Ameliorative Potentials of a Combination of Fenugreek and

-tocopherol on Cadmium induced Testicular toxicity: An

Ultrastructural Study
Adel M. Hussein, Hesham N. Mustafa, Mohamed H. Badawoud

Anatomy Department, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia

(EP) pieces can be observed. Fig. (3C). A Leydig

cell (L) with an oval, indented nucleus. An electron dense bodies, vacuoles (V), an empty space
(black star), lymph vessels (Ly), and the ruptured
vacuolated mitochondria can be observed (white
arrow). Spermatogonia (Sg) and Sertoli cell (St)
are resting on an apparently thickened basal lamina (BL) with a myoid cell (star). Fig. (3D). Spermatids (Sd) with rounded or oval nuclei that have
disturbed chromatin and peripherally arranged
the mitochondria with lost cristae (black arrow)
and the characteristic acrosomal cap (Ac) can be
observed.

Introduction

Cadmium is an ecological pollutant which classified amongst the 10 most poisonous elements
for human beings. Due to its long biological
half-life, exposure to cadmium causes toxic
effects after long periods of a cessation of
exposure due to its residual effects. Peroxidation induced by Cd is the cause of the
release of free oxygen radicals. The oxygen
radicals lead to the destruction of sensitive
molecules and tissues [1]. -tocopherol causes
the elimination of the free oxygen radicals and
the inhibition of the peroxidation [2]. Fenugreek seed demonstrate an antioxidant impact
due to presence of different ingredients like
flavonoids, alkaloids and amino acids [3].

Objectives

The purpose of this study is to evaluate the

cytoprotective effcts of fenugreek and -tocopherol on cadmium chloride induced testicular
damage.

Materials & Methods

Forty male adult Wistar rats weighed 175-200

gm, randomly placed into four groups (N=10).
Group I served as the control group. Group II
received 5 mg/kg/day cadmium chloride (1/15
LD50) [4]. Group III received 5 mg/kg/day cadmium chloride and 150 mg/kg/day fenugreek
and 100 mg/kg/day of -tocopherol [5]. The
prophylactic combination started one week
before CdCl2 administration to acquire the
benefis of both curative and preventive regimens. All groups were treated by oral gavage
for 60 consecutive days. Testis where subjected to histology using H&E. Immunohistochemistry (IHC) using Anti-CD68+ for macrophages,
Anti CD3+ for T-lymphocytes, and Anti CD20+
for B-lymphocytes. Morphometry and Ultrastructural examination.

Fig. (2A). Control. Fig. (2B) CdCl2 showed a noticeable reduction in epithelial cells. Vacuolations
(V) and intercellular areas (sp) among the germ
cells can be observed. Fig. (2C) CdCl2 showed
depletion, and separation of the spermatogonia
layer, vacuolization (V) of the germinal epithelium. (2D) Combined fenugreek and -tocopherol
with CdCl2 shows an apparent improvement of
the testicular architecture with a partial restoration of a low germinal epithelium.

Graph (2): Mean Diameter of Seminiferous

Tubules and Germinal Epithelial Height.

Results

Graph (3): Mean area percentage of CD3, CD20,

and CD68 immunostaining.

Graph (1) Mean testes weight and body weight by

grams

Fig. (4A). Combined fenugreek and -tocopherol

and CdCl2 showed a Sertoli cell (St) with a euchromatic-indented nucleus and apparent nucleolus (N) and the spermatogonia (Sg) resting
on the thickened basal lamina (BL) with an apparent myoid cell (black star). Some mitochondria are affected, and others are healthy (arrow).
Fig. (4B). Reconstruction of the structure of the
cross-sections of the mid pieces (MP) with axoneme (white arrow) surrounded by a mitochondrial sheath (black arrow). Fig.(4C). Leydig cells
with oval euchromatic nuclei (L) and peripherally
located heterochromatin. The cytoplasm contains
multiple lysosomes (S) and many mitochondria
(arrow). Fig. (4D). Improvement of the appearance
of germ cells. The Spermatids (Sd) with rounded
or oval euchromatic nuclei and peripherally arranged mitochondria with preserved cristae (arrow) and the characteristic acrosomal cap (Ac)
can be observed. Minimal empty spaces (star) in
the affected intercellular bridge.

Conclusions

Fenugreek and -tocopherol could represent

a promising medicinal combination to ameliorate the toxic effects of cadmium exposure.

References

Fig. (1A): Control. Fig. (1B): CdCl2 showed disintegration of the testicular architecture, vacuolation (V), a separation of the spermatogonia from
the basal lamina (), and a widening of the interstitial spaces (W). Fig. (1C): Higher magnification
showed multinucleated giant cells (G) and congestion of the capillaries (C). Fig. (1D): CdCl2 and
combined fenugreek and -tocopherol showed
preservation of the architecture of the testicular
tissues.

Fig. (3A). CdCl2 group showed a Sertoli cell with

an irregular euchromatic nucleus (St) with the
characteristic indentation of the nucleus. Primary spermatocyte (Sc) is separated by intercellular spaces (white star) and the vacuolated distorted mitochondria (black arrows). Vacuoles (V) all
over the cytoplasm. Note the apparently thickened basal lamina (BL) containing collage (black
star) and a myoid cell (curved arrow). Fig. (3B).
A mid piece of the sperm (MP) with distorted and
swollen mitochondrial sheaths (black arrow) surround the central axoneme (white arrow). End

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