背景:作为一种双重功能代谢产物,琥珀酸已经出现在细胞功能中,并在将线粒体功能与其他细胞功能联系起来中起着关键的信号作用。胞质中的琥珀酸积累通常与微环境中的缺氧和免疫细胞活化有关。释放到微环境中的细胞外琥珀酸盐被认为是一种炎症警报,可以通过其膜受体SUCNR1来感知,从而增强促炎反应并类似于经典的激素和细胞因子。琥珀酸盐在炎症性疾病的发生发展中起重要作用。琥珀酸盐是否促进子宫内膜异位症(EMs)的进展,以慢性炎症和腹膜粘连为特征,值得探索。
目的:我们在体外和体内模拟异位环境,以评估琥珀酸在子宫内膜异位症中的主要来源和潜在作用。我们评估了琥珀酸盐对腹膜腔内巨噬细胞和腹膜间皮细胞的分子和功能影响。本研究进一步探讨了琥珀酸盐/SUCNR1信号传导对异位子宫内膜基质细胞(ESC)的影响。
方法:在本研究中,我们使用靶向有机酸代谢组学分析和体外试验来评估琥珀酸在EMs患者腹膜液中的潜在积累.我们检查了它与疾病严重程度的相关性,视觉模拟量表,和子宫内膜异位症生育指数。流式细胞术,酶联免疫吸附测定,westernblot检测,实时定量PCR,和其他分子生物学技术被用来探索潜在的机制。
结果:通过模仿异位环境,我们构建了一个体外共培养系统,发现M1极化的巨噬细胞和腹膜间皮细胞系(HMrSV5)主要释放琥珀酸进入其微环境并激活琥珀酸受体(SUCNR1)信号,进一步使巨噬细胞极化并显着增强ESCs的侵袭性存活,和腹膜的粘连。我们使用子宫内膜异位症的异种移植小鼠模型进一步研究了体内细胞外琥珀酸的病理作用。
结论:琥珀酸-SUCNR1信号传导促进了炎症细胞的产生,并在EMs进展和腹膜粘连中起着至关重要的作用。我们对琥珀酸积累和功能的分子机制的研究将有助于阐明EMs疼痛和不孕症的表型奥秘。
As a dual-function metabolite, succinate has emerged in cell function and plays a key signaling role in linking mitochondrial function to other cellular functions. Succinate accumulation in the cytoplasm is commonly associated with hypoxia in the microenvironment and immune cell activation. Extracellular succinate released into the microenvironment is considered an inflammatory alarm that can be sensed by its membrane receptor
SUCNR1, which boosts proinflammatory responses and acts akin to classical hormones and cytokines. Succinate plays an important role in the development of inflammatory diseases. Whether succinate facilitates the progression of endometriosis (EMs), characterized by chronic inflammation and peritoneal adhesion, is worth exploring.
We mimicked the ectopic milieu in vitro and in vivo to evaluate the main source and potential role of succinate in endometriosis. We assessed the molecular and functional effects of succinate on macrophages and peritoneal mesothelial cells in peritoneal cavity. The effect of succinate/
SUCNR1 signaling on ectopic endometrial stromal cells (ESCs) was further explored in this study.
In this study, we used targeted organic acid metabolomics analysis and in vitro assays to assess the potential accumulation of succinate in the peritoneal fluid of EMs patients. We examined its correlation with disease severity, Visual Analogue Scale, and the Endometriosis Fertility Index. Flow cytometry, enzyme linked immunosorbent assay, western blot assay, quantitative real-time PCR, and other molecular biology techniques were used to explore the potential mechanisms.
By mimicking the ectopic milieu, we constructed an in vitro co-culture system and found that M1 polarized macrophages and that the peritoneal mesothelial cell line (HMrSV5) mainly released succinate into their microenvironment and activated the succinate receptor (
SUCNR1) signal, which further polarized the macrophages and significantly enhanced the invasive survival of ESCs, and the adhesion to the peritoneum. We further investigated the pathological effects of extracellular succinate in vivo using a xenograft mouse models of endometriosis.
Succinate-
SUCNR1 signaling facilitates the creation of inflammatory cells and plays a vital role in EMs progression and peritoneal adhesion. Our work on the molecular mechanisms underlying succinate accumulation and function will help elucidate the phenotypic mysteries of pain and infertility in EMs. Video Abstract.