背景:临床上,激素替代疗法(HRT)是原发性卵巢功能不全(POI)的主要治疗手段。然而,HRT可能会增加患乳腺癌和心血管疾病的风险。人脐带间充质干细胞(hUC-MSC)来源的外泌体通过抑制炎症反应逐渐被应用于多种疾病的治疗,免疫调节,和组织修复功能。然而,hUC-MSC外泌体在POI中的应用和研究仍然有限。
方法:这里,我们首先构建了四种大鼠动物模型:POI-C模型(“环磷酰胺诱导的”腹腔注射POI模型),POI-B模型(“白消安诱导”POI模型),POI-U模型(超声引导下的“环磷酰胺诱导”POI模型),和MS模型(“母体分离模型”)。第二,我们比较了体重,卵巢指数,status,大鼠恶情量表,并发症,不同POI大鼠模型的死亡率。最后,经腹超声引导注射hUC-MSC外泌体,并评价其对POI动物模型的治疗效果,包括激素水平的变化,发情周期,卵巢细胞凋亡水平,和生育能力。此外,我们进行RNA-seq以探索hUC-MSC外泌体功能的可能机制。
结果:与POI-C相比,POI-B,和MS动物模型,POI-U模型显示重量波动较小,较低的卵巢指数,并发症少,较低的死亡率,和更高的模型成功率。第二,我们成功鉴定了hUC-MSCs及其外泌体,并进行超声引导下卵巢内注射hUC-MSCs外泌体。最后,我们证实超声引导的外泌体注射(称为POI-e)有效地改善了卵巢激素水平,发情周期,卵巢功能,和生育能力。机械上,hUC-MSCs可能通过调节卵巢免疫和代谢功能发挥治疗作用。
结论:在我们的研究中,我们创新性地构建了超声引导下卵巢药物注射方法,构建POI-U动物模型和hUC-MSC外泌体注射.并且我们证实了hUC-MSC外泌体对POI-U动物模型的治疗效果。我们的研究将为POI的新型动物模型提供更好的选择,并为POI患者的hUC-MSCs外泌体治疗提供一定的指导。
BACKGROUND: Clinically, hormone replacement therapy (HRT) is the main treatment for primary ovarian insufficiency (POI). However, HRT may increase the risk of both breast cancer and cardiovascular disease. Exosomes derived from human umbilical cord mesenchymal stem cell (hUC-MSC) have been gradually applied to the therapy of a variety of diseases through inflammation inhibition, immune regulation, and tissue repair functions. However, the application and study of hUC-MSC exosomes in POI remain limited.
METHODS: Here, we first constructed four rat animal models: the POI-C model (the \"cyclophosphamide-induced\" POI model via intraperitoneal injection), the POI-B model (the \"busulfan-induced\" POI model), the POI-U model (the \"cyclophosphamide-induced\" POI model under ultrasonic guidance), and MS model (the \"maternal separation model\"). Second, we compared the body weight, ovarian index, status, Rat Grimace Scale, complications, and mortality rate of different POI rat models. Finally, a transabdominal ultrasound-guided injection of hUC-MSC exosomes was performed, and its therapeuticy effects on the POI animal models were evaluated, including changes in hormone levels, oestrous cycles, ovarian apoptosis levels, and fertility. In addition, we performed RNA-seq to explore the possible mechanism of hUC-MSC exosomes function.
RESULTS: Compared with the POI-C, POI-B, and MS animal models, the POI-U model showed less fluctuation in weight, a lower ovarian index, fewer complications, a lower mortality rate, and a higher model success rate. Second, we successfully identified hUC-MSCs and their exosomes, and performed ultrasound-guided intraovarian hUC-MSCs exosomes injection. Finally, we confirmed that the ultrasound-guided exosome injection (termed POI-e) effectively improved ovarian hormone levels, the oestrous cycle, ovarian function, and fertility. Mechanically, hUC-MSCs may play a therapeutic role by regulating ovarian immune and metabolic functions.
CONCLUSIONS: In our study, we innovatively constructed an ultrasound-guided ovarian drug injection method to construct POI-U animal models and hUC-MSC exosomes injection. And we confirmed the therapeutic efficacy of hUC-MSC exosomes on the POI-U animal models. Our study will offer a better choice for new animal models of POI in the future and provides certain guidance for the hUC-MSCs exosome therapy in POI patients.