This study demonstrated that Dex caused marked histological changes in testicular structure. This might lead to a disturbance in the process of spermatogenesis. The surrounding basal lamina of the seminiferous tubule was thickened, irregular, and wavy. Similar changes were found in different types of testicular damage and also after hypophysectomy 21,22. It was suggested that this basal lamina thickening is the result of either the overproduction of collagen fibers by fibroblasts or the decrease in the rate of collagen removal 23. It was found that Dex increased the amount of collagen fibers, especially type IV, through its effect as a metalloproteinase inhibitor that suppresses extracellular matrix degradation 24. It was also proved that Dex promotes connective tissue growth factor production and activation, which stimulates fiber formation by fibroblasts 25,26. Furthermore, some studies found that Dex induced overproduction of reactive oxygen species by different cells such as macrophages and vascular endothelium 27,28. These species could increase the production of collagen fibers and induce fibrosis 29. It is well known that the basal lamina plays an important role in maintaining substance transmission between the interstitial tissue and spermatogenic epithelium and in maintaining the structural and functional integrity of these tissues 30.
In the present work Dex induced degenerative changes in most of the germ cells in the form of increased chromatin texture and cytoplasmic density, ballooned mitochondria with destructed cristae, and dilated SER cisternae. The spermatogonia were the most affected cells; this might have been due to their proliferating character, which makes them the target for the toxic effect of Dex. These results are in agreement with other studies that reported an increase in many apoptotic markers such as Bax and Fas ligand expression and TUNEL reactivity in spermatogonia 31. These degenerative changes in germ cells might result from increased oxidative stress production of malondialdehyde in testicular tissue that was detected by other authors in plasma and aorta after Dex administration 32. There were also structural abnormalities in round and late spermatids in the present work. Some round spermatids exhibited irregular nuclei, abnormal acrosome, and numerous mitochondria. Other late spermatids had abnormally angled heads and abnormal Y tails. However, these changes might be secondary to Sertoli cell injury that affected the synthesis of certain proteins essential for germ cell differentiation. It was found that these proteins are secreted at their highest rate during spermatid elongation and spermiation 33. This could explain the highly significant increase in the percentage of sperm abnormalities and their ultrastructural defects. Furthermore, it was found that degenerative changes in germ cells and the release of reactive oxygen radicals produced varieties of morphologically abnormal sperms and caused DNA damage 34,35. In addition, the presence of numerous mitochondria might be attributed to the high-energy consumption of the detoxification process 36. These results are in agreement with those of previous researchers who found low concentrations of free radical scavenging enzymes in the sperm cytoplasm and hence it was more vulnerable to Dex toxicity 37.
Ultrastructural features of Sertoli cells revealed interrupted junctions, marked nuclear enfolding, vacuoles, irregular secondary lysosomes, and numerous lipid droplets. The desquamated germ cells and the intercellular spacing detected in this work might be secondary to Sertoli cell injury and subsequent retraction of their cytoplasmic processes. This led to disruption of cell arrangement, which became easily sloughed out. Similar findings were reported by other authors who explained the luminal shedding of germinal cells as alteration of intercellular junctions between Sertoli cells 38. Moreover, it was suggested that Sertoli cells secrete Glial cell line-derived neurotrophic factor, which is known to influence the fate of undifferentiated spermatogonia. They reported that Sertoli cell injury was associated with decreased production of this factor, which led to apical sloughing of desquamated spermatocytes or spermatids into the lumen and a massive loss of germ cells 39,40.
Multinucleated giant cells were observed in some of the seminiferous tubules and comprised mainly round spermatids in the Dex-treated group. Similar findings were also seen in the testis of prepubertal normal rats, in old people, and under toxic conditions 41. It was found that these cells formed as a result of the breakdown in the intercellular bridges that connect groups of each specific cell type of the germinal lineage 42. However, it was also reported that such multinucleated cells could be considered a sign of apoptosis 43.
The present results revealed inflammatory signs in the interstitial tissue. There were acidophilic exudates, apparent hyperplasia of interstitial cells, and congested blood vessels. It was found previously that at the site of tissue injury activated innate immune cells release various chemical mediators that are responsible for inflammation 44. Furthermore, the molecules derived from plasma proteins and cells in response to tissue damage could mediate inflammation by stimulating vascular changes and leukocyte migration 45.
Degenerative changes in some Leydig cells were detected in Dex-treated animals in the form of irregular small-sized nuclei with marginated heterochromatin, dilated cisternae of the SER, large lipid droplets, and vacuoles. It was suggested that the degenerative changes in the germ cells may occur secondary to their deprivation of testosterone. This could be a result of excess production of reactive oxygen radicals, which lead to disruption of the mitochondria and inhibition of steroidogenic acute regulatory protein expression 46. In addition, it was stated that corticosteroids decrease the Leydig cell sensitivity to gonadotropins, either by reducing the LH receptor content or by inhibiting the 17α-hydroxylase and/or C17, 20-lyase activity 47–50.
There are no conflicts of interest.
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