Shenzhen Key Laboratory of Reproductive Immunology for Peri-implantation, Shenzhen Zhongshan Institute for Reproductive Medicine and Genetics, Fertility Center, Shenzhen Zhongshan Urology Hospital, Shenzhen, China.
Biol Reprod. 2021 Aug 3;105(2):393-402. doi: 10.1093/biolre/ioab101.
Deterioration of the endometrial environment is an essential cause of recurrent miscarriage (RM). However, current studies in terms of endometrial amino acid metabolic characterization and autophagy are still inadequate. We tried to (1) identify the alternation in metabolite profiles in the RM endometrium; (2) investigate the expression of autophagy-related proteins in RM; and (3) elucidate the association between amino acid metabolism and autophagy in RM. Our results showed that glutamine metabolites were up-regulated in the endometrium of RM women. The levels of autophagy-associated proteins, LC3B, ATG12, and Beclin-1, were significantly higher in RM. Hemostasis, autophagy and IFNα signaling were the top three differentially activated signaling pathways between women with RM and normal pregnancy. Interestingly the expression of AMPK and GCN2 was significantly up-regulated in the endometrium of women with RM, and the same expression trend was also observed in the human endometrial stromal cells cultured in glutamine deprivation medium. Furthermore, inhibition of AMPK decreased the level of GCN2, indicating a positive correlation between GCN2 and AMPK. The expression of GCN2 was consistent with the expression of ATG12 and beclin-1; however, it was opposite to that of p62. Exposure to glutamine deprivation increased the level of LC3B, GCN2, ATG12, and beclin-1. Altogether, these findings suggested significant crosstalk between amino acid metabolism and autophagy. In summary, our data suggested that aberrant crosstalk between amino acid metabolism and autophagy may contribute to the impaired endometrial microenvironment of RM. Our study may provide new insight into the diagnosis of RM due to endometrial factors.
子宫内膜环境的恶化是复发性流产(RM)的一个重要原因。然而,目前关于子宫内膜氨基酸代谢特征和自噬的研究仍然不足。我们试图:(1)鉴定 RM 子宫内膜中代谢物谱的变化;(2)研究 RM 中自噬相关蛋白的表达;(3)阐明 RM 中氨基酸代谢与自噬之间的关系。我们的结果表明,RM 妇女的子宫内膜中谷氨酰胺代谢物上调。自噬相关蛋白 LC3B、ATG12 和 Beclin-1 的水平在 RM 中显著升高。止血、自噬和 IFNα 信号通路是 RM 患者和正常妊娠妇女之间差异激活的三个最重要的信号通路。有趣的是,RM 患者子宫内膜中 AMPK 和 GCN2 的表达显著上调,在缺乏谷氨酰胺的培养基中培养的人子宫内膜基质细胞中也观察到相同的表达趋势。此外,AMPK 的抑制降低了 GCN2 的水平,表明 GCN2 和 AMPK 之间存在正相关。GCN2 的表达与 ATG12 和 beclin-1 的表达一致;然而,它与 p62 的表达相反。暴露于缺乏谷氨酰胺的环境中会增加 LC3B、GCN2、ATG12 和 beclin-1 的水平。总之,这些发现表明氨基酸代谢和自噬之间存在显著的相互作用。综上所述,我们的数据表明,氨基酸代谢和自噬之间的异常相互作用可能导致 RM 子宫内膜微环境受损。我们的研究可能为基于子宫内膜因素的 RM 诊断提供新的见解。