脐带华通氏胶间充质干细胞治疗输卵管性不孕初探

Tubal infertility is the most frequent reason which lead to infertility, Assisted reproductive techniques provide the hope of procreation for these patients. However, 'test tube' babies have high risk of birth defect: culture of embryo in vitro may lead to epigenetic errors and its long-term effect on human is still unkown. Therefore it is no doubt that the most ideal way to treat tubal infertility is to repair the impaired fallopian tube and restore its generative function. Wharton's jelly-derived mesenchymal stem cells (WJMSCs) have became the hot spot of cell therapy because of its merit such as easy acquired 、pluripotency properties etc. But until now, there is no report ahout the application of WJMSCs to treat tubal infertility. According to the previous researches in our group which shown that WJMSCs have tendency to transform into oviduct epithelial cells in vitro culture, we suggest a new exploration that using WJMSCs' character such as pluripotency properties、anti-fibrosis and promoting tissue repair to treat tubeal infertility. Methonds: New Zealand rabbits were used to set up the study models, WJMSCs were used for interfereing in these models. The therapeutic efficacy were assessed in morphology、function and molecular biology through varietal empirical methods such as flow cytometry、laser scanning confocal microscope and RT-PCR etc. Through animal experiment in vivo, the feasibility of WJMSCs in repairing impaired oviduct, both acute and chronic inflammatory injury, was investigated.

输卵管不孕是最常见不孕原因，"试管婴儿"技术为这部分患者提供了生育希望，然而异常妊娠风险增加，胚胎体外培养可能导致遗传修饰错误，远期影响仍然未知。因此修复损伤输卵管，恢复其生殖功能，无疑是治疗输卵管不孕最理想手段。脐带华通氏胶间充质干细胞（WJMSCs）因其易获得、多分化潜能及免疫原性低等优点，已成为干细胞治疗热点。但目前国内外尚未见应用WJMSCs治疗输卵管不孕的报道。本课题组前期研究显示，WJMSCs在体外培养具有向输卵管上皮转化的潜能，并且对损伤输卵管上皮有修复作用。据此提出利用WJMSCs多分化潜能、抗纤维化和促组织修复特性，治疗输卵管不孕的全新理念。研究以新西兰兔建立输卵管急慢性炎症损伤模型，应用WJMSCs经三种途径进行干预，采用流式细胞、激光共聚焦、RT-PCR等技术，在功能学、形态学、分子生物学三个水平对修复效果进行评估，探讨WJMSCs修复输卵管急性及慢性损伤的可行性。

一、项目的背景.输卵管不孕是最常见不孕原因，"试管婴儿"技术为这部分患者提供了生育希望，然而胚胎体外培养可能导致遗传修饰错误，远期影响仍然未知。因此修复损伤输卵管，无疑是治疗输卵管不孕最理想手段。脐带华通氏胶间充质干细胞（WJMSCs）已成为干细胞治疗热点。但目前国内外尚未见应用WJMSCs治疗输卵管不孕的报道。..二、主要研究内容.1.以新西兰兔建立输卵管慢性炎症损伤模型，应用WJMSCs进行局部及全身干预治疗，在功能学、形态学、分子生物学水平对修复效果进行评估，探讨WJMSCs修复输卵管急性及慢性损伤的可行性。.2.脐带华通氏胶间充质干细胞外泌体（hUC-MSCs-Ex）对慢性输卵管炎进行治疗，探讨“无细胞移植”干细胞治疗的效果。..三、重要结果.1.功能学方面：.1)分泌功能：慢性输卵管炎的OVGP分泌明显较正常输卵管低。WJMSCs治疗后OVGP的分泌得到不同程度的提高；.2)生育功能：WJMSCs治疗新西兰兔慢性输卵管炎后可基本恢复输卵管的生育功能。.2.形态学方面：WJMSCs治疗后，慢性炎症输卵管粘膜上皮的结构光镜下基本恢复；在电镜下可见WJMSCs治疗能改善或基本恢复输卵管的超微结构；WJMSCs可阻止慢性炎症输卵管的粘膜下胶原纤维增生，同时能有效抑制炎症反应；.3.WJMSCs治疗途径：在同等治疗剂量下，单纯阴道灌注组效果优于静脉注射+阴道灌注组；.4.“无细胞移植”干细胞治疗的探讨，利用脐带间充质干细胞外泌体（hUC-MSCs-Ex）进行治疗，一定程度上能恢复慢性输卵管炎SD雌性大鼠的生殖能力。..四、关键数据.1.培养人才：博士研究生4名，硕士研究生1名；.2.发表论文：发表学术论文5篇，其中3篇SCI收录。研究结果将会后续发表..五、科学意义.首次将华通胶间充质干细胞用于治疗输卵管性不孕，提出一种新的修复输卵管损伤的方法，为进一步临床试验提供依据。首次应用hUC-MSCs-Ex进行“无细胞移植”干细胞治疗的探讨。