蜕膜巨噬细胞通过外泌体调控滋养细胞生物学行为参与不明原因复发性流产

Decidual macrophages modulate trophoblast cells by transferring exosomes in unexplained recurrent spontaneous abortion

目的:研究蜕膜巨噬细胞分泌的外泌体是否调控滋养细胞生物学行为,参与不明原因复发性流产(URSA)的发生。方法:收集正常早孕人工流产妇女和URSA患者的蜕膜组织,采用免疫磁珠法分选巨噬细胞。分离蜕膜巨噬细胞外泌体,透射电镜观察其形态,Western blot检测外泌体标记蛋白CD63的表达。荧光显微镜检测外泌体能否被滋养细胞摄取。将正常蜕膜巨噬细胞外泌体和URSA患者蜕膜巨噬细胞外泌体分别与滋养细胞HTR8/Snveo共培养,采用MTS法检测共培养后1 d、2 d和3 d滋养细胞的细胞活力;Transwell迁移实验检测共培养后滋养细胞的迁移能力。结果:透射电镜显示,外泌体直径40~80 nm,近似圆形。Western blot结果表明蜕膜巨噬细胞外泌体高表达CD63。荧光显微镜结果显示,外泌体可被滋养细胞内吞。MTS实验结果显示,与正常组相比,URSA组蜕膜巨噬细胞外泌体在共培养后第2和第3天均抑制滋养细胞的活力(P<0.05)。Transwell迁移实验结果表明,与对照组相比,URSA组蜕膜巨噬细胞外泌体明显抑制滋养细胞的迁移能力(P<0.01)。结论:蜕膜巨噬细胞可能通过外泌体调控滋养细胞生物学行为,参与母胎免疫调节,与URSA发病相关。

AIM: To investigate whether decidual macrophages have ability to regulate trophoblast cells by secreting exosomes in unexplained recurrent spontaneous abortion (URSA). METHODS: The decidual macrophages were isolated from deciduas of URSA and normal early pregnant induced abortion women. Transmission electron microscopy revealed the morphology and size of the macrophages-derived exosomes. Besides, the characterization of the exosomes was confirmed by the presence of known exosomal marker CD63 by Western blot. The HTR8/Snveo cells were treated with exosomes extracted from the decidual macrophages of URSA or normal early pregnanty induced abortion women. Confocal microscopy, MTS assay and Transwell assay were used to explore the effect of URSA macrophages-derived exosomes on viabi- lity and migration ability of trophoblasts cells. RESULTS: Electron microscopy showed exosomes secreted by macrophages displayed a round-shaped appearance, ranging 40~80 nm in diameter. The results of Western blot indicated that exosomes exacted from the macrophages expressed CD63, a member of tetraspanin family protein. Confocal microscopy revealed the exosomes were taken up by the HTR8/Snveo cells. MTS assay indicated the viability of HTR8/Snveo cells was declined when incubated with the URSA macrophages-derived exosomes ( P <0.05). In URSA macrophages-derived exosomes group, the migration ability of the HTR8/Snveo cells was decreased significantly as compared with normal early pregnant induced abortion macrophages-derived exosome ( P <0.05). CONCLUSION: Decidual macrophages of URSA inhibits the viability and migratory properties of trophoblast cells by exosomes, which participates in regulating maternal-fetal immune in URSA.