Middle East Journal of Applied Volume : 07 | Issue :04 |Oct.-Dec.

| 2017
Sciences Pages: 967-973
ISSN 2077-4613
Effect of fenugreek seeds on kidney structure and some physiological parameters
in rats
Mostafa Ismail Badr

Animal Production Department, Faculty of Agriculture, Al-Azhar University, Cairo, Egypt

Received: 02 August. 2017 / Accepted: 08 Nov. 2017 / Publication date: 14 Dec. 2017

ABSTRACT

This study was undertaken in the animal house belong to Animal Production Department,
Faculty of Agriculture, Al-Azhar University. Twenty-four male albino rats weighed 100-130 g were
divided into 4 equal groups G1, control group, G2, G3 and G4 groups fed diet contained 2.5, 5 and 7.5
% fenugreek, respectively. Rats provided feed and water adlibitum. All animals were healthy and
clinically free from any diseases. Blood samples were collected after 4 and 8 weeks from the start of
the experiment. At the end of the experiment, rats were sacrificed to obtain the kidneys. The aim of
the study was to investigate the effect of using different levels of fenugreek on kidney structure and
some physiological parameters in albino rats. Results indicated that the kidney histopathological
sections of the control and 2.5% fenugreek showed normal glomeruli, tubules and cortical blood
vessels. On the other hand, renal section of groups treated with 5% or 7.5% fenugreek, were observed
mild ischemic changes for glomeruli (small sizes). Treatment of rats with 5% or 7.5% fenugreek
significantly increased serum urea and creatinine levels, while serum glucose, cholesterol and
triglycerides significantly decreased. Treatment of rats with 2.5% fenugreek for 8 weeks did not show
any significant effect on serum urea, creatinine and glucose levels, while serum cholesterol and
triglycerides significantly decreased.
It could be concluded that 2.5% fenugreek is safe to be used as a hypocholeterolemic agent
without any side effect for better kidney structure and function.

Key words: Rat, fenugreek, kidney function, cholesterol, triglycerides

Introduction

The vegetation on the earth is a perennial and renewable source of food and energy which is
necessary for the survival of most organisms. Plants are the green factors in our planet; they convert
carbon dioxide and water to carbohydrates, and nitrogen to amino acids. Besides food, plants are
considered the nature’s green pharmacy, which provide drugs to maintain the good health and to
restore the failed health of humans. The medical arts had its origin when mankind first began to use
remedial measures to get rid of their pains, sufferings and other illnesses (Badr et al., 2012).
Trigonella Foenum graecum L. (fenugreek) is a legume originated in south Eastern Europe and
Western Asia, andgrow now in India as well as in certain parts of Asia, Northern Africa, Europe and
the United States (Altuntas et al., 2005). Fenugreek is one of the most widely used medicinal plants in
folk medicine (Subramanian and Prasath 2014).Fenugreek seeds are traditionally used for treatment of
many diseases. Studies showed that plant seeds have antioxidant properties. Many medicinal
properties are attributed to fenugreek seed and leaves (Abduljawd, 2015 and Srinivasan,
2006).Fenugreek seeds are commonly used as condiment and seasoning in food preparation (Renuka
et al., 2009).Fenugreek abundant in polyphenolic compounds (Rayyan et al., 2010) and known to
have several beneficial action, such as antioxidant effect (Ravikumar and Amerada, 1999), anti-
diabetic effects (Haeri et al., 2009) and hypocholesterolemic effect (Sowmya and Rajyalakshmi,
1999).
The objective of this study was to assess the effect of supplementing rats with different levels
of Trigonella foenum graecum seeds on kidney structures and some physiological parameters of male
albino rats.

Corresponding Author: Mostafa Ismail Badr, Animal Production Department, Faculty of Agriculture, Al-
Azhar University, Cairo, Egypt. E-mail: dr.mostafaismail77@yahoo.com
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Materials and methods

The experiments were carried out in the animal house of Animal Production Department,
Faculty of Agriculture, Al-Azhar University, Cairo, Egypt.

Experimental Animals:

Twenty-four male albino rats ranged 100-130gm weight were obtained from El-Osman farm,
Cairo, Egypt .Animals housed in stainless steel cages and provided with feed and water ad libitum.
All animals were healthy and clinically free from diseases.

Experimental Design:

Seeds of Trigonella Foenum graecum (fenugreek) were used in the present study. Fenugreek
were obtained from the local market in Cairo.
Animals were divided randomly into 4 equal groups, each of 6 rats. G1, control group, G2, G3
and G4 groups fed diets contained 2.5, 5 and 7.5 % fenugreek, respectively. Rats treated with the
experimental diets for 8 weeks. During the experiment, blood samples were collected 4 and 8 weeks
after beginning of the experiment. Blood samples were obtained from all rats by withdrawing blood
from the orbital venous plexuses using a capillary tube. Blood were centrifuged at 3000 rpm for 15
min to obtain serum which transferred to Ependroff tubes and stored at -20oC until subsequent
analyses. At the end of experiment, rats were sacrificed to obtain their kidneys. Immediately after
extraction, the kidneys were immersed in formalin 10% for two days, washed in water, dehydrate in
ascending grade of ethyl alcohol and finally cleared by xylene and embedded in melted paraffin wax.
The kidney blocks were sectioned at six–micron thickness and stained by eosin and heamatoxylin
according to Pearse, (1968).

Serum parameter:

Serum urea measured according to the colorimetric method described by Tabacco et al.
(1979). Serum creatinine concentration was determined by using enzymatic colorimetric method
according to Junge et al. (2004). Serum glucose measured according to the oxidase method (Trinder
1969). Serum cholesterol measured by enzymatic colorimetric method based on the method of
Roeschlau et al. (1974). Serum triglycerides were determined using enzymatic colorimetric method
according to Bucolo and David (1973).

Statistical Analysis:

Statistical analysis was conducted by analysis of variance using SPSS Package (2016). The
means and standard errors of all parameters were calculate and Duncan Multiple Range test (Duncan,
1955) was used when differences were significant to explore which means are different.

Results and Discussion

Histopathological Changes of the Kidney

The Kidney structure of the control and 2.5% fenugreek group showed preserved architecture.
The cortex revealed normal glomeruli, tubules and cortical blood vessels. The medulla revealed
normal tubules and normal blood vessels of interstitial tissue (Figs 1 and 2). Kidney feature in 5% and
7.5% fenugreek groups showed preserved architecture. The cortex revealed mild ischemic changes
with small sizes glomeruli and normal tubules with normal cortical blood vessels. The medulla
revealed normal tubules and normal interstitial tissue (mild ischemic changes of glomeruli) (Figs 2
and 3).
Sayed et al. (2012) reported that renal histology of diabetic rats showed a mild diffuse
mesangial expansion compared with normal glomeruli, while rats supplemented with 5% Trigonella

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foenum graecum (fenugreek seeds powder)with rat food showed reduced in score of mesangial
expansion. They demonstrated that fenugreek seeds powder significantly improved glucose
homeostasis and renal function in alloxan diabetic rats by delaying carbohydrate digestion and
absorption, attenuating inflammation and improving kidney antioxidant status.
The present study revealed that 5 and 7.5 %fenugreek have a side effect on kidney structure as
it caused mild ischemic changes of glomeruli.
Belaid Nouira et al. (2013) showed that Alcl3 affects the renal tubule histology by changing the
aspect of their lining epithelial. The proximal convoluted tubules seemed to be the most affected.
They also showed that Alcl3 altered the apical differentiation of the cortex proximal tubules cells by
reducing the brush border density, leading to a reduction of the absorption surface area. Fenugreek
seeds powder act as a kidney protector, it succeeded to counter balance the weight loss of kidneys.
Moreover, it significantly improved the biochemical and histological parameters.
In the present study the 2.5% fenugreek group did not have any significant effect on kidney
structure, while 5 and 7.5 % fenugreek had a side effect on kidney. Fenugreek(5 or 7.5%) caused mild
ischemic changes of glomeruli. This conflict between present result and others might be attributed to
the different dose of plant, plant preparation, route of administration and time of the experiment.

Fig. 1: A section in the kidney tissue of control group showed preserved architecture. The cortex
revealed normal glomeruli, tubules and cortical blood vessels. The medulla revealed normal
tubules and normal blood vessels of interstitial tissue. (H&E., stain, x100).

Fig. 2: Shows section in the kidney tissue of 2.5% fenugreek group showed preserved architecture.
The cortex revealed normal glomeruli and tubules with normal cortical blood vessels. The
medulla revealed normal tubules and normal blood vessels of interstitial tissue. (H&E., stain,
x100).

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Fig. 3 : Shows section in the kidney tissue of 5% fenugreek group showed preserved architecture. The
cortex revealed mild ischemic changes glomeruli (small sizes) and normal tubules with
normal cortical blood vessels. The medulla revealed normal tubules and normal interstitial
tissue. (H&E., stain, x100).(Mild ischemic changes of glomeruli).

Fig. 4: A section in kidney tissue of 7.5% fenugreek group showed preserved architecture. The cortex
revealed mild ischemic changes glomeruli (small sizes) and normal tubules with normal
cortical blood vessels. The medulla revealed normal tubules and normal interstitial tissue.
(H&E., stain, x100).).(Mild ischemic changes of glomeruli).

Serum parameters

Table (1) shows that after 4 weeks from the start of experiment all doses of fenugreek seeds
(2.5, 5 and 7.5%) did not show any significant effect on serum urea compared with control group.
While after 8 weeks from the start,5% and 7.5% fenugreek significantly increased serum urea
compared with the control and 2.5% fenugreek groups, yet the low fenugreek level (2.5%) continued
having no effect on serum urea level. These results indicate that serum urea was significantly
increased by increasing the dose of fenugreek and by increase the time of treatment.

Table 1: Serum urea concentration as affected by different doses of fenugreek seeds (mg/dL)
4 weeks 8 weeks
Control 83.96 ± 4.59 A 85.49± 3.41 B
2.5% fenugreek 81.11 ± 3.47 A 86.38 ± 3.62B
5% fenugreek 82.22 ± 4.34 A 95.94 ± 4.71 A
7.5% fenugreek 84.03 ± 3.80 A 96.16 ± 3.67 A
S.E. :Standard error.
Means in columns with different superscripts differ at (P<0.05).

Belaïd-Nouira et al. (2013) reported that treatment of rats with aluminum chloride (AlCl3)
caused a significant increase in plasma creatinine level, while urea level significantly decreased
compared with control. However, treatment with fenugreek seed powder (5%) (FSP) alone did not

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cause any significant change on plasma creatinine and urea. Treatment with both maintained the
plasma creatinine and urea close to normal values.
Table (2) shows that after 4 weeks of the start experiment all levels of fenugreek did not show
any significant effect on serum creatinine compared with control group. While after 8 weeks 5and
7.5% levels of fenugreek significantly increased serum creatinine compared to either control group or
2.5 % fenugreek. These results indicate that fenugreek has a side effect on kidney functions when we
increased the dose and increased the time of treatment.
In this study, we found that treatment of rats with 5% or 7.5% fenugreek caused mild ischemic
changes of glomeruli and this effect may be the cause of the increase in serum concentrations of urea
and creatinine.

Table 2: Serum creatinine concentrations as affected by different doses of fenugreek seeds (mg/dL)
4 weeks 8 weeks
Control 3.603 ± 0.96 A 2.72 ± 0.17 B
2.5% Fenugreek 2.88 ± 0.85 A 2.243 ± 0.51 B
5% Fenugreek 2.803 ± 0.71 A 3.663 ± 0.42 A
7.5% Fenugreek 3.267 ± 0.81 A 3.78 ± 0.75 A
S.E.:Standard error
Means in columns with different Superscripts differ at (P<0.05)

Table (3) shows that after 4 or 8 weeks from the start of the experiment, 5% and 7.5%
fenugreek treatments significantly decreased serum glucose level compared with the control group.
While, 2.5% fenugreek for 8 weeks did not show any significant effect on serum glucose as compared
with the control group .

Table 3: Serum glucose concentration (mg/d1) due to the effect of different doses of fenugreek seeds.
4 weeks 8 weeks
Control 107.86 ± 2.25 A 115.66 ± 3.48 A
2.5% Fenugreek 103.72 ± 3.49 A 111.67 ± 2.76 A
5% Fenugreek 95.39 ± 1.06 B 94.67 ± 1.20 B
7.5% Fenugreek 92.14 ± 3.78 B 93.67 ± 1.45 B
S.E.: Standard error
Means in columns with different Superscripts differ at (P<0.05)

These results indicate that fenugreek has a hypoglycemic effect on normal rats and this effect
depend on the dose level of fenugreek. The noticed hypoglycemic effect of fenugreek is in agreement
with those found by Abdelatif et al. (2012).The hypoglycemic effect of fenugreek maybe due to the
effect of its content of pectin which delay the stomach empting and so delays glucose absorption
from intestine (Zeweil et al., 2015). Ali et al. (1995) revealed that the hypoglycemic effect of
fenugreek largely due to its high content of soluble fiber, which acts to decrease the rate of gastric
emptying thereby delay absorption of glucose from small intestine. The hypoglycemic effect of
fenugreek also may due to that fenugreek stimulate cells of islets of langerhans. Abdelatif et al.
(2012) showed that supplementation with fenugreek was associated with slight increase in insulin
concentration of diabetic rabbits.
Table (4) shows that treatment of rats with fenugreek (2.5, 5 or 7.5%) for 4 or 8 weeks
significantly decreased serum cholesterol compared with control group.
These results are in agreement with those found by Elmahdi and El- Bahr (2015)who reported
that the increase in total cholesterol in rats fed high cholesterol diet was modified and backed to
normal values with inclusion of fenugreek at rates 5 and 10%.
Table 4: Serum cholesterol concentrations (mg/d1) as affected by different doses of fenugreek seeds.
4 weeks 8 weeks
Control 95.69 ± 11.32 A 94.05 ± 10.90 A
2.5% Fenugreek 68.28 ± 10.76 B 67.41 ± 8.40 B
5% Fenugreek 66.04 ± 3.45 B 65.88 ± 5.87 B
7.5% Fenugreek 67.94 ± 4.78 B 66.35 ± 7.67 B
S.E.: Standard error
Means in columns with different Superscripts differ at (P<0.05)

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The hypocholesteremic effect of fenugreek seed may be due to its active ingredients such as
saponins, hemicelluloses, mucilage, tannin and pectin and these compounds help in lowering blood
LDL – Cholesterol levels by inhabiting bile salts
Table (5) shows that treatment of rats with 5 or 7.5% fenugreek for 4 or 8 weeks significantly
decreased serum triglycerides compared to control group. While, treatment with 2.5% fenugreek for 4
weeks did not show any significant effect on serum triglycerides, while after 8 week it significantly
decreased serum triglycerides. These results indicate that the effect of fenugreek on decreasing serum
triglycerides depend on the dose of fenugreek and time extend of the experiment.
The significant effect of fenugreek is in accordance with Elmahdi and El-Bahr (2015) result
which showed that supplementing rats fed high cholesterol diet (5 and 10%) with fenugreek did not
increase serum triglycerides.
The insignificant effect of fenugreek are in accordance with Elmahdi and El-Bahr (2015)
inclusion of fenugreek on the high cholesterol diets (5 and 10%) did not affect serum triglycerides
level compared to the rats fed high cholesterol diet and toward the normal control values .
Al-Hussary (1993) also reported that in normal rabbits, oral administration of a decoction of
fenugreek seeds (200mg/Kg body weight) for 5successive days had no significant effect on
triglycerides concentrations. Petit et al. (1995) showed also that in normal and diabetic rats, fenugreek
seed extract containing steroid saponins did not cause any changes in triglycerides.
On the other hand, the significant decrease of serum triglycerides due to treatment with
fenugreek is in accordance with those found by Badr (2006) that in summer rats fed normal protein
and energy diet and treated with fenugreek significantly had decrease in plasma triglyceride levels
compared with control group.
Zeweil et al. (2015) reported that treatment of rabbits with fenugreek showed a reduction in
serum triglycerides.

Table 5: Serum triglycerides concentrations (mg/d1) asaffected by different doses of fenugreek seeds.
4 weeks 8 weeks
Control 171.83 ± 4.25 A 182.300 ± 5.77 A
2.5% Fenugreek 177.66 ± 10.76A 152.83 ± 3.74 B
5% Fenugreek 132.63 ± 14.99 B 134.20 ± 7.32 C
7.5% Fenugreek 128.25 ± 6.35 B 131.67 ± 6.17 C
S.E.: Standard error
Means in columns with different Superscripts differ at (P<0.05)

Conclusion

Trigonella foenum graecum L. seeds is safe to be used at a dose of 2.5% as an effective remedy
for hypocholesterolemic in rats without any adverse side effects on kidney structure and function.

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