H9胚胎干细胞源细胞外囊泡促进子宫内膜损伤修复的研究

H9 embryonic stem cell-derived extracellular vesicles promote endometrial repair

目的:探究H9人胚胎干细胞(H9-hESCs)细胞外囊泡(EVs)对子宫内膜损伤修复的影响。方法:从H9-hESCs培养上清中提取EVs并鉴定。建立小鼠子宫内膜损伤模型,将其双侧子宫分为EVs实验组和PBS对照组,分别进行EVs和PBS注射,HE染色观察子宫内膜组织病理学变化、免疫组化法分析增殖细胞核抗原(PCNA)表达情况。通过EdU法、Western blot法评估EVs对人子宫内膜基质细胞(hEndoSCs)增殖的影响。结果:H9-hESCs-EVs是粒径为(144.7±2.1)nm并且具有膜结构的纳米小体,表达CD63和TSG101特征蛋白。与PBS组相比,EVs实验组子宫内膜组织形态完整、厚度及腺体数量增加,EVs实验组PCNA表达的平均光密度较PBS组显著升高(P<0.05)。EdU、Western blot检测结果显示H9-hESCs-EVs促进hEndoSCs增殖,且与EVs呈现蛋白浓度正相关(P<0.05)。结论:H9-hESCs-EVs通过提高子宫内膜基质细胞增殖促进子宫内膜损伤修复。

AIM:To investigate the reparative effect of extracellular vesicles(EVs)derived from H9 human embryonic stem cells(H9-hESCs)on endometrial injury.METHODS:EVs were isolated from the culture supernatant of H9-hESCs and characterized.A mouse model of endometrial injury was established,with bilateral uterine divisions into an EVs experimental group and a PBS control group.EVs and PBS were injected respectively.Histological changes in the endometrium were assessed using HE staining,and proliferating cell nuclear antigen(PCNA)expression was analyzed via immunohistochemistry.The impact of EVs on the proliferation of human endometrial stromal cells(hEndoSCs)was evaluated using EdU staining and Western blot.RESULTS:H9-hESCs-EVs exhibited a membrane-structured nanobody with a particle size of(144.7±2.1)nm and expressed characteristic proteins CD63 and TSG101.Compared to the PBS control group,the EVs group showed increased endometrial tissue morphology,thickness,and gland numbers.The average optical density of PCNA expression significantly increased in the EVs group compared to the PBS group(P<0.05).Results from EdU staining and Western blot demonstrated that H9-hESCs-EVs promoted hEndoSC proliferation,with a positive correlation observed between H9-hESCs-EVs and EVs protein concentration(P<0.05).CONCLUSION:H9-hESCs-EVs enhance the repair of endometrial injury by stimulating the proliferation of endometrial stromal cells.