Effects of β-sitosterol on Porcine Oocyte Maturation and Embryonic Development in Vitro
The aim of this study was to investigate the effects of β-sitosterol(SITO),a natural plant-derived antioxidant molecule,on the in vitro maturation efficiency of porcine oocytes and the quality of early embryo development in vitro.In this study,SITO with different concentra-tions were added into porcine oocytes in vitro maturation medium,which was the experimental group,and the medium without SITO(containing 0.16％DMSO)was the control group,the concentrations in experimental groups were 10,20 and 40 μmol·L-1 respectively,and they were used to culture the oocytes in vitro collected from porcine ovaries in slaughterhouse.Different concentrations were repeated 3 times,and 150-200 oocytes were cultured in each group.After 46 h of culture,measured the maturation rate of oocytes,and mature oocytes were collected for re-active oxygen species,apoptosis,mitochondrial membrane potential staining and qRT-PCR of re-lated genes;the adult oocytes were activated by parthenogenesis,the cleavage rate was calculated after 2 days,the blastocyst rate was calculated after 7 days,and the total number of blastocysts was calculated.The results showed that 20 μmol·L-1 SITO treatment significantly improved the maturation rate of porcine oocytes in vitro,promoted the formation of blastocysts after partheno-genetic activation,decreased the ROS in oocytes,and increased the expression of anti-oxidative stress genes SOD and CAT.At the same time,SITO decreased the apoptosis level of oocytes,increased the expression of anti-apoptotic gene BCL-2,and decreased the expression of pro-apop-totic genes Caspase 3 and BAX.In addition,SITO could enhance mitochondrial membrane po-tential(△Ψ m)and increase the expression of TFAM gene.In summary,the results of this study showed that adding 20 µmol·L-1 SITO to porcine oocyte in vitro maturation medium can significantly promote the rate of porcine oocytes,and improve the in vitro development ability and embryo quality.
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