Effect of silymarin on DNA and nuclear integrity of ram sperm Treated with sodium arsenite
Volume 7, Issue 4, Spring 2017, Pages 429-436
https://doi.org/10.52547/JCT.7.4.429
i F Eskandar, i HR Momen
Abstract Aim: This study was performed to investigate if silymarin can prevent the adverse effects of sodium arsenite on ram sperm DNA and nuclear integrity.
Material and Methods: Epididymal sperm obtained from Farahani’s ram (Ovis aries) was swim up and divided into five groups: 1. Sperm at 0 hour, 2. sperm at 180 minutes (control), 3. sperm treated with sodium arsenite (10μM) for 180 minutes, 4. sperm treated with silymarin (20 μM) + sodium arsenite (10 μM) for 180 minutes and 5. sperm treated with silymarin (20 μM) for 180 minutes. Ram´s sperm DNA integrity was assessed by SCD (Sperm Chromatine Dispersion) test to study DNA fragmentation and Acridine orange staining was used to estimate DNA denaturation (double-strand DNA versus single-strand DNA). To evaluate sperm nuclear integrity, Diff-quick staining was used and sperm nuclear diameter was measured.
Results: DNA fragmentation percent and nuclear diameter of the spermatozoa were significantly increased and decreased, respectively, in sodium arsenite group compared to the control samples. While this toxicant had no effect on sperm DNA denaturation. In silymarin + sodium arsenite group, silymarin was able to significantly ameliorate the adverse effects of sodium arsenite on these sperm parameters compared to sodium arsenite group.
Conclusion: Silymarin as a potent antioxidant could compensate the adverse effects of sodium arsenite on DNA fragmentation and nuclear diameter of ram sperm.
Apoptosis Will be Increased after 2h Incubation of Human Sperm at 37°C
Volume 4, winter, Winter 2014, Pages 415-423
https://doi.org/10.52547/JCT.4.4.415
Abstract Aim: The main goal was to evaluate the impact of different incubation time intervals on human sperm DNA status using terminal deoxyribonucleotidyl transferase–mediated dUTP nick-end labeling (TUNEL) test.
Material and methods: This prospective study involved 21 normozoospermic specimens. After direct swim-up, sperm cells were incubated at 37°C and DNA damage was evaluated at different time intervals (0, 1, 2 and 3 h). After slide fixation with methanol 100% for 4 minutes and rinse in phosphate-buffered solution (PBS), TUNEL was added to each slide and incubated for 60 minute in dark. Eosin-Nigrosin and Papanicolaou staining protocols were applied in order to assess sperm viability and morphology, respectively.
Results: Sperm viability and normal morphology was improved after sperm processing (100%, 72.3% respectively, p < 0.0001). The rate of DNA damage was significantly higher after 2h compared to 0h (9.19±0.8% Vs 4.9±0.9%, respectively, p=0.008). Also there was significant difference in abnormal sperm DNA between 3h and 1h (10.95±0.7% Vs 7.1±1%, respectively, p=0.020).
Conclusion: Incubation of prepared normozoospermic samples at 37°C more than 2 h may be associated with sperm DNA fragmentation. Therefore, it seems that incubation of human spermatozoa at 37°C should be limited up to 2h prior to use in ART clinics.