|
چکیده
|
Background: Asthenozoospermia, characterized by reduced or absent sperm motility, is a major cause of male infertility. Sperm cryopreservation, a vital technique in assisted reproductive technologies, induces cryoinjury, compromising sperm structure and function through oxidative stress, DNA damage, and apoptosis. Excessive reactive oxygen species (ROS) production during freeze-thaw cycles exacerbates oxidative stress, particularly in asthenozoospermic men, impairing semen quality and fertilization potential. Melatonin, a potent antioxidant, may mitigate these effects. Objective: This study aimed to evaluate the effects of melatonin, on sperm biochemical parameters and intracellular ROS levels in cryopreserved samples from asthenozoospermic men. Materials and Methods: This experimental in vitro study involved semen samples from 30 asthenozoospermic men, collected at the Amir-AL- Momenin Infertility Center, Arak, Iran from November 2023 to Jun 2024. Samples were divided into 3 groups: control (fresh), freeze, and freeze + melatonin (treated with cryoprotectant + 1 mM melatonin). Samples in the freeze groups were cryopreserved using a human sperm freezing medium and rapid freezing protocol. Levels of malondialdehyde (MDA), total antioxidant capacity, and antioxidant enzymes (catalase, glutathione, superoxide dismutase) were quantified using enzyme- linked immunosorbent assay. Intracellular ROS levels were measured using the DCFH-DA fluorescent probe via a Partec Pas flow cytometer with a 488 nm argon laser. Results: The mean levels of sperm antioxidant enzymes and total antioxidant capacity in the freezing group were significantly reduced compared to the control group (p < 0.001). In contrast, the mean levels of sperm MDA and ROS in the freezing group showed a significant increase (p < 0.001). A significant increase in the sperm antioxidant enzymes and TAC levels (p < 0.001), along with a significant decrease in MDA (p < 0.001) and ROS (p = 0.005) levels were seen in the freeze + melatonin group compared to the freezing group. Conclusion: In conclusion, incorporating melatonin into the freezing medium significantly reduces the adverse effects of cryopreservation on the sperm of asthenozoospermic men by mitigating oxidative stress and lowering ROS levels. This protective role of melatonin highlights its potential as an effective antioxidant in improving the quality and functionality of cryopreserved sperm, ultimately contributing to better outcomes in assisted reproductive technology.
|