In vitro 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 cerrado 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 (Eugenia dysenterica) and murici (Byrsonima crassifolia) to be used during the in vitro culture of in vitro-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 in vitro cultivation. Such extracts may be used as alternatives to increase the cryotolerance of in vitro-produced embryos.

中文翻译：

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塞拉多生物群系干叶的乙醇提取物可提高牛体外产生的低温耐受性

体外胚胎产生（IVP）会引起活性氧（ROS）的过量产生，从而影响胚泡质量。因此，用抗氧化剂补充培养基是克服氧化应激损伤的替代方法。但是，越来越需要在细胞培养物中使用更天然，毒性更小的抗氧化剂。本研究旨在评估塞拉多叶片乙醇提取物对IVP的影响。首先，评估叶片的抗氧化能力和酚类化合物的量。然后，在体外使用的最佳乙醇提取物浓度由卡加塔（Eugenia dysenterica）和鼠李（Byrsonima crassifolia）组成。培养体外确定了产生的胚胎。之后，我们评估了两种植物提取物对ROS和谷胱甘肽（GSH）产生的影响，同时还评估了细胞凋亡和ROS代谢基因的表达。在随后的步骤中，研究了在胚胎培养过程中干燥的卡加塔咖啡和鼠李叶的乙醇提取物对膨胀胚泡的耐低温性的影响。结果显示，除了诱导GPX4和PRDX3转录水平增加外，用0.01 mg / mL的卡加塔乙醇提取物培养的胚胎中凋亡细胞的比例也大大降低。穆里西乙醇提取物诱导了这些基因的转录丰度增加，但并未降低凋亡细胞的比例。此外，用浓度为0的提取物培养的膨胀胚泡。与先前补充有提取物的冷冻组相比，01 mg / mL冷冻保存的孵化率更高，变性率更低。此外，冷冻保存12 h后培养的胚胎的凋亡率在以前接触过提取物的组中较低。体外培养。此类提取物可用作增加体外产生的胚胎的耐低温性的替代物。