Osteoprotegerin interacts with syndecan-1 to promote human endometrial stromal decidualization by decreasing Akt phosphorylation
Osteoprotegerin interacts with syndecan-1 to promote human endometrial stromal decidualization by decreasing Akt phosphorylation
复制标题
骨保护素与 syndecan-1 相互作用，通过降低 Akt 磷酸化促进人子宫内膜基质蜕膜化
DOI：
10.1093/humrep/deaa233
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发表时间：
2020-11-01
影响因子：
6.1
通讯作者：
Shi, Guixiu
中科院分区：
文献类型：
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作者：
Jiang, Yufei;Li, Jianing;Shi, Guixiu
研究问题：骨保护素（OPG）是否促进人子宫内膜间质蜕膜化？总结回答：OPG通过与syndecan- 1相互作用降低Akt的磷酸化水平，对子宫内膜间质的蜕膜化起重要作用。已知：OPG（一种细胞因子受体）水平在妊娠妇女的循环中显著升高。然而，OPG在人子宫内膜间质细胞（ESC）蜕膜化中的作用和机制仍不清楚。我们分析了从月经周期规律的妇女中收集的子宫内膜组织样本中OPG的表达（25至35天不等），和从正常早孕或复发性流产（RPL）妇女中采集的蜕膜组织样本她于2018年1月至10月访问了三级护理中心的妇产科。所有受试者在术前均未接受激素治疗至少3个月。共有16名正常早孕妇女和15名RPL妇女被选为本研究的受试者。OPG在蜕膜化的功能进行了探讨，在人子宫内膜基质细胞（HESC）线和原代培养HESCs.PARTICIPANTS/材料，设置，方法：我们收集子宫内膜组织（活检）从受试者在月经周期和蜕膜组织从受试者与正常早孕和RPL在扩张和刮宫的时间。对照组包括随机选择的妇女谁经历了终止一个明显正常的早期妊娠。采用免疫组织化学染色和定量RT-PCR（qRT-PCR）检测子宫内膜OPG的表达。采用免疫荧光染色、western blot和qRT-PCR方法分别检测OPG在永生化人胚胎干细胞系和原代培养人胚胎干细胞增殖和蜕膜化过程中的mRNA和蛋白表达。通过siRNA介导的敲低实验检测OPG在HESC增殖和蜕膜化中的作用。采用流式细胞术和MTS细胞增殖实验进一步研究OPG在HESC增殖中的作用。我们还通过qRT-PCR分析了蜕膜标志物基因表达，以评估OPG缺失对HESC蜕膜化的影响。免疫共沉淀（IP）试验用于确定OPG和Syndecan-1之间的潜在相互作用。Western印迹分析的救援实验进行使用磷脂酰肌醇3-激酶（PI 3 K）信号特异性抑制剂LY 294002被用来调查下游信号通路，通过OPG可以介导的HESC decidualization.Main结果和机会的作用：OPG表达在人类子宫内膜和体外蜕膜化的ESCs。敲除实验显示，OPG缺失使IGF结合蛋白-1（IGFBP- 1）和催乳素（PRL）的表达降低（P
STUDY QUESTION: Does osteoprotegerin (OPG) promote human endometrial stromal decidualization?SUMMARY ANSWER: OPG is essential for human endometrial stromal decidualization through its interaction with syndecan- 1 to decrease Akt phosphorylation.WHAT IS KNOWN ALREADY: OPG (a cytokine receptor) levels are significandy increased in the circulation of pregnant women. However, the role and mechanism of OPG in human endometrial stromal cell (ESC) decidualization remain elusive.STUDY DESIGN, SIZE, DURATION: We analyzed the endometrial expression of OPG in endometrial tissue samples collected from women with regular menstrual cycles (ranging from 25 to 35 days), and decidual tissue samples collected from woman with normal early pregnancy or recurrent pregnancy loss (RPL) who visited the Department of Gynecology and Obstetrics at a tertiary care center from January to October 2018. None of the subjects had hormonal treatment for at least 3 months prior to the procedure. In total, 16 women with normal early pregnancy and 15 with RPL were selected as subjects for this study. The function of OPG in decidualization was explored in a human endometrial stromal cell (HESC) line and primary cultures of HESCs.PARTICIPANTS/MATERIALS, SETTING, METHODS: We collected endometrial tissues (by biopsy) from the subjects during their menstrual cycle and decidual tissues from subjects with a normal early pregnancy and those with RPL at the time of dilation and curettage. The control group comprised randomly selected women who underwent termination of an apparently normal early pregnancy. The endometrial OPG expression was analyzed using immunohistochemical staining and quantitative RT-PCR (qRT-PCR). Immunofluorescence staining and western blot, and qRT-PCR were used to explore the mRNA and protein expression, respectively, of OPG in an immortalized HESC line and in primary cultures of HESC during proliferation and decidualization. siRNA-mediated knockdown experiments were performed to examine the function of OPG in HESC proliferation and decidualization. Flow cytometry and the cell proliferation MTS assay were performed to further examine the role of OPG in HESC proliferation. We also analyzed decidual marker gene expression by qRT-PCR to assess the consequences of OPG loss for HESC decidualization. A co-immunoprecipitation (IP) assay was used to determine the potential interaction between the OPG and Syndecan-1. Western blot analysis of the rescue experiments performed using the phosphatidylinositol 3-kinase (PI3K) signaling-specific inhibitor LY294002 was used to investigate the downstream signaling pathways through which OPG could mediate HESC decidualization.MAIN RESULTS AND THE ROLE OF CHANCE: OPG was expressed in both the human endometrium and in vitro decidualized ESCs. Knockdown experiments revealed that OPG loss impaired the expression of IGF-binding protein-1 (IGFBP- 1) (P < 0.05) and prolactin (PRL) (P