Background: Asthenozoospermia (AZS), characterized by diminished sperm motility, represents a prevalent etiology of male infertility with multifactorial origins. While sperm cryo-preservation serves as a critical component of assisted repro- ductive technologies (ART), the freeze-thaw process induces excessive reactive oxygen species (ROS) generation, compro- mising sperm quality. This study evaluates the protective effects of theophylline - a phosphodiesterase (PDE) inhibitor exhibit- ing anti-inflammatory properties - on biochemical parameters during cryopreservation in asthenozoospermic men. Materials and Methods: Thirty semen samples were collected from men with asthenozoospermia referred to the Roya Infer- tility Treatment Center in Qom, Iran. Each semen sample was divided into three experimental groups: the control group (fresh sample), the freeze group (exposed to sperm freezing medium only), and the freeze + Theophylline group (treated with 0.05 mM theophylline). The levels of tumor necrosis factor alpha (TNF-α), interleukin-10 (IL-10), malondialdehyde (MDA), and antioxidant enzymes—superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX)—and total antioxidant capacity (TAC) were measured using the ELISA method. Results: The results showed a significant increase in TNF-α and MDA levels and a significant decrease in antioxidant en- zyme activity and IL-10 levels in the freezing group compared to the control group (P˂0.05). In contrast, a significant increase in the mean level of antioxidant enzymes and IL-10 and a sig- nificant decrease in MDA and TNF-α levels were observed in the freezing + theophylline group compared to the freezing group (P˂0.05). Conclusion: Our results suggest that the addition of theophyl- line to sperm cryopreservation media improves the biochemical profile and modulates inflammatory markers by reducing ROS in asthenozoospermic men.
