The protective effects of folic acid and propolis against the toxic effects of valproic acid in prenatal testis tissue

The purpose of this study was to show, using histopathological and biochemical methods, whether changes occurring in the testis tissue of rat pups exposed to the effect of valproic acid (VPA) in the prenatal period can be eliminated using folic acid (FA) and propolis. Eight Sprague Dawley pregnant rats were divided into four groups. No procedure was performed on Group 1 (control group). However, 300 mg/kg/day VPA was administered to Group 2 (VPA group), 300 mg/kg/day VPA+400 µg/day folic acid to Group 3 (VPA+FA group) and 300 mg/kg/day VPA + 100 mg/kg propolis to Group 4 (VPA+propolis group) throughout the experiment. All applications were performed on days 6-21 of pregnancy and at the same time every day. No application was performed on newborn rats after birth, and the study continued with male rat pups. The pups were subsequently divided into four sub-groups of six rats each; Newborn Group 1 (NB-G1), Newborn Group 2 (NB-G2), Newborn Group 3 (NB-G3) and Newborn Group 4 (NB-G4). At the end of the experimental period (postnatal day 21), all animals were sacrificed, and their testes were removed. Testis tissue specimens were evaluated histopathologically under a light microscope. The terminal deoxynucleotidyl transferase (TdT) deoxyuridine triphosphate nick end labeling assay (TUNEL) technique was used to evaluate apoptosis in the testis. Oxidative stress parameters in blood and tissue were examined biochemically. Histopathological examination of VPA group testis sections revealed intense edema in the interstitial area, non-matured germinal epithelial cells in the lumen, occasional vacuolization in germinal cells and a decreased germinal epithelial thickness. Findings such as edema in the interstitial area, non-matured germinal epithelial cells in the lumen and impaired seminiferous tubule structure and separation of the germinal epithelium from the basal membrane in rare areas were observed in VPA+FA group testis tissue. Edema in the interstitial area decreased, but maintained its density. Other findings decreased more markedly. In the VPA+propolis group, findings such as edema in the interstitial area, non-matured germinal epithelial cells in the lumen and separation of the germinal epithelium from the basal membrane were observed. However, these findings decreased compared with the VPA group, the edema in the interstitial area decreasing particularly markedly. Germinal epithelial thickness decreased statistically significantly at morphometric measurement, but increased in the treatment groups, slightly more markedly in the propolis group. The apoptotic index (AI) increased significantly in the VPA group compared to the control group, and although they tended to decrease in the treatment groups, this was not statistically significant. Biochemically, the oxidative stress index (OSI) increased significantly in VPA group tissue biochemistry compared to the control group. Catalase (CAT) values in the VPA+FA group increased significantly compared to the control group. Tissue malondialdehyde (MDA) decreased significantly compared to both the control and VPA groups. Glutathione (GSH) decreased significantly compared to the control group, while total oxidant status (TOS) and OSI increased. Serum total antioxidant status increased significantly compared to the control and VPA groups. In the VPA+propolis group, TOS and OSI increased significantly compared to the control group. Our findings suggest that VPA applied in the prenatal period adversely affects rat pup testis development, while the addition of folic acid and propolis can produce an improvement in histopathological and biochemical findings.