子宫血管旁多能细胞分泌CYR61促进子宫内膜损伤后血管新生及其机制

Severe scarring of the endometrium causes menstrual abnormalities, infertility and miscarriage. Asherman’s symptom is a difficult and complicated disease. Endometrial vascular regeneration remains the basis of endometrial tissue remodeling and endometrial repair after damage. However, the mechanism is largely unknown. Our primary results showed that endometrial pericytes were multi-differentional cells with high expression of ER-α and angiogenic CYR61, to support the spouting of vascular endothelial cells in vitro. As a leading role in angiogenesis, it is also reported that pericytes spromote proliferation and migration of endothelial cells, increase angiogenesis homeostasis. Therefore, we speculate that perivascular stem cells secrete CYR61 under the stimulation of E2 and hypoxia, binding cell surface receptors integrin αvβ3 and α6β1, promote VEGFC secretion and migration of endothelial cells and increase junction proteins expression, which will support the development and stabilization of new blood vessel. To confirm this hypothesis, a series of experiments, including flow cytometric sorting, CYR61 secretion, integrin antibody blocking, blood vessel permeability testing and cells transplantation will be carried out to clarify the role and mechanism of pericytes in the endometrial angiogenesis after endometrial injury. The present study will provide a new path for endometrial injury interventions.

子宫内膜重度疤痕引起月经异常、不孕和流产，是妇产科疑难疾病。子宫内膜血管新生是子宫内膜重构的基础及损伤后子宫内膜修复的关键，但其机制不明。我们前期的实验结果发现，子宫血管旁存在一群多能细胞，表达间充质干细胞标志物，具有多潜能性，高表达ERα和血管生成因子CYR61，支持血管内皮细胞的体外环化。文献报道，血管旁细胞促进血管内皮细胞增殖、迁移，增加新生血管稳态。据此我们推测：子宫血管旁多能细胞，在E2和低氧刺激下旁分泌CYR61，与血管内皮细胞表面整合素受体αvβ3、α6β1结合，促进血管内皮细胞VEGFC分泌和细胞迁移，增加血管内皮细胞间连接蛋白表达，新生和稳定子宫内膜血管。为验证上述推测，我们通过流式细胞分选术、E2和低氧条件诱导CYR61分泌、整合素抗体封闭、血管通透性检测以及体内移植等实验，研究子宫血管旁多能细胞在子宫内膜损伤后血管新生中的作用和机制，为子宫内膜损伤干预提供新路径。

子宫内膜重度瘢痕引起月经异常、不孕和流产，是妇产科疑难疾病。子宫内膜血管新生是子宫内膜重构的基础及损伤后子宫内膜修复的关键，但其机制不明。我们前期的实验结果发现，子宫血管旁存在一群多能细胞，表达间充质干细胞标志物，具有多潜能性，高表达ERα和血管生成因子CYR61，支持血管内皮细胞的体外环化。文献报道，血管旁细胞促进血管内皮细胞增殖、迁移，增加新生血管稳态。本研究期望分析子宫血管旁多能细胞特征及其在子宫内膜损伤后血管新生中的作用。研究结果发现瘢痕化子宫患者较正常育龄期女性子宫内膜血流参数明显下降，且血管旁多能细胞分布也减少。采用流式细胞术从正常人子宫内膜中分选出血管旁多能细胞（CD45-CD56-CD144-CD34-CD146+），质谱分析结果显示其中CYR61蛋白与血管旁多能细胞促进血管新生密切相关。体外实验结果发现，子宫血管旁多能细胞通过旁分泌 CYR61 促进子宫内膜血管新生，并且CYR61 在子宫内膜血管新生中具有重要作用。此外，探讨高表达CYR61的血管旁多能细胞作为种子细胞联合胶原支架移植修复损伤子宫内膜的可行性，结果证实高表达CYR61的血管旁多能细胞促进模型大鼠子宫损伤后的血管新生，修复子宫内膜的容受性、提高胚胎的种植率，为临床治疗重度子宫内膜疤痕提供新的思路。