背景:子宫内膜异位症患者患有慢性盆腔疼痛和不孕症,以及缺乏持续阻止疾病进展的药物治疗。子宫内膜异位症患者与子宫内膜的差异未受影响的控制是有据可查的。具体来说,脱落的子宫内膜组织(通过逆行月经传递到盆腔)显示,一部分基质细胞表现出促炎,促纤维化,与对照组相比,子宫内膜异位症患者的促衰老样表型增强。此外,来自子宫内膜异位症患者的培养活检来源的子宫内膜基质细胞表现出受损的蜕膜化,人类胚胎植入和妊娠所需的明确分化过程。槲皮素,一种抗衰老剂,显示肺纤维化的治疗潜力,一种归因于衰老肺成纤维细胞的疾病。在子宫内膜异位症的啮齿动物模型中,槲皮素显示出希望,槲皮素可促进体外蜕膜化。然而,确切的机制还不完全清楚。因此,我们研究了槲皮素对子宫内膜异位症患者和未受影响的对照组月经流出物来源的子宫内膜基质细胞的影响,以确定槲皮素对子宫内膜基质细胞影响的信号通路。
方法:从未受影响的对照组和子宫内膜异位症患者收集并培养月经流出物来源的子宫内膜基质细胞,在基础或标准蜕膜化条件下用槲皮素(25µM)处理低传代细胞.通过测量IGFBP1和PRL的产生来分析决策响应。此外,测定槲皮素对细胞内cAMP水平和细胞氧化应激反应的影响。磷酸激酶阵列,西方印迹,和流式细胞术方法来确定槲皮素对各种信号通路的影响以及槲皮素的潜在机制作用。
结果:槲皮素显著促进对照和子宫内膜异位症-子宫内膜基质细胞的蜕膜化。槲皮素显著降低AKT和ERK1/2通路中多个信号分子的磷酸化,同时增强p53的磷酸化和总p53水平。此外,p53抑制阻断蜕膜化,而p53激活促进蜕膜化。最后,我们提供证据表明槲皮素可增加具有衰老样表型的子宫内膜基质细胞的凋亡。
结论:这些数据提供了槲皮素对子宫内膜基质细胞的作用机制的见解,并保证了未来的临床试验,以测试槲皮素和其他促性腺激素治疗子宫内膜异位症。
BACKGROUND: Patients with endometriosis suffer with chronic pelvic pain and infertility, and from the lack of pharmacologic therapies that consistently halt disease progression. Differences in the endometrium of patients with endometriosis vs. unaffected controls are well-documented. Specifically, shed endometrial tissues (delivered to the pelvic cavity via retrograde menstruation) reveal that a subset of stromal cells exhibiting pro-inflammatory, pro-fibrotic, and pro-senescence-like phenotypes is enhanced in endometriosis patients compared to controls. Additionally, cultured biopsy-derived endometrial stromal cells from endometriosis patients exhibit impaired decidualization, a defined differentiation process required for human embryo implantation and pregnancy. Quercetin, a senolytic agent, shows therapeutic potential for pulmonary fibrosis, a disorder attributed to senescent pulmonary fibroblasts. In rodent models of endometriosis, quercetin shows promise, and quercetin improves decidualization in vitro. However, the exact mechanisms are not completely understood. Therefore, we investigated the effects of quercetin on menstrual effluent-derived endometrial stromal cells from endometriosis patients and unaffected controls to define the signaling pathways underlying quercetin\'s effects on endometrial stromal cells.
METHODS: Menstrual effluent-derived endometrial stromal cells were collected and cultured from unaffected controls and endometriosis patients and then, low passage cells were treated with quercetin (25 µM) under basal or standard decidualization conditions. Decidualization responses were analyzed by measuring the production of IGFBP1 and PRL. Also, the effects of quercetin on intracellular cAMP levels and cellular oxidative stress responses were measured. Phosphokinase arrays, western blotting, and flow cytometry methods were performed to define the effects of quercetin on various signaling pathways and the potential mechanistic roles of quercetin.
RESULTS: Quercetin significantly promotes decidualization of control- and endometriosis-endometrial stromal cells. Quercetin substantially reduces the phosphorylation of multiple signaling molecules in the AKT and ERK1/2 pathways, while enhancing the phosphorylation of p53 and total p53 levels. Furthermore, p53 inhibition blocks decidualization while p53 activation promotes decidualization. Finally, we provide evidence that quercetin increases apoptosis of endometrial stromal cells with a senescent-like phenotype.
CONCLUSIONS: These data provide insight into the mechanisms of action of quercetin on endometrial stromal cells and warrant future clinical trials to test quercetin and other senolytics for treating endometriosis.