Embelin supplementation of in vitro maturation medium does not influence nuclear and cytoplasmic maturation of pig oocytes

Oxidative stress caused from in vitro culture contributes to inadequate oocyte maturation which leads to a poor embryo development. Therefore,it is important to protect oocytes and embryos against oxidative stress. This study was aimed at evaluating the effect of Embelin (2,5-dihydroxy-3-undecyl-1,4-benzoquinone),an antioxidant with various pharmacologic activities,on nuclear and cytoplasmic maturation of pig oocytes as well as on steroidogenesis of cumulus cells (CCs). Another objective was to determine the influence of Embelin on developmental competence of pig oocytes as well as the expression levels of three key genes (Nanog,Sox2 and Oct4) involved in the control of pluripotency in parthenogenetically activated embryos. Embelin (0,10,20 and 40 µM) was added during in vitro maturation of cumulus oocyte complexes; media of both the first and the second day of culture were collected and assayed for progesterone and estradiol-17β. At the end of the maturation period,the oocytes were fixed (to determine nuclear maturation) or partenogenically activated to evaluate cytoplasmic maturation and genes expression. Embelin did not exert any effect on the proportion of MII oocytes,steroidogenesis of CCs,percentage of embryos that developed to blastocyst stage and the number of blastomeres/blastocyst. Moreover,no significant differences of Oct4,Nanog and Sox2 transcripts were detected in blastocyst stage embryos. In conclusion,Embelin did not influence the reproductive parameters assessed,confirming that it is not possible to predict whether the beneficial effect exerted by an antioxidant in a particular tissue could be present also in another one.