Effect of resveratrol treatment on apoptosis and apoptotic pathways during boar semen freezing

Abstract

Resveratrol (3,5,4'-trihydroxystilbene, RSV) has been widely used in mammalian cells, but whether it can be used during freezing boar semen is still unknown. The effects of RSV treatment during boar semen freezing on its anti-freezing ability, apoptosis, and possible apoptotic pathways were observed in this study. Sperm motility, mitochondrial membrane potential (∆Ψ m), adenosine triphosphate (ATP) content, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL)-positive apoptotic state, and messenger RNA (mRNA) expression levels of apoptotic genes involved in different apoptotic pathways after freezing with or without RSV treatment were tested. The results showed that: (1) Compared with fresh sperm, the motility, normal acrosome rate, and plasma membrane integrity rate of frozen boar sperm decreased significantly (P<0.05), and RSV did not significantly increase the sperm motility (0.44 vs. 0.40, P>0.05), but it did significantly improve the normal acrosome rate (57.65% vs. 47.00%, P<0.05) and plasma membrane integrity rate (46.67% vs. 38.85%, P<0.05). (2) After freezing, most boar sperm showed low mitochondrial ∆Ψ m. RSV treatment could increase the rate of high mitochondrial ∆Ψ m of boar sperm. (3) RSV treatment significantly decreased reactive oxygen species (ROS) levels (58.65% vs. 88.41%, P<0.05) and increased the ATP content (0.49 μmol/L vs. 0.25 μmol/L, P<0.05) of boar sperm during freezing. (4) The apoptotic rate of the freezing group (80.41%) with TUNEL detection increased significantly compared to the fresh group (9.70%, P<0.05), and RSV treatment greatly decreased the apoptotic rate (68.32%, P<0.05). (5) Real-time polymerase chain reaction (RT-PCR) showed that not only the genes from the death receptor-mediated apoptotic pathway (tumor necrosis factor-α (TNF-α), Fas ligand (FasL), and Caspase-8), but also the genes from the mitochondria-mediated apoptotic pathway (manganese superoxide dismutase (MnSOD), B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and Caspase-9) were both significantly changed after freezing. RSV treatment during freezing greatly changed their expression levels. Although RSV treatment during boar semen freezing did not significantly increase motility after thawing, it still played an efficient antioxidant role, which could enhance the mitochondrial function and decrease the apoptotic level induced by both the death receptor-and mitochondria-mediated apoptotic pathways.

Resveratrol supplementation into extender protects against cryodamage in dog post-thaw sperm

“…The variation could be due to species differences, type of medium and buffer used, and emphasizes the need to optimize conditions when using RSV or other antioxidants in cryopreservation. Similar to our findings, the relatively high level of mitochondrial activity and integrity of the acrosome and chromatin in RSV observed in boar semen [ 19 ]. Moderate doses of RSV induced mitochondrial biogenesis and protected against a metabolic decline in mice [ 38 ].…”

“…The significant increase in the expression of BCL2 , a mitochondria-mediated apoptotic pathway gene, by RSV-supplementation in post-thaw dog sperm agreed with the findings in RSV-supplemented boar sperm [ 19 ] and rats [ 16 ]. This suggests that RSV improves the survival rate of sperm by reducing the stress-induced apoptotic pathways of cell death.…”

Effects of Resveratrol Supplementation on the Motility, Structural Integrity, and Mitochondrial Function of Freeze-Thawed Dog Sperm

“…The significant increase in the expression of BCL2, a mitochondria-mediated apoptotic pathway gene, by RSV-supplementation in post-thaw dog sperm agreed with the findings in RSV-supplemented boar sperm [53] and rats [54]. This suggests that RSV improves the survival rate of sperm by reducing the stress-induced apoptotic pathways of cell death.…”

“…The variation could be due to differences in species and the type of media and buffer used and emphasizes the need to optimize conditions when using RSV or other antioxidants in cryopreservation. The relatively high level of mitochondrial activity and integrity of the acrosome and chromatin in RSV-supplemented dog sperm was also observed in boar semen, in which enhanced mitochondrial function and decreased apoptosis, induced by both the death receptor-and mitochondria-mediated apoptotic pathways, were observed [37]. Moderate doses of RSV induced mitochondrial biogenesis and protected against metabolic decline in mice [38].…”

Antioxidants Significantly Improve Qinling Giant Panda (Ailuropoda Melanoleuca Qinlingensis) Sperm Quality During Cryopreservation

“…Finally, the semipermeable properties of the membrane are affected by peroxidative attack. However, the addition of RSV, LJP, and LBP to semen reduces the negative effects of excessive ROS levels [32][33][34][35]. Our study veri ed that supplementation of the freezing medium with RSV (50 µM), LBP (2.0 mg/mL), LJP (1.0 mg/mL) and combined addition signi cantly enhanced activity of SOD and GSH for oxygen radical (O2−) scavenging.…”