Astaxanthin Supplementation Improves the Subsequent Developmental Competence of Vitrified Porcine Zygotes

Abstract

Cryopreservation of embryos has been confirmed to cause oxidative stress as a factor responsible for impaired developmental competence. Currently, astaxanthin (Ax) raises considerable interest as a strong exogenous antioxidant and for its potential in reproductive biology. The present study aimed to investigate the beneficial effects of Ax supplementation during in vitro culture of vitrified porcine zygotes and the possible underlying mechanisms. First, the parthenogenetic zygotes were submitted to vitrification and then cultured in the medium added with various concentrations of Ax (0, 0.5, 1.5, and 2.5 μM). Supplementation of 1.5 μM Ax achieved the highest blastocyst yield and was considered as the optimal concentration. This concentration also improved the blastocyst formation rate of vitrified cloned zygotes. Moreover, the vitrified parthenogenetic zygotes cultured with Ax exhibited significantly increased mRNA expression of CDX2, SOD2, and GPX4 in their blastocysts. We further analyzed oxidative stress, mitochondrial and lysosomal function in the 4-cell embryos and blastocysts derived from parthenogenetic zygotes. For the 4-cell embryos, vitrification disturbed the levels of reactive oxygen species (ROS) and glutathione (GSH), and the activities of mitochondria, lysosome and cathepsin B, and Ax supplementation could fully or partially rescue these values. The blastocysts obtained from vitrified zygotes showed significantly reduced ATP content and elevated cathepsin B activity, which also was recovered by Ax supplementation. There were no significant differences in other parameters mentioned above for the resultant blastocysts. Furthermore, the addition of Ax significantly enhanced mitochondrial activity and reduced lysosomal activity in resultant blastocysts. In conclusion, these findings revealed that Ax supplementation during the culture period improved subsequent embryonic development and quality of porcine zygotes after vitrification and might be used to ameliorate the recovery culture condition for vitrified embryos.

Astaxanthin Added during Post-Warm Recovery Mitigated Oxidative Stress in Bovine Vitrified Oocytes and Improved Quality of Resulting Blastocysts

“…This indicates that increased lysosomal activity can be associated with induced autophagy as a survival response. Similar trends in mitochondrial and lysosomal activities were observed in porcine four-cell embryos and blastocysts after astaxanthin addition [42].…”

“…It is more likely that effects of AX on oocyte and embryo development vary depending on the concentration used. Thus, [42] described the accelerated formation of blastocysts from porcine zygotes vitrified after parthenogenesis as well as higher blastocyst rate at the concentration of 1.5 µM AX. Increased blastocyst rate after astaxanthin addition was also obtained in aging porcine oocytes [26].…”

Astaxanthin promotes locomotor function recovery and attenuates tissue damage in rats following spinal cord injury: a systematic review and trial sequential analysis

“…Compared with the controls, rats that underwent AST treatment manifested increased levels of Beclin1 and LC3 and a decreased expression of p62 ( Abbaszadeh et al, 2022 ). Additionally, AST was shown to restore the normal function of lysosomes and notably inhibited the activation of mTOR complex1 ( Fu et al, 2022 ; Xiang et al, 2022 ), while this mechanism remains to be confirmed in SCI models.…”

Resveratrol Reduces ROS by Increasing GSH in Vitrified Sheep Embryos

“…It could also be because it increases the concentration of intracellular reduced glutathione (GSH) and favors the concentrations of the enzyme glutathione peroxidase, essential for the embryonic oxidant balance [4]. This allows a decrease in ROS production, as has been observed in pigs, cattle, sheep, and goats [2,3,[5][6][7][8][9][10][11].…”

“…Other antioxidants have been proven in porcine experiments, where Xiang et al [10] found that when adding 1.5 µM of astaxanthin to the culture medium the developmental competence of parthenogenetic zygotes improved (51.7% blastocysts). This concentration also improved the blastocyst formation rate of vitrified cloned zygotes (23.3% blastocysts).…”