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The Egyptian Journal of Hospital Medicine (January 2019) Vol. 74 (7), Page 1616-1620
31616
Received:3/11/2018
Accepted:22/11/2018
Effect of Phytoestrogen (Fennel) on Some Sex Hormones and Other
Physiological Parameters in Male Albino Rats
Eman G.E. Helal1, Nora Abdul-Aziz AL Jalaud2, Mohamed A. El-Aleem3,
Shaimaa S. Ahmed1 Bin Imam Abdulrahman ,
1
, EgyptairoC Girls),University ( Azhar-Zoology Department, Faculty of Science, Al 3
Azhar University (Boys), Cairo, Egypt-, Physiology Department, Faculty of Medicine, Al
2
University aisalF *Corresponding Author: Eman G.E. Helal, E-mail: emanhelal@hotmail.com, mobile: 00201001025364,
Orchid.org/0000-0003-0527-7028
ABSTRACT
Background: Phytoesrogenis plant derived compounds which have an estrogenic effect.
Objectives: The present study was carried out to investigate some pharmacological and biochemical effects of
fennel oil on male albino rats.
Materials and methods: Twelve animals were divided randomly into two groups. Group A: Control.
Group B: Treated rats. The rats were given an oral dose of 1 ml/kg body weight/day fennel oil once daily for one
month. At the end of the experiment, blood samples were collected for biochemical analysis.
Results: The fennel oil induced highly significant decrease (p<0.01) in total cholesterol(TC), triglycerides (TG),
low-density lipoprotein (LDL), very low-density lipoprotein (VLDL), LDL/high-density lipoprotein (HDL),
while significant increased (p<0.05) HDL. In addition, it showed highly significant decrease (p<0.02) in FSH,
testosterone and sperm count compared to normal control group.
Conclusion: Fennel herb has several therapeutic benefits, but it also has negative effect on male sex hormone
and sperm count. So men must reduce their usage of it.
Keywords: Fennel, Lipid profile, Albino rats, Physiological parameters.
INTRODUCTION
Fennel (Foeniculumvulgare) is a well-known
Mediterranean aromatic plant. It is a herb consumed
orally and having several therapeutic applications.
Besides,it is used in traditional medicine and as a
spice. It has diuretic, analgesic, antipyretic, and
antioxidant activity (1).
Its therapeutic properties are due to volatile
compounds; phenols, and flavonoids in the plant
extract. According to phytochemical studies, over 30
various terpenes and terpenoids have been identified
in fennel (2).
Moreover, other compounds such as estragole
(methyl chavicol), fenchone, trans-anethole, and α-
phellandrene are the main constituents of the fennel
seed (3).
The presence of flavonoids, cumarin, and
anethole has been also identified. Anethole and
fenchone are the main components (4,5). Cumarin has
anti-aromatase, estrogenic, reductase inhibiting, and
anti-androgen activities. Moreover, flavonoids and
anethole are also estrogenic compounds that could
depress serum levels of testosterone, and as a result,
reduce sperm production (6).
In the present study, we aim to clarify the effect
of fennel as phytoestrogen on some sexual hormone
and some physiological parameters in male albino
rats.
MATERIALS AND METHODS
Fennel oil was purchased from Cap Pharm for
Extracting Natural Oils & Herbs, Cairo, Egypt. The
oil of fennel contains 50% of trans-anethole and 25%
of fenchone.
Experimental animals
The experiment was carried out on 12 male
albino rats of the Rattus rattus strain weighting (130-
140gm) obtained from animal farm of El-Nile
Company for Pharmaceutical Product (El-Nile, Cairo,
Egypt). Animals were housed in metallic cages and
maintained under standard condition of temperature,
humidity and natural light/dark cycle along the
experimental period.
Food and water were available throughout the
experiment ad libitum.
Rats were left to acclimatize for one week before
starting the experiment.
Effect of Phytoestrogen (Fennel) on Some Sex Hormones…
1617
Experimental design
In current study, 12 male Albino rats were divided into
two equal groups (6 rats in each group) as the
following:
Group I: (control group): comprised of normal rats
and maintained on standard pellet diet and tap water
ad libitum for 30 days.
Group II: rats received orally fennel seed oil (1 ml/kg
body weight) for 30 days.
Body weight measurement
Body weight was recorded before and after the
experiment.
Blood sample collection
At the end of the experimental period, the blood
samples were collected from the retro-orbital sinus of
anesthetized rats (by ether) after overnight fasting.
Serum was separated by centrifugation at 2500 g for
15 minutes at room temperature to estimate
biochemical parameters.
Biochemical analysis
Assessment of biochemical parameters:
In the present study, total protein (TP) and
albumin concentration were estimated, then serum
globulin concentrations were calculated according to
the formula: Globulin (g/dl) = total protein (g/dl) –
albumin (g/dl).Aspartate aminotransferase (ASAT),
alanine aminotransferase (ALAT) activities,
creatinine, BUN concentrations as well as lipid profile
[total cholesterol, triglycerides and high-density
lipoprotein cholesterol (HDL-C)] were also
determined. All parameters were estimated using Bio
Merieux SA kits, France.
The ratio of serum albumin/ globulin was
determined. However, ratios of TC/HDL (risk factor
1) and LDL/HDL (risk factor 2) were also calculated
after calculation of serum LDL-C (low-density
lipoprotein cholesterol) and VLDL (very low density
lipoprotein cholesterol) using the Friedwald's(7) and
Norbert (8)formulas, respectively as following:
Friedewald's equation:
LDL (mg/dl) = TC- {HDL + [TG/5]}. Norbert
equation: VLDL = TG/5.
Hormonal assay
Estimation of serum luteinizing hormone (LH)
follicles-stimulating hormone (FSH) and testosterone
(T) levels.
They were estimated by using manufacture
instructions of kit. All kits used for hormone assay
were from Monobind Inc. lake forest CA 92630, USA.
Sperm collection and evaluation
The rats were sacrificed, and the left caudal
epididymis was separated and the total recovered
sperm volume during 4 h of incubation in normal
saline (volume=1 ml, 35∼37oC) was calculated. The
sperm concentration was determined by the
conventional method using a hemocytometer chamber
for the red blood cell count. The right epididymis was
finely minced by anatomical scissors in 1 ml of
warmed isotonic saline in a petridish.
The sperm progressive motility (SPM) was
estimated by evaluating 4 fields of asperm droplet
under a cover-slip on a warm glass slide (35∼37oC)
under light microscopy (×40). The sperm vitality was
assayed using a conventional procedure of eosin B-
nigrosin stain (1.67% eosin, 10% nigrosin, and 0.1 M
sodium citrate) under ×100 magnification and 100
sperm were counted. All of the sperm evaluation
procedures were carried out based on the World
Health Organization manual for human sperm
analysis with some modifications(9).
Ethical approval
The study was approved by the Ethics Board of Al-
Azhar University.
Statistical analysis
The results were expressed as Mean ± SEM. Data
were analyzed by independent t-test and were
performed using the Statistical Package (SPSS)
program, version 19.
RESULTS
Liver functions
Results of the present study showed non-
significant change in ALAT and ASAT in the treated
groups when compared to control rats (Table 1).
Table 1: Changes in the ALAT and ASAT levels in
the control and treated groups
Groups
Parameters
Control
Fennel oil
ALAT(U/L)
% of change
22.5±0.6
18.5±0.42
-17.7%
ASAT(U/L)
% of change
33.5±0.5
30.5±0.56
-8.9 %
Values represent mean ±SE (stander error).
Lipid profile
Fennel oil induced highly significant decrease
(p<0.01) in TC, TG, LDL, VLDL, LDL/HDL. And
significant increase (p<0.05) in HDL, as compared to
control animals (Table 2).
Eman Helal et al.
1618
Table 2: Changes in the lipid profile levels in the control and treated groups
Groups
Parameters
Control
Fennel oil
TC(mg/dl)
% of change
141.3±0.42
95.0±0.0001**
-32.7%
TG(mg/dl)
% of change
71.5±1.56
43.0±0.0001**
-39.8%
HDL(mg/dl)
% of change
50.33±0.33
62.7±0.0001**
-24.5%
LDL(mg/dl)
% of change
71.2±1.27
23.7±0.0001**
-66.7%
VLDL(mg/dl)
% of change
14.30±0.31
8.6±0.0001**
-39.8%
LDL/HDL(mg/dl)
% of change
1.22±0.007
0.37±0.0001**
-69.6%
TC/HDL(mg/dl)
% of change
2.47±0.008
1.51±0.001**
-38.8%
Values represent mean ±SE (stander error). (p*<0.05, p**<0.001as compared to control group).
Kidney functions
The data in table 3 showed no significant changes in urea and serum creatinine in fennel oil group as compared
to control group (Table 3).
Table 3: Changes in BUN and creatinine levels in the control and treated groups
Groups
Parameters
Control
Fennel oil
BUN (mg/dl)
% of change
20.3±0.2
21.6±0.4
6.4%
Creatinine (mg/dl)
% of change
0.7±0.2
0.8±0.01
14.3%
Values represent mean ±SE (stander error).
Body weight and glucose level
Results of the present study showed non-significant change in body weight in the treated group when compared
to control rats and a high significant decrease (P < 0.01) in glucose level in the treated group when compared to
control rats (Table 4).
Table 4: Changes % of body weight and FBS levels in the control and treated groups
Groups
Parameters
Control
Fennel oil
Body weight
% of change
137.6 ± 0.4
148.8±0.4
15.4
FBS (mg/dl)
% of change
94±2.8
86±2.7**
-8.5%
Values represent mean ±SE (stander error). (p**<0.001as compared to control group).
Effect of Phytoestrogen (Fennel) on Some Sex Hormones…
1619
Protein profile
The present study showed that administration of fennel oil to normal rats showed non-significant change in total
protein, albumin, globulin, and albumin/globulin ratio in the treated group when compared to control rats (Table
5).
Table 5: Changes in the total protein, albumin, globulin, albumin/globulin and albumin/globulin levels in the
control and treated groups
Groups
Parameters
Control
fennel oil
Total protein (g/dl)
% of change
6.4±0.11
5.9±0.2
-23.4%
Albumin (g/dl)
% of change
3.7±0.11
3.4±0.1
-10.8%
Globulin (g/dl)
% of change
2.7±0.18
1.5±0.2
-44.4%
Albumin/Globulin(g/dl)
% of change
1.3±0.1
2.2±0.4
% 69.2
Values represent mean ±SE (stander error).
Hormones
FSH and testosterone showed highly significant decrease (p<0.02), while LH showed insignificant change in
fennel oil group when compared to control rats(Table 6).
Table 6: Changes in the FSH, LH and testosterone levels in the control and treated groups
Groups
Parameters
Control
Fennel oil
FSH(ng/ml)
% of change
2.9±0.1
2.2±0.1**
-24.1%
LH(ng/ml)
% of change
1.9±0.1
1.7±0.1
-10.5%
Testosterone(μu/dl)
% of change
3.9±0.2
2.9±0.2**
-17.9%
Values represent mean ±SE (stander error). (p*<0.05, p**<0.001as compared to control group).
Sperm count
The data in table 7 showed decreased in sperm count in fennel oil group when compared to control rats.
Table 7: Sperm count in the control and fennel oil group.
Groups
Parameters
Control
Fennel oil
Sperm count*106/ml
% of change
100±0.01
56±0.01
-44%
DISCUSSION
In the present study fennel oil lead to decrease in
cholesterol, triglycerides and LDL-c levels.
Antioxidative properties and radical scavenging
activity may be the possible mechanisms by which
fennel ameliorated the total lipids, cholesterol,
triglycerides and LDL-c(10).Anethole (t-anethole) that
is the main compound in all fennel volatile oils
possesses significant antioxidant activity. The
presence of t-anethole and flavonoids content in
fennel might be associated with lowering total lipids,
Eman Helal et al.
1620
cholesterol, triglycerides and LDL-c levels. So fennel
is suggested to be a new alternative for clinical
management of hyperlipidemic patients (11).
In the present study fennel oil lead to decrease in
glucose level. In a study by Parsaeyan(12), he showed
marked improvements of hyperglycemia and
pathological changes induced by streptozotocin after
fennel ingestion, which can prove its effect as
antidiabetic in folk medicine. Furthermore, Anitha et
al. (13)suggested that the fennel exhibited the
antidiabetic activity by decreasing oxidative stress and
preserving pancreatic beta-cell integrity(14).The
hypoglycemic effect of fennel might be due to the
presence of triterpenes, steroids, saponins and
phenolic compounds(15).
In the present study, fennel oil lead to decrease
in FSH, testosterone and sperm count.
Spermatogenesis is stimulated by androgen but
inhibited by estrogen and progesterone (16).Anethole is
a phytoestrogen found in fennel and has an estrogen
like effects, it can decrease blood testosterone level
through Sertoli cells that can convert testosteroneto
estradiol and secrete inhibin, which suppresses FSH
synthesis leading to decline in testosterone level. So,
this decline finally causes a decrease in
spermatogenesis (17).
In a study by Zondekand Bergmann (18), they
found that polyanethol compounds that exist in fennel
oil are five times more active than pure anethole.
Anethole possesses anti-spermatogenic effects and
reduces sperm concentrations in epididymis in adult
male albino rats (19). The hormonal inhibition leads to
the decrease in sperm count and sperm motility as a
result of low androgen levels (20).
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