Alleviation of lead acetate-induced testicular dysfunction and related toxicity changes in rats using Nigella sativa L. seeds

Lead acetate (LA) toxicity can occur either through ingestion or inhalation from contaminated surfaces or from the environment. N. S ativa is a natural product with immense pharmacological properties, which include antioxidant, antibacterial, anti-anemia and reproductive enhancement properties. Several studies have reported that LA can alter hematological, biochemical, reproductive and oxidative stress in short-term. In this study, the prophylactic and therapeutic effects of N. Sativaon chronic and sub-chronic lead acetate induced hematological-biochemical changes, histopathology and influence on male reproductive hormonal levels were evaluated. In the first experiment (therapeutic study), a total of 75 male Sprague-Dawley rats were divided into three groups with 25 rats in each group and in this single group, it was further sub-divided into five groups of 5 rats each. Group 1 acted as the negative control and were given distilled water, group 2 acted as the positive control (PC) and were given 10 mg/kg of lead acetate (LA) orally/daily, group 3 (T1), 4 (T2) and 5 (T3) were each given LA 10 mg/kg and graded concentrations (100 mg/kg, 150 mg/kg and 200 mg/kg) of N. S ativa orally, respectively. Twenty-five rats were euthanized at day 30, 60 and 90, respectively, for the collection of whole blood, serum, and organs. In the second experiment (prophylactic study), a total of 20 male Sprague-Dawley rats were divided into four groups of five mice each. Group 1 (NC) was the negative control, group 2 was the positive control (PC) and was administered 10 mg/kg/per day of lead acetate (LA) per OS, group 3 (T1) was administered 200 mg/kg/daily of N. S ativa per OS for one month and Group 4 (T2) was pre-treated with 200 mg/kg/daily of N. S ativa per OS for one month, followed by administration of 10 mg/kg/daily of lead acetate (LA) alone per OS for another month. At the end of the experiment, whole blood, serum and tissue were collected to evaluate the complete blood profile, serum biochemistry, hormonal concentration, spermiogram and histopathological changes. For the determination of the spermiogram, the right epididymal segment was collected and homogenized in phosphate-buffered saline, and the homogenates were used for the aforementioned purpose. The haemogram from the therapeutic study showed a lower (p<0.05) red blood cell count, packed cell volume (PCV) and mean corpuscular hemoglobin concentration (MCHC) in the PC and T1 groups, while T2 and T3 have normal haemograms. Biochemical analysis revealed an elevated (p<0.05) aspartate aminotransferase (AST), alanine aminotransferase (ALT) and creatinine levels in the PC and T1 groups on day 90 for AST and day 30 for ALT and creatinine respectively. The level of alkaline phosphatase (ALP) was higher (p<0.05) in the PC at 30 and 60 days of sampling. Other parameters such as WBCs, prothrombin, urea and cholesterol were not significantly altered in all groups. Histopathological lesions in the liver and kidneys were more severe in the PC and T1 groups, while the T2 and T3 groups showed mild lesions. There was a decrease (p<0.05) in the total PAS stained area signifying glycogen depletion in the PC, T1 and T2 groups at 60 days, and a higher distribution of PAS stained areas (p<0.05) in the T3 group. At 90 days, the PC group had a lower (p<0.05) distribution of PAS stained areas in comparison to the other groups. There was reduced spermatogenesis and epididymal sperm reserves in the PC group in comparison to the treatment groups. The level of testosterone concentration was lower (p<0.05) in the PC group at 90 days, while FSH was lower (p<0.05) in T3 at 30 days and LH was higher (p<0.05) in T1 at 90 days. Estradiol concentration was higher (p<0.05) and comparable between the control and T3 at all sampling points. There was a decreased level of superoxide dismutase (SOD) and total glutathione (GSH) in the PC group, and an increased GSH level in the T3 group at all sampling points. The spermiogram showed an increase in sperm concentration, viability, and motility in the treated group as compared to the PC. Furthermore, the PC had a higher (p<0.05) incidence of sperm abnormalities. The haemogram from the prophylactic study showed lower (p<0.05) level of hemoglobin, PCV, and prothrombin in the PC, while WBC, band neutrophil, segmented neutrophil, lymphocyte and monocyte counts were higher (p<0.05) in the PC than the treatment groups. However, eosinophil count was higher in T2, while no changes were observed in RBC and MCV values. Both AST and ALT were higher on the PC when compared to other groups. Similarly, the levels of ALP, cholesterol, urea, and creatinine were all higher (p<0.05) in the PC group and comparable (p>0.05) in the control, T1, and T2 groups. The sperm concentration, general and individual motilities were higher (p<0.05) in the NC and T1 animals, while the T2 had intermediate and the PC had lower (p<0.05) values of each parameter. Percentage of sperm viability was higher (p<0.05) in the T1 and lower (p<0.05) in the PC group. However, percentage abnormality was lower in T1, comparable in NC and T2, and higher (p<0.05) in PC. Spermatogenic cell population and epididymal sperm reserve (ESR) were optimal in control and pre-treated animals, while the PC had lower spermatids and ESR. The concentration of estradiol was lower (p<0.05) in the PC and T2, while LH concentration was lower (p<0.05) in the PC, and comparable (p>0.05) between control and T2. The concentration of FSH was comparable (p>0.05) in all groups, while TSH concentration was lower (p<0.05) in the PC and higher in the control and T1 groups. The level of SOD and GSH were lower (p<0.05) in the PC and T2 groups. In summary, this study has shown the prophylactic and therapeutic potentials of N. Sativa seed extract in both sub-chronic and chronic lead acetate induced toxicity in the male reproductive system in rats.

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