Abstract：
OBJECTIVE: What is the pathological mechanism involved in a thin endometrium, particularly under ischaemic conditions?
CONCLUSIONS: Endometrial dysfunction in patients with thin endometrium primarily results from remodelling in cytoskeletons and cellular junctions of endometrial epithelial cells under ischemic conditions.
BACKGROUND: A healthy endometrium is essential for successful embryo implantation and subsequent pregnancy; ischemic conditions in a thin endometrium compromise fertility outcomes.
METHODS: We recruited 10 patients with thin endometrium and 15 patients with healthy endometrium. Doppler ultrasound and immunohistochemical results confirmed the presence of insufficient endometrial blood perfusion in patients with thin endometrium. Organoids were constructed using healthy endometrial tissue and cultured under oxygen-glucose deprivation (OGD) conditions for 24 h. The morphological, transcriptomic, protein expression, and signaling pathway changes in the OGD organoids were observed. These findings were validated in both thin endometrial tissue and healthy endometrial tissue samples.
METHODS: Endometrial thickness and blood flow were measured during the late follicular phase using transvaginal Doppler ultrasound. Endometrial tissue was obtained via hysteroscopy. Fresh endometrial tissues were used for the generation and culture of human endometrial organoids. Organoids were cultured in an appropriate medium and subjected to OGD to simulate ischemic conditions. Apoptosis and cell death were assessed using Annexin-V/propidium iodide staining. Immunofluorescence analysis, RNA sequencing, western blotting, simple westerns, immunohistochemistry, and electron microscopy were conducted to evaluate cellular and molecular changes.
RESULTS: Patients with thin endometrium showed significantly reduced endometrial thickness and altered blood flow patterns compared to those with healthy endometrium. Immunohistochemical staining revealed fewer CD34-positive blood vessels and glands in the thin endometrium group. Organoids cultured under OGD conditions exhibited significant morphological changes, increased apoptosis, and cell death. RNA-seq identified differentially expressed genes related to cytoskeletal remodeling and stress responses. OGD induced a strong cytoskeletal reorganization, mediated by the RhoA/ROCK signaling pathway. Additionally, electron microscopy indicated compromised epithelial integrity and abnormal cell junctions in thin endometrial tissues. Upregulation of hypoxia markers (HIF-1α and HIF-2α) and activation of the RhoA/ROCK pathway were also observed in thin endometrial tissues, suggesting ischemia and hypoxia as underlying mechanisms.
METHODS: none.
CONCLUSIONS: The study was conducted in an in vitro model, which may not fully replicate the complexity of in vivo conditions.
CONCLUSIONS: This research provides a new three-dimensional in vitro model of thin endometrium, as well as novel insights into the pathophysiological mechanisms of endometrial ischaemia in thin endometrium, offering potential avenues for identifying therapeutic targets for treating fertility issues related to thin endometrium.
BACKGROUND: This study was supported by the National Natural Science Foundation of China (81925013); National Key Research and Development Project of China (2022YFC2702500, 2021YFC2700303, 2021YFC2700601); the Capital Health Research and Development Project (SF2022-1-4092); the National Natural Science Foundation of China (82288102, 81925013, 82225019, 82192873); Special Project on Capital Clinical Diagnosis and Treatment Technology Research and Transformation Application (Z211100002921054); the Frontiers Medical Center, Tianfu Jincheng Laboratory Foundation(TFJC2023010001). The authors declare that no competing interests exist.
CONCLUSIONS: Endometrial dysfunction in patients with thin endometrium primarily results from remodelling in cytoskeletons and cellular junctions of endometrial epithelial cells under ischemic conditions.
BACKGROUND: A healthy endometrium is essential for successful embryo implantation and subsequent pregnancy; ischemic conditions in a thin endometrium compromise fertility outcomes.
METHODS: We recruited 10 patients with thin endometrium and 15 patients with healthy endometrium. Doppler ultrasound and immunohistochemical results confirmed the presence of insufficient endometrial blood perfusion in patients with thin endometrium. Organoids were constructed using healthy endometrial tissue and cultured under oxygen-glucose deprivation (OGD) conditions for 24 h. The morphological, transcriptomic, protein expression, and signaling pathway changes in the OGD organoids were observed. These findings were validated in both thin endometrial tissue and healthy endometrial tissue samples.
METHODS: Endometrial thickness and blood flow were measured during the late follicular phase using transvaginal Doppler ultrasound. Endometrial tissue was obtained via hysteroscopy. Fresh endometrial tissues were used for the generation and culture of human endometrial organoids. Organoids were cultured in an appropriate medium and subjected to OGD to simulate ischemic conditions. Apoptosis and cell death were assessed using Annexin-V/propidium iodide staining. Immunofluorescence analysis, RNA sequencing, western blotting, simple westerns, immunohistochemistry, and electron microscopy were conducted to evaluate cellular and molecular changes.
RESULTS: Patients with thin endometrium showed significantly reduced endometrial thickness and altered blood flow patterns compared to those with healthy endometrium. Immunohistochemical staining revealed fewer CD34-positive blood vessels and glands in the thin endometrium group. Organoids cultured under OGD conditions exhibited significant morphological changes, increased apoptosis, and cell death. RNA-seq identified differentially expressed genes related to cytoskeletal remodeling and stress responses. OGD induced a strong cytoskeletal reorganization, mediated by the RhoA/ROCK signaling pathway. Additionally, electron microscopy indicated compromised epithelial integrity and abnormal cell junctions in thin endometrial tissues. Upregulation of hypoxia markers (HIF-1α and HIF-2α) and activation of the RhoA/ROCK pathway were also observed in thin endometrial tissues, suggesting ischemia and hypoxia as underlying mechanisms.
METHODS: none.
CONCLUSIONS: The study was conducted in an in vitro model, which may not fully replicate the complexity of in vivo conditions.
CONCLUSIONS: This research provides a new three-dimensional in vitro model of thin endometrium, as well as novel insights into the pathophysiological mechanisms of endometrial ischaemia in thin endometrium, offering potential avenues for identifying therapeutic targets for treating fertility issues related to thin endometrium.
BACKGROUND: This study was supported by the National Natural Science Foundation of China (81925013); National Key Research and Development Project of China (2022YFC2702500, 2021YFC2700303, 2021YFC2700601); the Capital Health Research and Development Project (SF2022-1-4092); the National Natural Science Foundation of China (82288102, 81925013, 82225019, 82192873); Special Project on Capital Clinical Diagnosis and Treatment Technology Research and Transformation Application (Z211100002921054); the Frontiers Medical Center, Tianfu Jincheng Laboratory Foundation(TFJC2023010001). The authors declare that no competing interests exist.
摘要：
目的：薄型子宫内膜的病理机制是什么，
结论:子宫内膜薄型患者的子宫内膜功能障碍主要是由于缺血条件下子宫内膜上皮细胞的细胞骨架和细胞连接重塑所致.
背景：健康的子宫内膜对于成功的胚胎植入和随后的妊娠至关重要；薄子宫内膜的缺血状况会损害生育结果。
方法：我们招募了10例子宫内膜薄型患者和15例子宫内膜健康患者。多普勒超声和免疫组织化学结果证实子宫内膜薄型患者存在子宫内膜血液灌注不足。使用健康的子宫内膜组织构建类器官，并在氧糖剥夺（OGD）条件下培养24h。转录组，蛋白质表达，并观察到OGD类器官的信号通路变化。这些发现在薄子宫内膜组织和健康子宫内膜组织样本中都得到了验证。
方法：使用经阴道多普勒超声测量卵泡晚期的子宫内膜厚度和血流量。通过宫腔镜获得子宫内膜组织。新鲜的子宫内膜组织用于人子宫内膜类器官的产生和培养。在适当的培养基中培养类器官并进行OGD以模拟缺血条件。使用膜联蛋白-V/碘化丙啶染色评估细胞凋亡和细胞死亡。免疫荧光分析,RNA测序，西方印迹,简单的西部片,免疫组织化学，和电子显微镜检查以评估细胞和分子的变化。
结果：子宫内膜薄型患者与健康子宫内膜患者相比，子宫内膜厚度明显减少，血流模式改变。免疫组织化学染色显示，薄子宫内膜组中CD34阳性血管和腺体较少。在OGD条件下培养的类器官表现出明显的形态学变化，细胞凋亡增加，细胞死亡。RNA-seq鉴定了与细胞骨架重塑和应激反应相关的差异表达基因。OGD诱导了强烈的细胞骨架重组，由RhoA/ROCK信号通路介导。此外,电子显微镜显示薄子宫内膜组织中上皮完整性受损和细胞连接异常。在薄子宫内膜组织中也观察到缺氧标志物（HIF-1α和HIF-2α）的上调和RhoA/ROCK途径的激活，提示缺血和缺氧是潜在的机制。
方法：无。
结论：该研究是在体外模型中进行的，这可能无法完全复制体内条件的复杂性。
结论：本研究提供了薄子宫内膜的三维体外模型，以及对薄子宫内膜中子宫内膜缺血的病理生理机制的新见解，为确定与薄型子宫内膜相关的生育问题的治疗靶点提供了潜在的途径。
背景：本研究得到了国家自然科学基金（81925013）；国家重点研究开发项目（2022YFC2702500，2021YFC2700303，2021YFC2700601）；首都卫生研究与发展项目（SF2022-1-4092）；国家自然科学基金（8282219102，医学前沿诊断和应用项目，82900022502天府金城实验室基金会(TFJC2023010001)。作者宣称不存在竞争利益。
结论:子宫内膜薄型患者的子宫内膜功能障碍主要是由于缺血条件下子宫内膜上皮细胞的细胞骨架和细胞连接重塑所致.
背景：健康的子宫内膜对于成功的胚胎植入和随后的妊娠至关重要；薄子宫内膜的缺血状况会损害生育结果。
方法：我们招募了10例子宫内膜薄型患者和15例子宫内膜健康患者。多普勒超声和免疫组织化学结果证实子宫内膜薄型患者存在子宫内膜血液灌注不足。使用健康的子宫内膜组织构建类器官，并在氧糖剥夺（OGD）条件下培养24h。转录组，蛋白质表达，并观察到OGD类器官的信号通路变化。这些发现在薄子宫内膜组织和健康子宫内膜组织样本中都得到了验证。
方法：使用经阴道多普勒超声测量卵泡晚期的子宫内膜厚度和血流量。通过宫腔镜获得子宫内膜组织。新鲜的子宫内膜组织用于人子宫内膜类器官的产生和培养。在适当的培养基中培养类器官并进行OGD以模拟缺血条件。使用膜联蛋白-V/碘化丙啶染色评估细胞凋亡和细胞死亡。免疫荧光分析,RNA测序，西方印迹,简单的西部片,免疫组织化学，和电子显微镜检查以评估细胞和分子的变化。
结果：子宫内膜薄型患者与健康子宫内膜患者相比，子宫内膜厚度明显减少，血流模式改变。免疫组织化学染色显示，薄子宫内膜组中CD34阳性血管和腺体较少。在OGD条件下培养的类器官表现出明显的形态学变化，细胞凋亡增加，细胞死亡。RNA-seq鉴定了与细胞骨架重塑和应激反应相关的差异表达基因。OGD诱导了强烈的细胞骨架重组，由RhoA/ROCK信号通路介导。此外,电子显微镜显示薄子宫内膜组织中上皮完整性受损和细胞连接异常。在薄子宫内膜组织中也观察到缺氧标志物（HIF-1α和HIF-2α）的上调和RhoA/ROCK途径的激活，提示缺血和缺氧是潜在的机制。
方法：无。
结论：该研究是在体外模型中进行的，这可能无法完全复制体内条件的复杂性。
结论：本研究提供了薄子宫内膜的三维体外模型，以及对薄子宫内膜中子宫内膜缺血的病理生理机制的新见解，为确定与薄型子宫内膜相关的生育问题的治疗靶点提供了潜在的途径。
背景：本研究得到了国家自然科学基金（81925013）；国家重点研究开发项目（2022YFC2702500，2021YFC2700303，2021YFC2700601）；首都卫生研究与发展项目（SF2022-1-4092）；国家自然科学基金（8282219102，医学前沿诊断和应用项目，82900022502天府金城实验室基金会(TFJC2023010001)。作者宣称不存在竞争利益。
