الفهرس 
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Abstract
The testes are the primary reproductive organs in male which are responsible for sperm production. They are paired ovoid organs that lie within the scrotum outside the body cavity.
Tramadol is a synthetic centrally acting analgesic. It is an opioidpain medication used to treat mild to severe pain, both acute and chronic e.g pain occurring in diabetic neuropathy and postherpetic neuralgia. It is also used in premature ejaculation and used as antidepressant.
Long term administration of tramadol can affect many vital organs e.g. brain, liver, kidney and testes.The effect on testes may be due to the fact that tramadol increases the testicular levels of nitric oxide and lipid peroxidation and decreased the anti-oxidant enzymes activities significantly.
It is very important to search for protective substance that minimize the toxic effect of tramadol.
Vitamin C is a natural antioxidant acting as free radical scavenger.It stops production of reactive oxygen species so it can be used to prevent toxic effects of tramadol on testes.
The aim of this work is to throw some light on the histological, histochemical and ultrastructural effects of tramadol on the testes of adult male albino rats and to clarify the possible protective effect of vitamin C.
Material & Methods
Sixty (60) adult male albino rats weighing nearly 180-200 gm were usedin the present study.The animals were randomly divided into four main
groups:
group I (control group):
Includes 10 animals & received 0.1ml of physiological saline solution by oral route for 4weeks.
group II (vitamin C treated group):
Includes 10 animals. They received vitamin C at a dose of 500mg /kg/day orally for 4 weeks.
group III (tramadol treated group):
Includes 20 animals and subdivided into two subgroups:
Subgroup III a: Includes 15 animals. They received tramadol at a dose of 50 mg/kg/day (nearly 10 mg/rat) orally for 4 weeks.
Subgroup III b: Includes 5 animals. They received tramadol at a dose of 50 mg/kg/day orally for 4 weeks then left without treatment for another 2 weeks.
group IV (tramadol and vitamin C treated group):
Includes 20 animals. They received tramadol at a dose of 50mg/kg/day& vitamin C at a dose of 500mg/kg/day for 4 weeks.
At the end of determined period of each group, the animals from all groups were anaesthetized using ether inhalation sacrificed, then specimens were taken from the lungs of rats of each group.
All specimens were subjected to the following techniques:
Haematoxylin & eosin stain.
Masson’s trichrome stain.
Periodic Acid Schiff (PAS) stain.
Electron Microscopic technique.
Results & Discussion
The following data have been observed in the present study:
group I (control group):
The control rats showed the well-known histological picture of rat testis. The testis consists of multiple rounded to oval seminiferous tubules. Each tubule is bounded by a basal lamina and lined by spermatogenic and Sertoli cells. Inbetween the tubules, there is a testicular interstitium containingmyoid cells, blood vessels together with clumps of Leydig cells.
PAS reaction was intense in the basal lamina and strong in the interstitium, spermatogenic cells and sperms.
Collagen fibers are normally distributed in tunica albuginea surrounding the testis while minimal amount of collagen fibers is present in the interstitium and around blood vessels byMasson’s trichrome stain.
group II (vitamin C treated group):
Sections of the testicular tissue of this group showed the normal histlogical and histochemical picture the same as group I.
group III (tramadol treated group):
Subgroup IIIa:
Showed irregularity of the seminiferous tubules with disorganization & degeneration of spermatogenic cells. The damaged spermatogenic cells were exfoliated in the lumen which show absence of sperms. Vacuoles appeared between the degenerated spermatogenic cells replacing them. There is also irregular degenerated basement membrane of semiuiferous tubules. Some seminiferous tubules were ruptured. Multinucleated giant cells appeared in the lumen of some tubules. Widening of the interstitium with appearance of homogenous acidophilic exudate and congestion of blood vessels were also noticed.
PAS reaction was mild in degenerated basement membrane and interstitium. Weak reaction was observed in degenerated spermatogenic cells.
There was increase in connective tissue fibers noticed by Masson’s trichrome stain in our study in tramadol treated group in the form of extensive thickening of tunica albuginea, fibrosis of interstitium & fibrosis around the blood vessels.
Subgroup IIIb:
The recovery group showed signs of improvement after cessation of tramadol administration for 4 weeks. The signs of improvement were in the form of increased spermatogenic cell layer with increased sperm concentration in the lumen of the seminiferous tubules. Most of seminiferous tubules were nearly normal but few tubules were still affected after tramadol withdrawal. The affected tubules showed absence of sperms from the lumen with presence of some vacuoles between the spermatogenic cells. Mitotic activities were seen indicating regeneration.
PAS reaction was moderate in the basement membrane and the interstitium. Mild reaction was observed in spermatogenic cells.
There was moderate amount of collagen fibers in the interstitium with mild thickening of tunica albuginea noticed by Masson’s trichrome stain.
group IV (tramadol and vitamin C treated group):
The histological study of the testis of tramadol and vitamin C treated group showed that most of seminiferous tubules appeared normal with normal spermatogenic cell and a lumen full of sperms. Only few tubules were affected with no sperms in the lumen. Some congestion of blood vessels was present.
Moderate to strong positive PAS reaction in the basement membrane of seminiferous tubules, spermatogenic cells, sperms and interstitium was observed.
Some increase in collagen fibers in tunica albuginea, in the interstitium and around the blood vessels were present as noticed by Masson’s trichrome stain.