背景:当异常发生在增殖中时,滋养细胞的分化和凋亡,胎盘滋养层的侵袭能力减弱,易引发各种妊娠疾病的发生,如反复自然流产(RSA)。临床上,黄芪和党参多糖(APS和CPPS)用于治疗原因不明的复发性自然流产(URSA)。因此,目的探讨APS和CPPS在胎盘滋养细胞生物学行为中的作用。
方法:用APS和CPPS治疗滋养细胞,并转染miR-92a-1-5p模拟物和CCR7质粒,探讨APS和CPPS的作用。通过CCK-8和流式细胞术测定细胞活力和凋亡,分别。通过qRT-PCR和westernblot检测miRNA/mRNA和蛋白质水平,分别。通过TargetScan和双荧光素酶报告基因测定分析miR-92a-1-5p与CCR7之间的相互作用。通过Transwell和伤口愈合试验评估侵袭和迁移率,分别。
结果:APS联合CPPS增强了活力,Bcl-2表达,以及迁移和入侵能力,在抑制细胞凋亡的同时,和Bax的表达,滋养细胞中的Bim和miR-92a-1-5p。然而,miR-92a-1-5p模拟物在滋养细胞中产生反向调制,部分逆转了APS和CPPS的影响。此外,过表达CCR7,miR-92a-1-5p的靶标,部分抵消miR-92a-1-5p模拟物在滋养细胞中的作用。
结论:黄芪联合党参多糖通过miR-92a-1-5p/CCR7轴调节滋养细胞的生物学行为。我们研究的调节轴将有助于将来URSA的治疗。
BACKGROUND: When the abnormality occurs in proliferation, differentiation and apoptosis of trophoblasts, the invasion ability of placental trophoblast is weakened, which is prone to trigger the occurrence of various pregnancy diseases such as repeated spontaneous abortion (RSA). Clinically, Astragalus and Codonopsis pilosula polysaccharides (APS and CPPS) are used for the treatment of unexplained recurrent spontaneous abortion (URSA). Therefore, we aimed to probe into the roles of APS and CPPS in biological behaviors of placental trophoblasts.
METHODS: The trophoblasts were treated with APS and CPPS, and transfected with miR-92a-1-5p mimic and CCR7 plasmid to explore the roles of APS and CPPS. Cell viability and apoptosis were determined by CCK-8 and flow cytometry, respectively. The levels of miRNA/mRNA and protein were measured by qRT-PCR and western blot, respectively. The interaction between miR-92a-1-5p and CCR7 was analyzed by TargetScan and dual-luciferase reporter assay. Invasion and migration rates were assessed by Transwell and wound healing assays, respectively.
RESULTS: APS combined with CPPS enhanced viability, Bcl-2 expression, and migration and invasion capabilities, while suppressing apoptosis, and expressions of Bax, Bim and miR-92a-1-5p in trophoblasts. Nevertheless, miR-92a-1-5p mimic produced the inverse modulations in trophoblasts, and partially reversed the effects of APS and CPPS. Furthermore, overexpression of CCR7, the target of miR-92a-1-5p, partially offset the effect of miR-92a-1-5p mimic in trophoblasts.
CONCLUSIONS: Astragalus combined with Codonopsis pilosula polysaccharides modulates the biological behaviors of trophoblasts via miR-92a-1-5p/CCR7 axis. The regulatory axis we studied will be helpful for the treatment of URSA in the future.