earticle

영어

In vitro production of pig embryos has been emerged for xenotransplantation due to physiological properties. Sirtuins play a crucial role in several cellular processes including DNA repair, apoptosis, cell cycle, and determining lifespan as NAD+ dependent class III histone deacetylase. Previous studies have shown that sirtuin inhibition induces embryonic developmental arrest and oxidative stress in porcine and murine models. Also, sirtuins regulate autophagy pathways through the deacetylation of ATG proteins and transcriptional factors. However, sirtuins-mediated mechanisms have not been examined in blastocysts. Therefore, we investigated the relationship between sirtuin inhibition and autophagy in porcine model. Embryos were cultured in 100 μM sirtinol (SIRT1/2 target inhibitor)-treated NCSU-23 media after in vitro fertilization. Treatment with sirtinol significantly decreased the rates of morula (21.34±1.84 vs. 11.89±2.01%), blastocyst formation (17.18±1.81 vs. 9.00±2.02%), and total cell number (50.80±1.47 vs. 37.71±1.79). Especially, expanded blastocysts (9.90±1.56 vs. 2.92±0.94%) were barely observed in sirtinol treated group. The level of Sirt2 expression was significantly decreased in sirtinol-treated group compared to control. Also, sirtinol induced autophagy by increasing LC3 transcript and LC3 protein in treatment group. Beclin1 and ATG5 expressions showed a slight increase in sirtinol-treated group. Finally, sirtuin inhibition by sirtinol showed significantly increased TUNEL indices (6.55±0.84 vs. 11.44±0.81) and fragmentation indices (0.33±0.05 vs. 1.40±0.30). Similarly, the level of BCL2L1 was lower, while the level of Caspase-3 expression was significantly elevated in the treated group compared to controls. Therefore, these results suggest that sirtuin inhibition may play an important role in porcine preimplantation development and embryo quality through regulation of autophagy and apoptosis pathways.