Abstract：
BACKGROUND: Clinically, recurrent spontaneous abortion (RSA) is a pregnancy illness that is difficult to treat. Impaired decidualization is a documented cause of RSA, but the etiology and mechanism are still unknown. cAMP-responsive element binding protein 5 (CREB5) is a member of the ATF/CREB family. CREB5 has been reported to be related to pathological pregnancy, but there are few related studies on this topic in patients with RSA, and the underlying mechanism is unclear.
METHODS: We collected decidual tissues from RSA patients and healthy pregnant women to measure the expression level of CREB5, PRL, IGFBP1, ATG5, LC3B, and SQSTM/p62. Then, the changes in CREB5 expression and autophagy levels were measured in human endometrial stromal cells (hESCs) during decidualization. The expression levels of PRL and IGFBP1 were tested in sh-CREB5/ov-CREB5 hESCs after decidualization induction, and the autophagy level in sh-CREB5/ov-CREB5 hESCs was measured without decidualization induction. The decidualization ability of sh-CREB5 and ov-CREB5 hESCs treated with an autophagy inducer or inhibitor was measured. To investigate the effect of CREB5 in hESCs on the invasion and migration of HTR8/SVneo cells, we performed a coculture experiment. Finally, we examined the expression of CREB5 and autophagy key proteins in mouse decidual tissues by constructing an abortion mouse model.
RESULTS: In our study, we found that the expression of CREB5 was unusually elevated in the uterine decidua of RSA patients, but the expression of PRL, IGFBP1, and autophagy were decreased. During the decidualization of hESCs, the expression of CREB5 gradually decreases in a time-dependent manner with increasing autophagy. Moreover, by knocking down or overexpressing CREB5 in hESCs, it was found that CREB5 can impair decidualization and reduce autophagy in hESCs. Furthermore, the damage caused by CREB5 in terms of decidualization can be reversed by the addition of an autophagy inducer (rapamycin). In addition, CREB5 can increase the secretion of proteins (IL-1β and TGF-β1) in hESCs to inhibit trophoblast invasion and migration.
CONCLUSIONS: Our data support the supposition that CREB5 disturbs the decidualization of endometrial stromal cells and interactions at the maternal-fetal interface by inhibiting autophagy and that its abnormal upregulation and dysfunction may lead to RSA. It may function as a diagnostic and therapeutic target for RSA. Similarly, we found that in the spontaneous abortion mouse model, the expression of CREB5 in the decidual tissue of the abortion group was significantly increased, and autophagy was decreased.
METHODS: We collected decidual tissues from RSA patients and healthy pregnant women to measure the expression level of CREB5, PRL, IGFBP1, ATG5, LC3B, and SQSTM/p62. Then, the changes in CREB5 expression and autophagy levels were measured in human endometrial stromal cells (hESCs) during decidualization. The expression levels of PRL and IGFBP1 were tested in sh-CREB5/ov-CREB5 hESCs after decidualization induction, and the autophagy level in sh-CREB5/ov-CREB5 hESCs was measured without decidualization induction. The decidualization ability of sh-CREB5 and ov-CREB5 hESCs treated with an autophagy inducer or inhibitor was measured. To investigate the effect of CREB5 in hESCs on the invasion and migration of HTR8/SVneo cells, we performed a coculture experiment. Finally, we examined the expression of CREB5 and autophagy key proteins in mouse decidual tissues by constructing an abortion mouse model.
RESULTS: In our study, we found that the expression of CREB5 was unusually elevated in the uterine decidua of RSA patients, but the expression of PRL, IGFBP1, and autophagy were decreased. During the decidualization of hESCs, the expression of CREB5 gradually decreases in a time-dependent manner with increasing autophagy. Moreover, by knocking down or overexpressing CREB5 in hESCs, it was found that CREB5 can impair decidualization and reduce autophagy in hESCs. Furthermore, the damage caused by CREB5 in terms of decidualization can be reversed by the addition of an autophagy inducer (rapamycin). In addition, CREB5 can increase the secretion of proteins (IL-1β and TGF-β1) in hESCs to inhibit trophoblast invasion and migration.
CONCLUSIONS: Our data support the supposition that CREB5 disturbs the decidualization of endometrial stromal cells and interactions at the maternal-fetal interface by inhibiting autophagy and that its abnormal upregulation and dysfunction may lead to RSA. It may function as a diagnostic and therapeutic target for RSA. Similarly, we found that in the spontaneous abortion mouse model, the expression of CREB5 in the decidual tissue of the abortion group was significantly increased, and autophagy was decreased.
摘要：
背景：临床上，复发性自然流产（RSA）是一种难以治疗的妊娠疾病。判定化受损是RSA的一个记录在案的原因，但病因和机制尚不清楚。cAMP反应元件结合蛋白5(CREB5)是ATF/CREB家族的成员。据报道，CREB5与病理性妊娠有关，但是在RSA患者中很少有相关的研究，潜在机制尚不清楚。
方法：我们收集RSA患者和健康孕妇的蜕膜组织，以测量CREB5，PRL，IGFBP1,ATG5,LC3B,和SQSTM/p62。然后，在蜕膜化过程中测量人子宫内膜基质细胞(hESCs)中CREB5表达和自噬水平的变化.在蜕膜化诱导后的sh-CREB5/ov-CREB5hESCs中检测PRL和IGFBP1的表达水平,并且在没有蜕膜化诱导的情况下测量sh-CREB5/ov-CREB5hESCs中的自噬水平。测量用自噬诱导剂或抑制剂处理的sh-CREB5和ov-CREB5hESC的蜕膜化能力。探讨hESCs中CREB5对HTR8/SVneo细胞侵袭和迁移的影响,我们进行了共培养实验。最后,通过构建流产小鼠模型,检测CREB5和自噬关键蛋白在小鼠蜕膜组织中的表达。
结果：在我们的研究中，我们发现CREB5的表达在RSA患者的子宫蜕膜中异常升高，但是PRL的表达，IGFBP1和自噬均降低。在hESC的蜕膜化过程中，随着自噬的增加,CREB5的表达呈时间依赖性逐渐降低。此外，通过在hESC中敲低或过表达CREB5，发现CREB5可以损害hESCs的蜕膜化和减少自噬。此外，CREB5在蜕膜化方面引起的损伤可以通过添加自噬诱导剂（雷帕霉素）来逆转。此外,CREB5可以增加hESCs中蛋白质（IL-1β和TGF-β1）的分泌，从而抑制滋养细胞的侵袭和迁移。
结论：我们的数据支持以下假设：CREB5通过抑制自噬干扰子宫内膜基质细胞的蜕膜化和母胎界面的相互作用，其异常的上调和功能障碍可能导致RSA。它可以作为RSA的诊断和治疗靶标。同样，我们发现在自然流产小鼠模型中,流产组蜕膜组织中CREB5的表达显著升高,自噬减少。
方法：我们收集RSA患者和健康孕妇的蜕膜组织，以测量CREB5，PRL，IGFBP1,ATG5,LC3B,和SQSTM/p62。然后，在蜕膜化过程中测量人子宫内膜基质细胞(hESCs)中CREB5表达和自噬水平的变化.在蜕膜化诱导后的sh-CREB5/ov-CREB5hESCs中检测PRL和IGFBP1的表达水平,并且在没有蜕膜化诱导的情况下测量sh-CREB5/ov-CREB5hESCs中的自噬水平。测量用自噬诱导剂或抑制剂处理的sh-CREB5和ov-CREB5hESC的蜕膜化能力。探讨hESCs中CREB5对HTR8/SVneo细胞侵袭和迁移的影响,我们进行了共培养实验。最后,通过构建流产小鼠模型,检测CREB5和自噬关键蛋白在小鼠蜕膜组织中的表达。
结果：在我们的研究中，我们发现CREB5的表达在RSA患者的子宫蜕膜中异常升高，但是PRL的表达，IGFBP1和自噬均降低。在hESC的蜕膜化过程中，随着自噬的增加,CREB5的表达呈时间依赖性逐渐降低。此外，通过在hESC中敲低或过表达CREB5，发现CREB5可以损害hESCs的蜕膜化和减少自噬。此外，CREB5在蜕膜化方面引起的损伤可以通过添加自噬诱导剂（雷帕霉素）来逆转。此外,CREB5可以增加hESCs中蛋白质（IL-1β和TGF-β1）的分泌，从而抑制滋养细胞的侵袭和迁移。
结论：我们的数据支持以下假设：CREB5通过抑制自噬干扰子宫内膜基质细胞的蜕膜化和母胎界面的相互作用，其异常的上调和功能障碍可能导致RSA。它可以作为RSA的诊断和治疗靶标。同样，我们发现在自然流产小鼠模型中,流产组蜕膜组织中CREB5的表达显著升高,自噬减少。
