Department of Obstetrics and Gynecology, The University-Town Hospital of Chongqing Medical University, No. 55, Daxuecheng Middle Road, Chongqing, 401331, China.
Cell Commun Signal. 2024 May 31;22(1):301. doi: 10.1186/s12964-024-01656-0.
Intrauterine adhesion (IUA) is one of the most severe causes of infertility in women of childbearing age with injured endometrium secondary to uterine performance. Stem cell therapy is effective in treating damaged endometrium. The current reports mainly focus on the therapeutic effects of stem cells through paracrine or transdifferentiation, respectively. This study investigates whether paracrine or transdifferentiation occurs preferentially in treating IUA.
Human amniotic mesenchymal stem cells (hAMSCs) and transformed human endometrial stromal cells (THESCs) induced by transforming growth factor beta (TGF-β1) were co-cultured in vitro. The mRNA and protein expression levels of Fibronectin (FN), Collagen I, Cytokeratin19 (CK19), E-cadherin (E-cad) and Vimentin were detected by Quantitative real-time polymerase chain reaction (qPCR), Western blotting (WB) and Immunohistochemical staining (IHC). The Sprague-Dawley (SD) rats were used to establish the IUA model. hAMSCs, hAMSCs-conditional medium (hAMSCs-CM), and GFP-labeled hAMSCs were injected into intrauterine, respectively. The fibrotic area of the endometrium was evaluated by Masson staining. The number of endometrium glands was detected by hematoxylin and eosin (H&E). GFP-labeled hAMSCs were traced by immunofluorescence (IF). hAMSCs, combined with PPCNg (hAMSCs/PPCNg), were injected into the vagina, which was compared with intrauterine injection.
qPCR and WB revealed that FN and Collagen I levels in IUA-THESCs decreased significantly after co-culturing with hAMSCs. Moreover, CK19, E-cad, and Vimentin expressions in hAMSCs showed no significant difference after co-culture for 2 days. 6 days after co-culture, CK19, E-cad and Vimentin expressions in hAMSCs were significantly changed. Histological assays showed increased endometrial glands and a remarkable decrease in the fibrotic area in the hAMSCs and hAMSCs-CM groups. However, these changes were not statistically different between the two groups. In vivo, fluorescence imaging revealed that GFP-hAMSCs were localized in the endometrial stroma and gradually underwent apoptosis. The effect of hAMSCs by vaginal injection was comparable to that by intrauterine injection assessed by H&E staining, MASSON staining and IHC.
Our data demonstrated that hAMSCs promoted endometrial repair via paracrine, preferentially than transdifferentiation.
宫腔粘连（IUA）是生育期妇女最严重的不孕原因之一，其子宫内膜因子宫功能受损而发生粘连。干细胞治疗对受损的子宫内膜具有治疗效果。目前的报告主要集中在干细胞通过旁分泌或转分化分别发挥治疗作用。本研究旨在探讨旁分泌或转分化在治疗 IUA 中是否更占优势。
体外共培养人羊膜间充质干细胞（hAMSCs）和转化生长因子β（TGF-β1）诱导的转化人子宫内膜基质细胞（THESCs）。采用实时定量聚合酶链反应（qPCR）、Western blot（WB）和免疫组织化学染色（IHC）检测纤维连接蛋白（FN）、I 型胶原、细胞角蛋白 19（CK19）、E-钙黏蛋白（E-cad）和波形蛋白的 mRNA 和蛋白表达水平。采用 Sprague-Dawley（SD）大鼠建立 IUA 模型。分别将 hAMSCs、hAMSCs 条件培养基（hAMSCs-CM）和 GFP 标记的 hAMSCs 注射到子宫内。通过 Masson 染色评估子宫内膜纤维化面积。通过苏木精和伊红（H&E）染色检测子宫内膜腺体数量。通过免疫荧光（IF）追踪 GFP 标记的 hAMSCs。将 hAMSCs 与 PPCNg（hAMSCs/PPCNg）联合注射到阴道中，与子宫内注射进行比较。
qPCR 和 WB 结果显示，IUA-THESCs 与 hAMSCs 共培养后 FN 和 I 型胶原水平显著降低。此外，共培养 2 天后 hAMSCs 中 CK19、E-cad 和 Vimentin 的表达无明显差异。共培养 6 天后，hAMSCs 中 CK19、E-cad 和 Vimentin 的表达明显改变。组织学检测显示 hAMSCs 和 hAMSCs-CM 组子宫内膜腺体数量增加，纤维化面积显著减少。但两组间差异无统计学意义。体内荧光成像显示 GFP-hAMSCs 定位于子宫内膜基质，并逐渐发生凋亡。H&E 染色、MASSON 染色和 IHC 评估显示，hAMSCs 经阴道注射的效果与宫腔内注射相当。
我们的数据表明，hAMSCs 通过旁分泌促进子宫内膜修复，而不是通过转分化。
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