Abstract
The Splinter bean, Entada abyssinica, is widely used in folk medicine. In the current work, we profiled the secondary metabolites from E. abyssinica bark extract using LC-MS and investigated its effect on cryopreserved ram semen. Twenty-eight compounds, including tannins and gallic acid derivatives that prevailed in the extract, were tentatively identified. Results showed that the quality of the post-thawed semen showed a significant improvement when the extract was added to the extender at a concentration of 375 μg/mL. The progressive motility and plasma membrane integrity of sperm cells were significantly increased in the post-thawed semen; however, the total antioxidant capacity (TAC) was insignificantly increased. A significant decrease in the concentration of hydrogen peroxide was detected as well. No significant changes were observed in activities of lactate dehydrogenase (LDH), alanine aminotransaminase (ALT), and aspartate transaminase (AST) within the treated samples. Intact sperm percentage was significantly increased, while apoptotic and necrotic sperm percentages were reduced significantly. Molecular docking of some individual components from the extract revealed their potential to interfere with the apoptosis cascade in which Bcl-2 is involved. In conclusion, Entada abyssinica appears to be useful for cryopreservation presumably owing to its polyphenol content that has potent antioxidant capacity scavenging reactive oxygen species (ROS), enhancing the endogenous antioxidant system and inhibiting lipid peroxidation.
Highlights
Artificial insemination (AI) is a widely applied technique that uses fresh semen and frozen–thawed sperms
28 secondary metabolites were tentatively identified in the methanol extract from E. abyssinica bark based on their molecular weight, mass fragmentation pattern, available authentic compounds, in-house library, and online literature
Previous studies reported that sperm viability and motility, the integrity of both plasma membrane and acrosome in postthawed semen are negatively affected during the cryopreservation process [33, 34]
Summary
Artificial insemination (AI) is a widely applied technique that uses fresh semen and frozen–thawed sperms. The use of ram frozen–thawed semen eliminates the geographical barriers, helps in preserving endangered breeds, and conserves the biodiversity [3]. Ice crystallization and recrystallization during freezing and thawing techniques induce biochemical and cellular changes and alter the sperm efficiency [7]. Sperm motility and morphology may be affected as well by increased membrane permeability after cryopreservation [8, 9]. These post-thawing-induced changes could impair sperm transport and survival inside the female reproductive tract, affecting fertilizing capacity and embryogenesis [8]. Many efforts to maintain and improve sperm viability in these techniques have been developing recently
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