Multi-User Laboratory of Phytomedicines Pharmacology, and Biotechnology (PhitoPharmaTec), Department of Pharmacology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu 18618-000, Brazil.
Laboratory of Embryonic Micromanipulation, Department of Biological Sciences, School of Sciences and Languages, São Paulo State University (UNESP), Assis 19806-900, Brazil.
Int J Mol Sci. 2023 Sep 14;24(18):14060. doi: 10.3390/ijms241814060.
During embryo development, the endoplasmic reticulum (ER) acts as an important site for protein biosynthesis; however, in vitro culture (IVC) can negatively affect ER homeostasis. Therefore, the aim of our study was to evaluate the effects of the supplementation of tauroursodeoxycholic acid (TUDCA), an ER stress inhibitor, in the IVC of bovine embryos. Two experiments were carried out: Exp. 1: an evaluation of blastocyst rate, hatching kinetics, and gene expression of hatched embryos after being treated with different concentrations of TUDCA (50, 200, or 1000 μM) in the IVC; Exp. 2: an evaluation of the re-expansion, hatching, and gene expression of hatched embryos previously treated with 200 µM of TUDCA at IVC and submitted to vitrification. There was no increase in the blastocyst and hatched blastocyst rates treated with TUDCA in the IVC. However, embryos submitted to vitrification after treatment with 200 µM of TUDCA underwent an increased hatching rate post-warming together with a down-regulation in the expression of ER stress-related genes and the accumulation of lipids. In conclusion, this work showed that the addition of TUDCA during in vitro culture can improve the cryotolerance of the bovine blastocyst through the putative modulation of ER and oxidative stress.
在胚胎发育过程中，内质网（ER）是蛋白质生物合成的重要场所；然而，体外培养（IVC）会对 ER 稳态产生负面影响。因此，本研究旨在评估牛胚胎 IVC 中添加牛磺熊脱氧胆酸（TUDCA）作为 ER 应激抑制剂的效果。进行了两项实验：实验 1：评估不同浓度 TUDCA（50、200 或 1000 μM）处理对囊胚率、孵化动力学和孵化胚胎基因表达的影响；实验 2：评估 200 μM TUDCA 处理的 IVC 后胚胎的再扩张、孵化和孵化胚胎基因表达，并进行玻璃化处理。TUDCA 在 IVC 中处理并未增加囊胚和孵化囊胚的形成率。然而，经 200 μM TUDCA 处理后进行玻璃化处理的胚胎在解冻后孵化率增加，同时与 ER 应激相关基因表达下调和脂质积累相关。总之，这项工作表明，在体外培养过程中添加 TUDCA 可以通过可能调节 ER 和氧化应激来提高牛囊胚的抗冻能力。
