Research Journal of Medicine and Medical Sciences, 4(2): 386-390, 2009

© 2009, INSInet Publication

Effects of Black Seeds (Nigella Sativa) on Spermatogenesis and

Fertility of Male Albino Rats
1
Mukhallad A Mohammad, 1 Mohamad MJ Mohamad and 2 Hatham Dradka.

1
Physiology Dept., Faculty of Medicine, Jordan University of Science and Technology, P.O.
Box 3030, Irbid 22110, Jordan
2
Department of Biology, Faculty of Science, Jerash Private University, P.O. Box 311, Jerash
26110, Jordan.

Abstract: The seeds of Nigella sativa Linn. (Ranunculaceae herbaceous plant), commonly known as black
seed or black cumin, are used as herbal medicine all over the world for the treatment and prevention of
a number of diseases and conditions like asthma, diarrhoea and dyslipidaemia. The aim of this study is
to see effects of Nigella sativa on the reproductive system and fertility on adult male albino rats. 20 albino
rats were involved in this study and were divided into two groups: a vehicle-treated control group and
a treated group with Nigella sativa at a dose of 300.mg/kg body weight for 60 days. The seeds of
Nigella sativa induce a significant increase in the weight of reproductive organs as compared to control
animals (P<0.01). The sperm motility and count in cauda epidydimides and testicular ducts were
significantly increased (P<0.01). Spermatogenesis was increased at primary &secondary spermatocyte
stages. Epididymides showed eleveted number of spermatozoa. Lumen of vas deferentia were full of
sperms. The secretary activities of seminal vesicle and ventricular prostate were also increased. A
significant increase (P<0.01) in spermatogenesis activity was observed in semniferous tubule. Treated rats
testicular cell population showed a increase in number of spermatocytes and spermatids (P<0.001) when
compared to control animals. Increased in number female rats impregnated by males receiving treatment
was also observed and (P<0.01). It is concluded that the aqueous extracts of Nigella sativa have increased
spermatogenesis of male albino rats.

Key words: Nigella sativa, Fertility, spermatogenesis, male and female albino rats.

INTRODUCTION contained within are exposed to the air, becoming

The use of plants as medicines dates from the
earliest years of man’s evolution [1 ] . Medicinal plants
serve as therapeutic alternatives, safer choices, or in
some cases, as the only effective treatment. People in
different cultures and places have used particular plants
for to treat certain medical problems. A larger number
of these plants and their extract have shown beneficial
therapeutic effects, including anti-oxidant, anti-
inflam m ato ry, anti-cancer, an ti-m ic ro b ia l, a nd
immunomodulatory effects[1-6 ] . Among the promising
medicinal plants, Nigella sativa, a dicotyledonous of
the Ranunculaceae family, is an amazing herb with a
rich historical and religious background [7 ]. Nigella
sativa is found in southern Europe, northern Africa, and
Asia Minor. It is a bushy, self-branching plant with
white or pale to dark blue flowers. Nigella sativa
reproduces with itself and forms a fruit capsule which
consists of many white trigonal seeds. Once the fruit
capsule has matured, it opens up and the seeds
black in color[8 ] . The seeds of Nigella sativa are the
source of the active ingredients of this plant. It is the
black seed referred to by the prophet Mohammed as
having healing powers[7 ] . Black seed is also identified
as the curative black cumin in the Holy Bible and is
described as the Melanthion of Hippocrates and
Discroides and as the Gith of Pliny[9 ].
Historically, it has been recorded that Nigella
sativa seeds were prescribed by ancient Egyptian and
Greek physician to treat headache, nasal congestion,
toothache and intestinal worms, diuretic and to increase
milk production[7 ,1 0 ] . The seeds of Nigella sativa, known
as black seed, black cumin have long been used in the
middle and far east as a traditional medicine for a wide
range of illnesses including bronchial asthma, headache,
dysentery, infections, obesity, back pain, hypertension
and gastrointestinal problems[8 ,1 1 ] . Its use in skin
condition as eczema has also been recognized
worldwide [7 ] .
Four dolabellane-type diterpene alkaloids have been

Corresponding Author: Mohamad MJ Mohamad MD, Physiology Dept., Faculty of Medicine, Jordan University of
Science and Technology, P.O. Box 3030, Irbid 22110, Jordan
E-mail: mukmoh@just.edu.jo Mobile: (00962) (079) (5158960)
386
 Res. J. Medicine & Med. Sci., 4(2): 386-390, 2009
isolated from the seeds of N. sativa [1 2 ,1 3 ] . High
performance chromatographyic analysis of Nigella
sativa oil showed many compounds like thymoquinone,
dithymquinone, thymohydroquinone and thymol[1 4 ]. N.
sativa seeds contain other ingredients, including
nutritional components such as carbohydrates, fats,
vitamins, mineral elements, and proteins, including
eight of the nine essential amino acids [1 4 – 1 8 ] .
Monosaccharides in the form of glucose, rhamnose,
xylose, and arabinose are also found. Nigella sativa
seeds are rich in the unsaturated (linoleic and oleic
acid ) and essential fatty acids.[1 4 ,1 5 ] . The major
p h o sp h o lip id cla sse s a re p ho sp hatid ylcholine,
phosphatidylethanolamine, phosphatidyl-serine, and
p h o sp h atitd ylinisito l [ 1 4 ,1 5 , 1 8 ] . T he seed s contain
carotene[15 ] calcium, iron, and potassium [1 9 ] . In addition
to the previous nutritional values, the Nigella sativa oil
has a wide margin of safety[20 ] In the light of the above
findings this work was conducted to monitor its effect
on reproductive system and fertility in adult male rat.
M ATERIALS AND M ETHODS
Animals and treatment: Adult male and female
albino rats of Sprague Dawley strain, weighing about
250-300 gm. were raised in the Animal House Unit in
Faculty of Agriculture and Science at Jerash Private
University under controlled temperature of 21 ±1C and
12 hours light, 12 hours darkness schedule (lights on
06.00 AM–18.00PM). Food and W ater were avialable
ad labitum.
Plant material: Seeds of Nigella sativa were
purchased from the local market in irbed. The seeds
was grinded with a grinder into powder and dissolved
in normal saline (freshly prepared) to a final
concentration of 200 mg/ml. The Aqueous extracts of
Nigella sativa is administered orally to rats (300
mg/kg) using animal feeding intubations needles.
Experimental Design: Male rats were divided into
following groups.
Group 1- (20 rats) a control group, the rats in this
group received vehicle (Normal Saline) for 60 days.
Group 2 – (20 rats), the rats in this group received
orally an aqueous extract of Nigella sativa at dose of
300-mg/kg body weight for 60 days.
Fertility was estimated in adult male rats treated
with aqueous extracts of Nigella sativa and in the
control males counterparts. Each male was placed in an
individual cage with two virgin untreated females of
the same strain. They were left together for 10 days
during which two estrous cycles had elapsed (21). One
week after the removal of the exposed males, pregnant
females rats were killed by cervical dislocation under
387
light ether anesthesia and the number of implantation
sites, the number of viable fetuses and the number of
resorption sites were recorded. After 24 hours of the
last dose, male rats were weighed and killed by
cervical dislocation under light ether anesthesia. The
reproductive tract was taken out trimmed free of fat
and each organ was weighed separately on electronic
balance. The reproductive organ taken into account for
study in male include testes, epididymides, ventral
prostrate, seminal vesicle, and vas deferens. Small
pieces of the above tissues the obtained were fixed in
Bouin’s fixative for histological studies. After
dehydration using different concentration of alcohol,
specimens were embedded in paraffin blocks and
sectioned at 5µm, placed on a clean histological glass
slide and stained using Haematoxyline and Eosin stain
Sperm Motility and Count: To determine the sperm
count and motility, a 100mg of cauda epididymides
was minced in 2 ml of physiological saline and one
drop of the evenly mixed sample was applied to a
Neubauer's counting chamber under cover slip.
Quantitative motility expressed as percentage was
determined by counting both motile and immotile
spermatozoa per unit area. Cauda epididymal and
testicular sperm counts were made by routine procedure
and expressed as million/ml of suspension (22).
Histometry: W ith the help of Camera Lucida, one
hundred of circular appearing somniferous tubules were
traced at x80 magnification and the diameter of each
tubule was measured separately. The measurement was
expressed in terms of mean of all the traced tubules.
Similarly, Leydig cell and their nuclei were traced at
x800. In addition, epithelial cell height of cauda
epididymides, caput epididymides and seminal vesicle
were also traced at X-360 and recorded.
Testicular Cell Population Counting: Spermatogenic
elements namely spermatogonia, spermatocytes and
spermatids were counted in 5 um thick cross sections
of 10 semniferous tubules obtained from 10 animals of
each group. Interstitial cell types (such as fibroblast,
immature and mature Leydig cells and degenerating
cells) were estimated, applying a differential count over
200 cells population and statistically verified by the
binomial distribution (23).
Statistical Calculation: All the values of body/organ
weight biochemical estimation and histometry were
expressed in terms of mean value ± S.D. The different
treatment groups were compared with control group
using chi-square test and Student’s “t” test
RESULTS AND DISCUSSION
Table.1 shows no signifcant increase in the body
weight after adminstration of aqueous extracts of
 Res. J. Medicine & Med. Sci., 4(2): 386-390, 2009

Nigella sativa while the weight of the testes,
epididymides, seminal vesicle, ventral prostate and vas
deferens were significantly increased(P< 0.01) in
treated male rats compared to control group.
Table. 2 shows that the motility of sperm in cauda
epididymis was significantly (P< 0.01) increased in
treated animals that Nigella sativa in comparison with
control. Sperm density in treated animals, the
seminiferous tubule diameter and leydig cell nuclear
diameter of treated male was increased significantly
(P< 0.01). Epithelial cell height in epididymides (cauda
and caput) and seminal vesicle were significant
increased (P< 0.01).
Table.3 shows that the administration of Nigella
sativa extract caused a significant increased in the
germinal cell population: spermatocytes (primary and
secondary) and spetmatids were decreased to significant
level (P< 0.001). Similarly the immature and mature
L e y d i g c e lls n u m b e r w e r e a l s o i n c r e a s e d
significantly.However the degenerating cells number
was significantly decreased (P< 0.001). increased
fibroblast and spermatogonia numbers were not altered
significantly.
(Table.4) The results presented in table shows that
oral administration of (Nigella sativa) at dose (300
mg/kg body weight) for 60 days to male rats had
significant increased (P< 0.01) on the number of
females impregnated by them. The number of
implantations and number of viable fetuses were
significantly (P< 0.01) increased in female rats
impregnated by males ingested Nigella sativa. The
number of implantations (P< 0.01)and number of viable
fetuses (P< 0.01) were also significantly increased in
females impregnated by males ingested Nigella
sativa.also the number of resorptions was significantly
(P< 0.05) decreased in females impregnated by males
ingested (300 mg/kg body weight).
Discussion: The Nigella sativa is currently used by
Jordanian population as a phrodisiac and fertility
promoting agent. The animal model used in this work
has been used previously by several workers to assess
the adverse effects of extract obtainned from medicinal
plants on reproductive functions in male [2 4 ] .
In rats the whole spermatogenic process requires 53
days out which spermatozoa spends last 6 to 7 days in
the final transit through epididymides [2 5 ]. The Nigella
sa tiva was a d m inistra te d fo r o n e c o m p le te
spermatogenic cycle Present investigation shows that
oral administraion of Nigella sativa increased fertility
in male albino rats. The weight of reproductive organs
were markedly increased(Table.1) . The weight,size and
secretory function of testes, epididymes, seminal
vesicles,ventral prostate,vasa deferentia are closly
regulated by androgens [2 6 ,2 7 ] . The drug may act on
pituitary gland and increased main hormone of
spermatogenesis. It is well established fact that weights,
size histological appearance and secretary functions of
the epididymides, seminal vesicle and ventral prostate
are closely regulated by the androgens, changes taking
place in these organs after castration can be
counteracted by administration of testicular hormones
thus serving as "indicator test" for the male hormones
[26 ,2 7 ]
.
The process of spermatogenesis and accessory
reproductive organs function are androgen dependent.
In present study the number of mature Leydig cells
were significantly increased. This reflects the increase
of androgen level. It is further confirmed by increased
number of spermatocytes ( primary and secondary )
and spermatids as these stages are completely androgen
dependent[2 8 ] . The increase weight and histometry of
reproductive organs further confirmed androgen
increase. Significant increase in the sperm motility of
cauda epididymis was observed in treated group. This
may be due to activity effects of Nigella sativa on the
enzymes of oxidative phosphorylation (29).

Table 1: Effects of N igella Sativa (300m g/kg ) on Body and organ weights m ale rats .
Treatm ent Body weight (gm ) Testes Epididym ides Sem inal vesicle Ventral Prostate Vas deferens
----------------------- -------------------------------------(m g/100 gm body weight)-------------------------------------------------------
Initial Final
Control 267± 12.21 296± 11.76 875±8.66 317±3.81 371±9.37 179±5.38 62± 0.66
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
N igella Sativa 275±8.35 304±7.66 925*± 9.83 395*± 8.62 404.58*±5.5 215*±3.01 87*± 1.78
Results are expressed as m ean S.D .
20 rats were included per group.
*p < 0.01, significantly different from control group (Student's "t" test).

Table 2: Effects of N igella Sativa (300m g/kg ) on H istom eterical param eters and sperm dynam ics in m ale rats
Treatm ent Sperm m otility % Sperm count m illion/m l Sem iniferous tubule Leydig cell nuclear Epithelial cell height
------------------------------ -------------------------------------------------
Cauda Testes Cauda diam eter diam eter Caput Cauda Sem inal vesicle
µm
Control 49.23 *** 4.12*** 54.45*** 265.3*** 6.07 *** 32.47 *** 21.23 *** 17.96 ***
±.0.33 ±0.65 ± 1.66 ± 2.8 ± 0.9 ± 1.69 ± 1.77 ±1.08
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
N igella Sativa 94.1** 8.75** 76.0** 314.6** 8.45** 36.8** 36.08** 0.3228.32**
± 1.94 ± 0.47 ± 1.94 ± 3.2 ± 0.96 ± 0.4 ± 0.17
Results are expressed as m ean S.D .
20 rats were included per group.
**p < 0.01 ***p < 0.001 significantly different from control group (Student's "t" test).

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 Res. J. Medicine & Med. Sci., 4(2): 386-390, 2009

Table 3: Effect of N igella Sativa (300m g/kg ) on Testicular cell population dynam ics in m ale rats.
Treatm ent Germ inal cell types Interstitial cell type
--------------------------------------------------------------------------- --------------------------------------------------------------------------------
Sperm atogonia Sperm atocyte Sperm atocyte Sperm atids Fibroblast Im m ature M ature D egenerating cell
(prim ary) (secondary) Leydig cell Leydig cell
control 13.54±1.69 9.25±1.66 12.35±1.96 42.36±1.4 34.32±1.8 36.75±2.11 38.18±1.66 53.16±1.49
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
N igella Sativa 23.99** 18.85** 64.126** 147.71*** 63.83** 65.195** 70.64** 18.34***
± 0.93 ± 0.80 ± 3.51 ± 4.87 ±1.64 ± 3.47 ± 1.03 ± 1.67
Results are expressed as m ean S.D .
20 rats were included per group.
**p < 0.01 significantly different from control group (Student's "t" test).

Table 4: Effect of N igella Sativa (300m g/kg ) on rats fertility in m ale rats.
Treatm ent N o.of m ale N o.of fem ale N o.of pregnant fem ales N o.of im plantation sites N o.of viable fetuses N o . o f r e s o r p t i o n / t o t a l
N o. of im plantation sites
Control group 10 20 17/20 (85% ) 8.73± 2.66 8.77± 1.16 6/148(4.05% )
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
N igella Sativa 10 20 19/20† (95% ) 9.86**± 2.83 9.85 **± 1.67 4/187† (2.13% )
Results are expressed as m ean S.D .
**p < 0.01 significantly different from control group (Student's "t" test). † p < 0.05 (chi -square test)

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