-cholesterol
is associated with a decrease in coronary risk and most
of the drugs that decrease total cholesterol also decrease
HDL-cholesterol (Wilson, 1990). In the present study,
the Tf-sdf, while lowering LDL-cholesterol, increased
the HDL-cholesterol fraction signicantly. Recent studies
show that triglycerides are also independent risk factors
for coronary heart disease (Bainton et al., 1992; National
Cholesterol Education Program Expert Panel, 1994) and in
the present study the agent decreased the triglyceride levels
signicantly. Tf-sdf fraction may reduce the triglycerides
by decreasing the non-esteried fatty acids (NEFA) in
Type 2 rats. NEFA may inuence platelet aggregation and
vascular changes by accelerating the rate of prostacyclin
in plasma (Reinila, 1981; Gjesdal et al., 1976). The plant
fraction showed a tendency to lower the platelet aggrega-
tion that might be due to the reduction of NEFA. Hence,
the resulting benet of the agent not only helps to combat
hyperglycemia, but also helps to prevent dyslipidemiaan
important risk factor for the micro- and macro-vascular
complications of diabetes.
It has been previously shown by us that Tf-sdf frac-
tion effectively inhibits carbohydrate absorption in the gut.
Therefore, the hypolipidemic action of the fraction could be
the result of retardation of carbohydrate and fat absorption
due to the presence of bioactive bre in the agent.
In conclusion, Tf-sdf has got promising antihyper-
glycemic or hypolipidemic effects and it has also a tendency
to reduce platelet aggregation in diabetic rats.
Acknowledgements
We gratefully acknowledge the nancial support of In-
ternational Program for the Chemical Sciences (IPICS),
International Foundation of Sciences (IFS) and Diabetic
Association of Bangladesh in conducting this study. We
are grateful to Prof. Salar Khan, National Herbarium,
Bangladesh for the identication of the plant.
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