Gao Peng, Zha Ying, Wei Lijie, Zhou Xuan, Zhu Shenglan, Zhang Huiting, Gao Xuan, Jiang Yi, Chen Yuting, Li Jiaqi, Zhang Jingyi, Yu Jun, Wang Shaoshuai, Liu Haiyi, Feng Ling
Department of Gynecology and Obstetrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
Department of Gynecology and Obstetrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
Placenta. 2022 Apr;121:164-172. doi: 10.1016/j.placenta.2022.03.125. Epub 2022 Mar 26.
The etiology of approximately half of patients with recurrent spontaneous abortion (RSA) has yet to be established. Granulocyte-colony stimulating factor (G-CSF) exerts a protective effect on pregnancy and its absence may lead to pregnancy failure. However, the effects and mechanisms of G-CSF activities have not been fully explored. Therefore, we aimed at evaluating whether a loss of G-CSF induces RSA by affecting cell communication at the maternal-foetal interface.
Villous and decidual tissues were obtained from participants and expression levels of G-CSF determined by qRT-PCR, Western blot and immunohistochemistry. G-CSF levels in trophoblasts were downregulated by siRNA. Exosomes were extracted from trophoblasts and co-cultured with macrophages. Molecular expression levels of key genes were determined by qRT-PCR and Western blot. Migration and proliferation of cells were evaluated by transwell and CCK8 assays. The RSA mice models were intraperitoneally administered with G-CSF to assess pregnancy outcomes and expression profiles of G-CSF as well as its receptor at the mother-foetal interface.
Relative to the decidua, G-CSF was highly expressed in the villus, and expression levels were low in RSA tissues compared to normal tissues. Down-regulation of G-CSF in the trophoblast cell line (HTR-8/SVneo) by siRNA was associated with a decrease in cell activities. Trophoblast-derived exosomes inhibited the activation of the macrophage cell line (RAW264.7), whereas G-CSF free exosomes had no effects on macrophage activation. Intraperitoneal administration of G-CSF improved pregnancy outcomes in RSA mice and increased the amounts of G-CSF at the maternal-foetal interface.
G-CSF levels were downregulated in villi of RSA patients. The absence of G-CSF impaired the proliferation as well as migration capacities of trophoblasts, and weakened the suppression of trophoblasts against macrophages. This implies that suppressed G-CSF levels may be a key factor in RSA occurrence. G-CSF decreased the rate of abortion in RSA mice, thus, it could be a treatment option for RSA patients.
约半数复发性自然流产（RSA）患者的病因尚未明确。粒细胞集落刺激因子（G-CSF）对妊娠具有保护作用，其缺乏可能导致妊娠失败。然而，G-CSF活性的影响及机制尚未完全阐明。因此，我们旨在评估G-CSF缺失是否通过影响母胎界面的细胞通讯诱导RSA。
从参与者获取绒毛和蜕膜组织，通过qRT-PCR、蛋白质免疫印迹和免疫组织化学测定G-CSF的表达水平。用小干扰RNA（siRNA）下调滋养层细胞中G-CSF的水平。从滋养层细胞中提取外泌体并与巨噬细胞共培养。通过qRT-PCR和蛋白质免疫印迹测定关键基因的分子表达水平。通过Transwell和CCK8试验评估细胞的迁移和增殖。对RSA小鼠模型腹腔注射G-CSF，以评估妊娠结局以及母胎界面G-CSF及其受体的表达谱。
与蜕膜相比，G-CSF在绒毛中高表达，与正常组织相比，RSA组织中的表达水平较低。siRNA下调滋养层细胞系（HTR-8/SVneo）中G-CSF的表达与细胞活性降低有关。滋养层来源的外泌体抑制巨噬细胞系（RAW264.7）的活化，而不含G-CSF的外泌体对巨噬细胞活化无影响。腹腔注射G-CSF可改善RSA小鼠的妊娠结局，并增加母胎界面G-CSF的含量。
RSA患者绒毛中G-CSF水平下调。G-CSF的缺失损害了滋养层细胞的增殖和迁移能力，并削弱了滋养层细胞对巨噬细胞的抑制作用。这表明G-CSF水平受抑制可能是RSA发生的关键因素。G-CSF降低了RSA小鼠的流产率，因此，它可能是RSA患者的一种治疗选择。
