Apigenin supplementation substantially improves rooster sperm freezability and post-thaw function

Abstract

This pioneering research investigated apigenin potential to augment rooster sperm cryosurvival in an extender model. Apigenin is a natural antioxidant flavonoid showing promise for improved post-thaw sperm function. However, its effects on avian semen cryopreservation remain unexplored. This first study supplemented rooster sperm Lake extender with 0, 50, 100, 200, 400 μmol/L apigenin to determine the optimal concentrations for post-thaw quality. Supplementation with 100 μmol/L apigenin resulted in significant enhancements in total motility (from 41.5% up to 71.5%), progressive motility (18.1% to 29.1%) (p < 0.05), membrane integrity (40% to 68%), mitochondrial function (p < 0.001), viability (37% to 62%) and total antioxidant capacity (p < 0.001) compared to the control. It also substantially reduced percentages of abnormal morphology, reactive oxygen species and apoptosis (p < 0.001). Although 200 μmol/L apigenin significantly enhanced some attributes, effects were markedly lower than 100 μmol/L. Higher doses did not improve cryoprotective parameters. This indicates 100 μmol/L as the optimal apigenin concentration. This represents the first report of apigenin protecting rooster sperm from cryodamage. The natural antioxidant improved post-thaw sperm quality, likely by suppressing oxidative stress and apoptosis. Apigenin shows promise for enhancing rooster sperm cryosurvival.

Keywords: Antioxidant; Apigenin; Cryopreservation; Rooster; Sperm.

Figure 1

TM total motility (a), PM progressive motility (b), VAP average path velocity (c) and VSL straight-line velocity (d) after thawing samples cryopreserved with different apigenin concentrations. Different letters indicate that treatments differ P < 0.05.

Figure 2

STR straightness (a), LIN linearity (b), ALH amplitude of the lateral head displacement (c) and BCF beat cross frequency (d) after thawing samples cryopreserved with different apigenin concentrations. Different letters indicate that treatments differ P < 0.05.

Figure 3

Membrane integrity (a), total abnormality (b), and mitochondrial activity (c) after thawing samples cryopreserved with different apigenin concentrations. Different letters indicate that treatments differ P < 0.05.

Figure 4

Viability (a), apoptotic (b) and dead sperm (c) after thawing samples cryopreserved with different apigenin concentrations. Different letters indicate that treatments differ P < 0.05.

Figure 5

ROS (a), GPx (b), SOD (c), and TAC (d) after thawing samples cryopreserved with different apigenin concentrations. Different letters indicate that treatments differ P < 0.05.