Pumpkin seed and grape seed extracts ameliorate cyclophosphamide-induced spermatogenesis inhibition in rat model: Histomorphometrical, immunohistochemical, and ultrastructural approaches

Abstract

This study investigated the protective efficacy of grape seed extract (GSE) and pumpkin seed extract (PSE) against cyclophosphamide (CP)-induced testicular toxicity in adult male albino rats, focusing on biochemical, histopathological, histomorphometrical, ultrastructural, and immunohistochemical perspectives. Thirty-six male albino rats were divided into six groups: control, PSE, GSE, CP, CP + PSE, CP + GSE. Testicular tissues and serum samples were collected for analysis after 8 weeks. The CP group exhibited significant testicular damage with distorted seminiferous tubules (marked decline in diameters and germinal epithelial thicknesses), germ cell apoptosis, and disrupted spermatogenesis (decline in Johnsen’s score and weak immunoexpression of PCNA in basal germ cells). Ultrastructurally, germ cells exhibited shrunken pyknotic nuclei and cytoplasmic aggregations of ribosomes with dilated or shrunken mitochondria. Spermatozoa displayed an irregular axoneme with loss of fibrous sheath. Furthermore, there were significantly decreased testosterone levels and increased oxidative stress markers (elevated malondialdehyde levels and decreased glutathione levels).  In contrast, the CP + PSE and CP + GSE groups showed marked mitigation of these changes, resulting in a notable amelioration of oxidative stress markers, and testosterone levels, preservation of testicular architecture, and normalization of spermatogenic processes, along with elevated Johnsen score. Immunohistochemical analysis revealed an increase in PCNA in the CP + PSE and CP + GSE groups compared to the CP group. These findings suggested that GSE and PSE, owing to their antioxidant properties, could effectively reduce CP-induced testicular damage.