School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai Key Laboratory of Veterinary Biotechnology , Shanghai , China.
Division of Animal and Nutritional Sciences, West Virginia University , Morgantown, West Virginia.
Am J Physiol Cell Physiol. 2019 Feb 1;316(2):C198-C209. doi: 10.1152/ajpcell.00256.2018. Epub 2018 Nov 28.
Significant embryo loss remains a serious problem in pig production. Reactive oxygen species (ROS) play a critical role in embryonic implantation and placentation. However, the potential mechanism of ROS on porcine trophectoderm (pTr) cell fate during the peri-implantation period has not been investigated. This study aimed to elucidate the effects of ROS on pTr cell phenotypes and the regulatory role in cell attachment and differentiation. Herein, results showed that exogenous HO inhibited pTr cell viability, arrested the cell cycle at S and G2/M phases, and increased cell apoptosis and autophagy protein light chain 3B and Beclin-1, whereas these effects were reversed by different concentrations of N-acetyl-l-cysteine (NAC) posttreatment. In addition, NAC abolished HO-induced autophagic flux, inhibited intracellular and mitochondrial ROS, and restored expression of genes important for mitochondrial DNA and biogenesis, cell attachment, and differentiation. NAC reversed HO-activated MAPK and Akt/mammalian target of rapamycin pathways in dose-dependent manners. Furthermore, analyses with pharmacological and RNA interference approaches suggested that autophagy regulated cell apoptosis and gene expression of caudal-related homeobox 2 and IL-1β. Collectively, these results provide new insights into the role of the ROS-induced autophagy in pTr cell apoptosis, attachment, and differentiation, indicating a promising target for decreasing porcine conceptus loss during the peri-implantation period.
胚胎丢失仍然是养猪生产中的一个严重问题。活性氧（ROS）在胚胎着床和胎盘形成中起着关键作用。然而，ROS 对围植入期猪滋养层（pTr）细胞命运的潜在机制尚未得到研究。本研究旨在阐明 ROS 对 pTr 细胞表型的影响及其对细胞黏附和分化的调控作用。结果表明，外源性 HO 抑制 pTr 细胞活力，将细胞周期阻滞在 S 和 G2/M 期，并增加细胞凋亡和自噬蛋白 LC3B 和 Beclin-1，而 N-乙酰-L-半胱氨酸（NAC）后处理可逆转这些作用。此外，NAC 消除了 HO 诱导的自噬流，抑制了细胞内和线粒体 ROS，并恢复了与线粒体 DNA 和生物发生、细胞黏附和分化有关的基因的表达。NAC 以剂量依赖的方式逆转了 HO 激活的 MAPK 和 Akt/哺乳动物雷帕霉素靶蛋白途径。此外，通过药理学和 RNA 干扰方法的分析表明，自噬调节细胞凋亡和尾相关同源盒 2 和白细胞介素 1β的基因表达。综上所述，这些结果为 ROS 诱导的自噬在 pTr 细胞凋亡、黏附和分化中的作用提供了新的见解，表明这是减少围植入期猪胚胎丢失的有希望的靶点。
