Dose- and Time-Dependent Effects of Quercetin on Apoptosis and Oxidative Stress in Post-Thaw Ram Spermatozoa
| Author | Eshagh Abangah | en |
| Author | Mohammad Zandi | en |
| Author | Mehdi Aminafshar | en |
| Author | Annahita Ghaedrahmati | en |
| Orcid | Mohammad Zandi [0000-0002-0523-1719] | en |
| Orcid | Annahita Ghaedrahmati [0000-0002-5705-3555] | en |
| Issued Date | 2026-04-30 | en |
| Abstract | Background: Cryopreservation serves as a fundamental strategy for long-term preservation of genetic resources; however, the freeze-thaw process inevitably induces a range of structural and functional damage in spermatozoa. Quercetin, a flavonoid compound, has attracted considerable attention due to its potent antioxidant properties, and its potential protective effects against cryopreservation-induced oxidative damage have been widely investigated. Objectives: Given the documented positive effects of quercetin on sperm viability and the lack of consensus regarding its most appropriate concentration, this study aimed to assess the impact of different concentrations of quercetin (0, 5, 10, 20, and 40 µM) on apoptotic indices and reactive oxygen species (ROS) levels in thawed ram sperm prior to in vitro fertilization (IVF). Evaluations were performed at two post-thaw spermatozoa time points: 3 hours and 18 hours. Methods: Frozen ram sperm samples were rapidly thawed at 37 °C, washed, and then exposed to different concentrations of quercetin. The samples were incubated at 38.5 °C in an atmosphere containing 20% O2 and 5% CO2 for 3 and 18 hours. After incubation, apoptosis indices and ROS levels were measured using flow cytometry. Results: After 3 hours of incubation, quercetin caused a concentration-dependent modulation of ROS; the control group exhibited the highest ROS levels, while the 20 µM concentration (Group D) showed better regulation of intracellular oxidative status. The highest percentage of viable cells (34.2%) was observed in Group E (40 µM); however, a slight increase in ROS was observed at this concentration compared with 20 µM, suggesting a possible concentration-dependent variation in quercetin activity rather than a confirmed pro-oxidant effect. In contrast, after 18 hours of incubation, apoptosis and ROS levels showed less pronounced differences among groups, and the interpretation of ROS changes may be influenced by time-dependent alterations in sperm physiology. Overall, 20 - 40 µM may represent a potentially beneficial concentration range; however, further functional studies are required to determine the most appropriate dose. Conclusions: Quercetin treatment at 20 and 40 µM showed potential beneficial effects on oxidative status and sperm viability following thawing. However, considering the observed dose- and time-dependent responses, further functional studies are required to identify the optimal concentration and exposure conditions before its broader application in assisted reproductive research. | en |
| DOI | https://doi.org/10.5812/gct-173244 | en |
| URI | https://brieflands.com/journals/gct/articles/173244 | en |
| Keyword | Quercetin | en |
| Keyword | Oxidative Stress | en |
| Keyword | Apoptosis Index | en |
| Keyword | Flow Cytometry | en |
| Keyword | Antioxidant Activity | en |
| Publisher | Brieflands | en |
| Title | Dose- and Time-Dependent Effects of Quercetin on Apoptosis and Oxidative Stress in Post-Thaw Ram Spermatozoa | en |
| Type | Research Article | en |