Ameliorative potentials of a combination of fenugreek and α tocopherol on cadmium induced testicular

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Ameliorative Potentials of a Combination of Fenugreek and α-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]. α-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 α-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 α-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 α-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 α-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 α-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 α-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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