Enhancing Male Fertility: The Role of Crocin in Boosting Sperm Motility Through Antioxidant Activity and Mitochondrial Pathways

Abstract

Excessive production of reactive oxygen species (ROS) is a critical factor adversely affecting semen quality, particularly sperm motility. Crocin, a key compound of Crocus Sativus (saffron), is recognized for its antioxidant properties. This study aimed to investigate the potential of crocin to improve in vitro sperm motility in case of asthenozoospermia. Sperm samples from 95 patients with asthenozoospermia were incubated for 3 h at 37°C with varying concentrations of crocin (0, 0.2, 0.5, 1, or 1.5 mM). Sperm motility was assessed using time-lapse video microscopy. Intracellular ROS levels were evaluated through the DCFH-DA assay while ROS-induced damage was quantified through lipid peroxidation and carbonylated proteins levels. Catalase, superoxide dismutase, and glutathione peroxidase were analyzed. Additionally, mitochondrial function was assessed via mitochondrial activity via MTT assay and potential via Rh123 assay. Our findings revealed a significant increase in sperm velocity by 93% (G1) and 91% (G2) and in the distance traveled by motile spermatozoa by 76.5% (G1) and 72.1% (G2) (p < 0.001). ROS levels decreased by 1.5-fold (G1) and 2.4-fold in (G2) (p < 0.001), while malondialdehyde levels dropped twofold (G1) and 3.4-fold (G2) (p < 0.001). Carbonylated protein content decreased by 2.8-fold (G1) and threefold (G2) (p < 0.001). Improvements also included succinate dehydrogenase activity (2.14-fold in G1, 1.85-fold in G2, p < 0.001) and mitochondrial membrane potential (1.7-fold in G1, 1.9-fold in G2, p < 0.001). These findings highlight crocin's potential to enhance sperm motility and reduce oxidative stress in asthenozoospermic samples.