Effects of riboflavin on boar sperm motility, sperm quality, enzyme activity and antioxidant status during cryopreservation

Abstract

Objectives This study was conducted to evaluate the effect of adding riboflavin to boar sperm freezing extender on the challenge of cryopreservation. Methods Different concentrations (0, 5, 10, 15, 20 or 25 μM) of riboflavin were added to the freezing extender. Spermatozoa motility, membrane integrity, acrosomal integrity, mitochondrial membrane potential and enzyme activities were analysed once 10 min after thawing. Q‐PCR was used to detect the mRNA expression of Caspase3, Bcl‐2 and Bax. Results The results showed that the addition of 10 μM riboflavin to boar sperm freezing extender significantly increased the frozen‐thawed sperm progressive motility compared with the control group (p < 0.05). Activities of superoxide dismutase, glutathione peroxidase and catalase improved after adding riboflavin to the extender (p < 0.05). During freezing‐thawing, the boar sperm mitochondrial membrane potential, acrosomal integrity, plasma membrane and DNA at 10 μM in the riboflavin group increased by 6.6%, 9.6%, 5.49% and 5.62% (p < 0.05), respectively, compared with the control group. The addition of 10 μM riboflavin to the extender significantly decreased the malondialdehyde (p < 0.05) content, whereas it increased the ATP content (p < 0.05) of boar sperm during freezing‐thawing. Furthermore, the expression of Caspase‐3 and Bax (p < 0.05) were significantly lower, whereas the expression of BCL‐2 (p < 0.05) was greater than the control group when adding 10 μM riboflavin to the extender. Conclusions Riboflavin showed cryoprotective capacity to the freezing extender used for boar sperm during the process of freezing‐thawing, and the optimal concentration of riboflavin for the frozen extender was 10 μM.

Antioxidant Effects of Myo-Inositol Improve the Function and Fertility of Cryopreserved Boar Semen

“…During cryopreservation, an increase in free radicals leads to oxidative stress, which leads to structural and functional changes and finally apoptosis. Myo-inositol improves sperm quality, reduces ROS production, protects acrosome integrity and mitochondrial function, protects against cryodamage, and reduces apoptosis [ 40 , 41 , 42 , 43 ].…”

Phosphocreatine addition to extender enhances the quality and antioxidant capacity of cryopreserved boar sperm

“…Sperm are produced in supraphysiological doses of ROS during cryopreservation and thawing, which leads to reduced sperm motility and viability, and increases the risk of DNA damage (Khosravizadeh et al, 2022). Numerous studies in recent years have shown that the exogenous antioxidants, including butylated hydroxytoluene, reduced glutathione (de Andrade et al, 2023), riboflavin (Dong et al, 2022), melatonin (Pezo et al, 2021), astaxanthin (Guo et al, 2021), rutin (Xu et al, 2020), resveratrol (Zhu, Li, et al, 2019) and alpha lipoic acid (Shen et al, 2016) have been applied to extenders to minimize the level of oxidative stress caused by ROS to boar sperm during cryopreservation.…”