细胞外囊泡对子宫内膜异位症相关性不孕的作用及其机制  

The Role of Extracellular Vesicles in Endometriosis-related Infertility and Its Mechanism

作　　者：戴昂 张佐 李天鸽 裴钰琳 陈琦[1] DAI Ang;ZHANG Zuo;LI Tian-ge;PEI Yu-lin;CHEN Qi(Department of Obstetrics and Gynecology,the Second Affiliated Hospital of Nanchang University,Nanchang 330006,China)

机构地区：[1]南昌大学第二附属医院妇产科,南昌330006

出　　处：《南昌大学学报（医学版）》2024年第5期57-63,共7页Journal of Nanchang University：Medical Sciences

基　　金：国家自然科学基金资助项目(82260507);江西省卫健委科技计划项目(202310495)。

摘　　要：目的探讨细胞外囊泡(extracellular vesicles,EVs)在子宫内膜异位症(endometriosis,EMs)相关性不孕中的作用及其机制。方法利用Exo-spinTM技术从10例健康育龄女性(H-EVs组)和10例EMs相关性不孕患者(E-EVs组)阴道黏液中分离EVs。以磷酸盐缓冲盐水(PBS)为对照(PBS组,n=10),将人类精子分别与H-EVs、E-EVs和PBS共培养,利用计算机辅助精液分析和伊红染色评估3组精子活力。以10只正常雌性小鼠为空白对照(NC组),将含有60μL PBS、60μL H-EVs溶液、60μL E-EVs溶液的医用可吸收明胶海绵分别送入模型组(M组,n=10)、H-EVs阴道移植组(H-EVs-VT组,n=10)和E-EVs阴道移植组(E-EVs-VT组,n=10)雌鼠阴道,持续1个月;再将雌雄性小鼠同笼配对(雌雄比例4∶1),并在妊娠第9天收集雌鼠子宫、卵巢组织,通过组织病理学、流式细胞术、实时定量反转录PCR及蛋白质印迹法比较4组雌鼠的生育力。结果人EVs形态呈直径为50~200 nm的圆形或椭圆形结构,E-EVs可降低精子运动能力及存活率,而H-EVs能增加精子运动能力及存活率(P<0.05)。动物实验显示:与NC组比较,E-EVs可诱导促炎因子TGFβ1、IL-6、IL-17A和转录因子ROR-γt的释放导致Th17/Treg失衡,降低EMs小鼠的妊娠率及胚胎着床数目(P<0.05);而H-EVs可增加抗炎因子IL-10和转录因子Foxp3的表达从而缓解Th17/Treg失衡,增加EMs小鼠的妊娠率及胚胎着床数目(P<0.05)。结论阴道黏液EVs可调控精子活力,并介导EMs相关的炎症因子释放来调节Th17/Treg平衡,从而影响生育能力,这为EMs相关性不孕的机制提供新的解释。Objective To investigate the role of extracellular vesicles(EVs)in endometriosis(EMs)-associated infertility and its mechanism.Methods Exo spinTM technology was used to isolate EVs from vaginal mucus of 10 healthy women of childbearing age(H-EVs group)and 10 patients with EMs-associated infertility(E-EVs group).With phosphate buffered saline as the control group(PBS group,n=10),human sperms were co-cultured with H-EVs,E-EVs and PBS,respectively,and sperm motility of the 3 groups was evaluated using computer-assisted semen analysis and eosin staining.To construct an EMs mouse model,30 EMs mice were randomly divided into the model group(M group),the vaginal transplantation of H-EVs group(H-EVs-VT group),and the E-EVs vaginal transplantation group(E-EVs-VT group),with 10 mice in each group;medical absorbable gelatin sponges containing 60μL of PBS,60μL of H-EVs solution,and 60μL of E-EVs solution were delivered into the vagina of female mice in the 3 groups,respectively;these solutions were injected for 1 month;the male and female mice were then paired in the same cage(4∶1 ratio of male to female);the uterus and ovary tissues of the female mice were collected on the 9th day of gestation,and the fertility of the female mice in the four groups was compared by histopathology,flow cytometry,real-time quantitative reverse transcription PCR and protein blotting.Results EVs in H-EVs group and E-EVs group exhibited circular or elliptical structures with a diameter of 50~200 nm.E-EVs reduced sperm motility and survival rate,while H-EVs increased sperm motility(P<0.05).Animal experiments shown that compared with the NC group,E-EVs induced the release of pro-inflammatory factors TGFβ1,IL-6,IL-17A and transcription factor ROR-γt,resulting in the Th17/Treg imbalance and decreasing the pregnancy rate and the number of embryos implanted in EMs mice(P<0.05),whereas H-EVs increased the expression of anti-inflammatory factor IL-10 and transcription factor Foxp3,thereby alleviating the Th17/Treg imbalance and increasing the preg

关 键 词：子宫内膜异位症 不孕 细胞外囊泡 人类精子 TH17/TREG 动物 实验 EMs小鼠 

分 类 号：R711.6[医药卫生—妇产科学] R-332[医药卫生—临床医学]