Shahgaldi Shahab, Rezaei Kahmini Fatemeh, Moazzeni Seyed Mohammad
Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Mol Immunol. 2022 Jan;141:246-256. doi: 10.1016/j.molimm.2021.11.010. Epub 2021 Dec 6.
Immunological disorders are one of the main causes of recurrent spontaneous abortions (RSA). A rapidly expanding body of evidence indicates that excessive activation of the complement system is critically involved in the development of miscarriages. In the CBA/J × DBA/2 murine model of recurrent miscarriage, exaggerated and unrestrained complement activation is reported to be the underlying cause of angiogenic factor imbalance and persistent inflammation. We have previously shown that mesenchymal stem cell (MSC) therapy can significantly reduce the abortion rate in abortion-prone mice through regulating the feto-maternal immune response. In the present study, we hypothesized that MSCs might improve the balance of angiogenic factors at the feto-maternal unit of CBA/J × DBA/2 mice by restraining complement activation and deposition. To explore this hypothesis, autologous adipose tissue-derived mesenchymal stem cells (AD-MSCs) were administered intra-peritoneally to abortion-prone mice on the 4.5th day of gestation. Control mice received PBS as vehicle. On day 13.5 of pregnancy, deposition of the complement component C3 and expression levels of Crry, CFD (adipsin), VEGF, PlGF and FLT-1 were measured at the feto-maternal interface by immunohistochemistry and real-time PCR, respectively. Decidual cells were also cultured in RPMI 1640 medium for 48 h and VEGF and sFLT-1 protein levels were quantified in supernatants using enzyme-linked immunosorbent assay (ELISA). Our results indicated that MSC therapy significantly reduced C3 deposition and adipsin transcription in the fetal-maternal interface of abortion-prone mice. Furthermore, administration of MSCs robustly upregulated the mRNA expression levels of Crry, VEGF, PlGF and FLT-1 in the placenta and decidua of CBA/J × DBA/2 mice. Consistently, the in vitro results demonstrated that decidual cells obtained from MSC-treated dams produced increased concentrations of VEGF in culture supernatants, with concomitant decreased levels of sFLT-1 protein. Here, we show for the first time that adoptive transfer of MSCs rectifies the disturbed balance of angiogenic factors observed at the feto-maternal unit of CBA/J × DBA/2 mice, in part at least, through inhibiting excessive complement activation and promoting the production of angiogenic factors. Collectively, these alterations seem to play a pivotal role in reducing the abortion rate and improving the intrauterine condition for the benefit of the fetus.
免疫紊乱是复发性自然流产（RSA）的主要原因之一。越来越多的证据表明，补体系统的过度激活在流产的发生发展中起关键作用。在CBA/J×DBA/2复发性流产小鼠模型中，据报道，过度且不受控制的补体激活是血管生成因子失衡和持续性炎症的根本原因。我们之前已经表明，间充质干细胞（MSC）疗法可以通过调节母胎免疫反应显著降低易流产小鼠的流产率。在本研究中，我们假设MSC可能通过抑制补体激活和沉积来改善CBA/J×DBA/2小鼠母胎单位血管生成因子的平衡。为了探索这一假设，在妊娠第4.5天给易流产小鼠腹腔注射自体脂肪组织来源的间充质干细胞（AD-MSC）。对照小鼠接受PBS作为载体。在妊娠第13.5天，分别通过免疫组织化学和实时PCR在母胎界面测量补体成分C3的沉积以及Crry、CFD（脂肪酶）、VEGF、PlGF和FLT-1的表达水平。蜕膜细胞也在RPMI 1640培养基中培养48小时，并使用酶联免疫吸附测定（ELISA）定量上清液中VEGF和sFLT-1蛋白水平。我们的结果表明，MSC疗法显著减少了易流产小鼠母胎界面的C3沉积和脂肪酶转录。此外，给予MSC强烈上调了CBA/J×DBA/2小鼠胎盘和蜕膜中Crry、VEGF、PlGF和FLT-1的mRNA表达水平。一致地，体外结果表明，从接受MSC治疗的母鼠获得的蜕膜细胞在培养上清液中产生的VEGF浓度增加，同时sFLT-1蛋白水平降低。在这里，我们首次表明，MSC的过继转移至少部分地通过抑制过度的补体激活和促进血管生成因子的产生，纠正了在CBA/J×DBA/2小鼠母胎单位观察到的血管生成因子的紊乱平衡。总的来说，这些改变似乎在降低流产率和改善子宫内环境以利于胎儿方面起着关键作用。
