Veterinary Department, University Center of Brasília-UniCeub, Brasília, Brazil.
School of Agriculture and Veterinary Medicine, University of Brasília, Brazil.
Oxid Med Cell Longev. 2020 Nov 22;2020:6046013. doi: 10.1155/2020/6046013. eCollection 2020.
embryo production (IVP) induces excessive production of reactive oxygen species (ROS), which affects blastocyst quality. Therefore, the supplementation of culture media with antioxidants is an alternative to overcome oxidative stress damage. However, there is a growing demand for the use of antioxidant compounds that are more natural and less toxic in cell cultures. The present study is aimed at evaluating the effect of ethanolic extracts from leaves on IVP. First, the antioxidant capacity and the amount of phenolic compounds of the leaves were evaluated. Then, the best ethanolic extract concentration composed of cagaita () and murici () to be used during the culture of -produced embryos was determined. Afterward, we evaluated the influence of the extract of both plants on ROS and glutathione (GSH) production, while also evaluating the apoptosis and ROS metabolism gene expression. In a subsequent step, the effect of the ethanolic extracts of dried cagaita and murici leaves during embryonic cultivation on the cryotolerance of expanded blastocysts was studied. The results showed a significant reduction in the proportion of apoptotic cells from embryos cultivated with 0.01 mg/mL of the cagaita ethanolic extract, besides inducing an increase in the GPX4 and PRDX3 transcription levels. The murici ethanolic extract induced an increase in the transcription abundance of these genes but did not reduce the proportion of apoptotic cells. In addition, expanded blastocysts cultivated with extracts at a concentration of 0.01 mg/mL and cryopreserved had higher hatching rates and lower degeneration rates when compared to the frozen group previously supplemented with the extracts. Moreover, the apoptosis rate of embryos cultured for 12 h after cryopreservation was lower in groups previously exposed to extracts during cultivation. Such extracts may be used as alternatives to increase the cryotolerance of -produced embryos.
胚胎生产(IVP)会导致活性氧(ROS)的过度产生,这会影响囊胚的质量。因此,在培养基中添加抗氧化剂是克服氧化应激损伤的一种替代方法。然而,人们越来越需要使用在细胞培养中更天然、毒性更小的抗氧化化合物。本研究旨在评估叶乙醇提取物对 IVP 的影响。首先,评估了叶的抗氧化能力和酚类化合物的含量。然后,确定了由 cagaita()和 murici()组成的最佳乙醇提取物浓度,用于-产生胚胎的培养。之后,我们评估了两种植物的提取物对 ROS 和谷胱甘肽(GSH)产生的影响,同时还评估了细胞凋亡和 ROS 代谢基因表达。在随后的步骤中,研究了在胚胎培养过程中使用干 cagaita 和 murici 叶的乙醇提取物对扩展囊胚的抗冻能力的影响。结果表明,与用 0.01 mg/mL cagaita 乙醇提取物培养的胚胎相比,凋亡细胞的比例显著降低,同时 GPX4 和 PRDX3 的转录水平增加。murici 乙醇提取物诱导这些基因的转录丰度增加,但不降低凋亡细胞的比例。此外,与先前用提取物补充的冷冻组相比,用提取物在 0.01 mg/mL 浓度下培养的扩展囊胚具有更高的孵化率和更低的退化率。此外,与冷冻组相比,在冷冻前暴露于提取物的胚胎在冷冻后培养 12 小时的凋亡率更低。这些提取物可作为提高-产生胚胎抗冻能力的替代方法。