摘要
目的 探讨骨形态发生蛋白4(bone morphogenetic protein 4,BMP4)在复发性流产(recurrent spontaneous abortion, RSA)患者的人子宫内膜基质细胞(human endometrial stromal cells, hESCs)中的表达及对叉头盒转录因子1(forkhead transcription factors, FOXO1)的调控。方法 收集武汉大学人民医院RSA患者和健康对照(healthy control, HC)患者的蜕膜组织,通过免疫组化和RT-qPCR分析蜕膜中BMP4的表达。利用hESC进行体外细胞实验,通过转染慢病毒敲低BMP4,建立RSA细胞模型,蜕膜化后,通过CCK-8实验、倒置显微镜和Western blot法检测各组细胞的增殖、形态及蜕膜化标志基因的表达。验证BMP4敲低是否影响FOXO1表达,并使用FOXO1抑制剂(AS1842856)进行回复实验,CCK-8实验、倒置显微镜和Western blot法检测分析FOXO1与BMP4的上、下游关系。动物试验中,对孕鼠腹腔注射LPS诱导流产,检测BMP4在小鼠蜕膜中的表达,探究BMP4与小鼠流产的关系。结果 与HC组比较,RSA患者的蜕膜中BMP4表达量明显降低,敲低BMP4后,细胞蜕膜化异常,表现为增殖速度上升,细胞形态转变不明显,且蜕膜化标志基因表达下调。FOXO1为BMP4的下游调控分子,敲低BMP4抑制FOXO1表达,且BMP4可通过FOXO1调控蜕膜化,动物模型中,流产孕鼠蜕膜中BMP4表达量及胎盘重量显著降低。结论 BMP4在RSA患者和流产小鼠蜕膜中的表达下调,细胞实验中,BMP4可通过调控FOXO1的表达促进蜕膜化,BMP4异常低表达可能为RSA患者蜕膜化缺陷的原因之一。
Objective To investigate the expression of Bone morphogenetic protein 4(BMP4)in human endometrial stromal cells(hESCs)of patients with recurrent spontaneous abortion(RSA)and the regulation of Forkhead transcription factor 1(FOXO1)by BMP4.Methods The decidua tissues of RSA patients and Healthy control(HC)patients in People′s Hospital of Wuhan University were collected,and the expression of BMP4 was analyzed by immunohistochemistry and RT-qPCR.hESC was used in vitro cell experiments,lentivirus was transfected to knock down the expression of BMP4 and establish the cell model of RSA.After decidualization,CCK-8 assay,inverted microscope,and Western blot assay were used to detect the cell proliferation,morphology,and expression levels of decidualization marker genes.To verify whether BMP4 knockdown affects FOXO1 expression,a FOXO1 inhibitor(AS1842856)was used to perform a response experiment,and the upstream and downstream relationship between FOXO1 and BMP4 was analyzed by CCK-8 experiment,inverted microscope,and Western blot.In animal experiments,intraperitoneal injection of LPS-induced abortion in pregnant mice,and the expression of BMP4 in decidua were detected to explore the relationship between BMP4 and abortion in mice.Results Compared with the HC group,the expression of BMP4 in the deciduous tissues of RSA patients was significantly decreased.After BMP4 was knocked down,the decidualization was significantly abnormal,which showed that the proliferation rate increased,the cell morphological transformation was not obvious,and the expression of decidualization marker genes was down-regulated.FOXO1 was the downstream regulatory molecule of BMP4,knocking down BMP4 could inhibit the expression of FOXO1,and BMP4 could regulate the decidualization of hESC through FOXO1.In animal models,the expression of BMP4 in decidua and placental weight of aborted pregnant mice were significantly reduced.Conclusion The expression of BMP4 was significantly down-regulated in decidual tissues of RSA patients and aborted mice.In cell experiments,BMP4 can promote decidualization of hESC by regulating the expression of FOXO1.Abnormally low expression of BMP4 may be one of the reasons for decidualization defects in RSA patients.
作者
黄燕洁
代芳芳
陈丽萍
程艳香
HUANG Yanjie;DAI Fangfang;CHEN Liping(Department of Obstetrics and Gynecology,Renmin Hospital of Wuhan University,Hubei 430060,China)
出处
《医学研究杂志》
2024年第10期33-39,共7页
Journal of Medical Research
基金
国家自然科学基金资助项目(面上项目)(82071655)。
