Piperine mitigates oxidative stress, inflammation, and apoptosis in the testicular damage induced by cyclophosphamide in mice.

Although cyclophosphamide (CP) has been approved as an anticancer drug, its toxic effect on most organs, especially the testis, has been established. Piperine (PIP) is an alkaloid that has antioxidant, antiapoptotic, and anti-inflammatory activities. This study was investigated the protective effects of PIP on CP-induced testicular toxicity in the mice. In this experimental study, 48 adult male BALB/c mice (30-35 g) were divided into six groups (n = 8), receiving normal saline (C), 5 mg/kg of PIP (PIP5), 10 mg/kg of PIP (PIP10), 200 mg/kg of CP, 200 mg/kg of CP + PIP5, and 200 mg/kg of CP + PIP10. On the eighth day of the study, blood and testis samples were prepared for serum testosterone hormone quantification, sperm analysis, histological, and immunohistochemical assays. The results of this study showed that CP induced testicular toxicity with the decrease of sperm count, motility, and viability. Also, CP treatment caused histological structure alterations in the testis, including exfoliation, degeneration, vacuolation of spermatogenic cells, and reducing the thickness of the epithelium and the diameter of the seminiferous tubule. In addition, CP decreased glutathione (GSH) levels, increased malondialdehyde (MDA) levels, Caspase-3, and NF-κB. At the same time, PIP treatment reduced testicular histopathological abnormalities, oxidative stress, and apoptosis that were induced by CP. These results showed that PIP improved CP-induced testicular toxicity in mice, which can be related to its antioxidant, antiapoptotic, and anti-inflammatory activities.

Comparative evaluation of the effect of three types of fixatives (formalin, Bouin and Carnoy) on histomorphological features of various viscera.

Tissue shrinkage is one of the problems in preparing tissue sections. This study compares the use of 10% formalin, Bouin and Carnoy as fixatives on several mouse tissues to determine histomorphological features. In this experimental study, liver, kidney, heart, lung, testicle, spleen, brain and cartilage tissues were isolated from five BALB/c mice. Then, they were fixed with three types of fixatives. After dehydrating, clarifying and embedding, all samples were stained with haematoxylin and eosin. Then, the tissue structure of the viscera was evaluated qualitatively. The results showed that each fixative is more suitable for evaluating a specific part of the tissue. However, relative shrinkage appeared in the tissue sections fixed with 10% Formalin, (1) in the heart as spaces between muscle fibre bundles, (2) in the liver as the dilation of the liver sinusoidal spaces, (3) in the kidney tissue as the expansion of the lumens of the convoluted proximal and distal tubules, (4) in the spleen as open spaces inside the red and white pulps and (5) in the brain as an increase in the space between the cells of the granular and pyramidal cell layers of the cortex. In tissues that were soft and fragile, such as testis, liver and brain, Bouin's fixative was more suitable. Carnoy's fixative was more suitable for the spleen and kidney tissue. Based on the study results, formalin and Bouin were more suitable for heart and cartilage tissue. Considering that in the histopathological evaluation both the cytoplasm and the nucleus are evaluated, it is suggested to choose the fixative suitable for the type of tissue.