Key Laboratory of Reproductive Dysfunction Diseases and Fertility Remodeling of Liaoning Province, Reproductive Medicine Center, Obstetrics and Gynecology Department, Shengjing Hospital affiliated to China Medical University, No. 39 Huaxiang Road, Tiexi District, Shenyang, Liaoning, China.
Key Laboratory of Health Ministry for Congenital Malformation, Shengjing Hospital affiliated to China Medical University, No. 7, Economic Development Zone, Benxi, Liaoning, China.
Stem Cell Res Ther. 2019 Feb 15;10(1):61. doi: 10.1186/s13287-019-1155-7.
Intrauterine adhesion (IUA) is a major cause of female secondary infertility. We previously demonstrated that menstrual blood-derived stromal cell (MenSC) transplantation helped severe IUA patients have pregnancy and endometrium regeneration. We also initiated platelet-rich plasma (PRP) acted as a beneficial supplement in MenSC culturing and a potential endometrial receptivity regulator. Here, we investigated the therapeutic effect of combined transplantation of MenSCs with PRP in rat IUA models and the mechanisms of MenSCs in endometrium regeneration.
Rat IUA models were established by intrauterine mechanical injured. Nine days later, all rats were randomly assigned to four groups received different treatment: placebo, MenSC transplantation, PRP transplantation, and MenSCs + PRP transplantation. The traces of MenSCs were tracked with GFP label. Endometrial morphology and pathology, tissue proliferation, inflammation, pregnancy outcomes, and mechanism of MenSCs in the regeneration of endometrium were investigated.
Notably, at days 9 and 18 post-treatment, MenSC transplantation significantly improved endometrial proliferation, angiogenesis, and morphology recovery and decreased collagen fibrosis and inflammation in the uterus. MenSCs had lesion chemotaxis, colonized around the endometrial glands. Gene expression of human-derived secretory protein IGF-1, SDF-1, and TSP-1 was detected in the uterus received MenSCs at day 18. The three treatments can all improve fertility in IUA rats. Moreover, gene expressions of cell proliferation, developmental processes, and other biological processes were induced in MenSC transplantation group. Hippo signaling pathway was the most significantly changed pathway, and the downstream factors CTGF, Wnt5a, and Gdf5 were significantly regulated in treatment groups. PRP enhanced these parameters through a synergistic effect.
In summary, MenSCs could effectively improve uterine proliferation, markedly accelerate endometrial damage repairment and promote fertility restoration in IUA rats, suggesting a paracrine restorative effect and Hippo signaling pathway stimulation. Our results indicate MenSCs, a valuable source of cells for transplantation in the treatment intrauterine adhesion. Combined with PRP, this cell therapy was more effective.
宫腔粘连（IUA）是女性继发性不孕的主要原因。我们之前的研究表明，经血来源的基质细胞（MenSC）移植有助于严重 IUA 患者妊娠和子宫内膜再生。我们还发现富含血小板的血浆（PRP）在 MenSC 培养中作为一种有益的补充，并可能是子宫内膜容受性调节剂。在这里，我们研究了 MenSCs 与 PRP 联合移植在大鼠 IUA 模型中的治疗效果以及 MenSCs 在子宫内膜再生中的机制。
通过宫腔内机械损伤建立大鼠 IUA 模型。9 天后，所有大鼠被随机分为四组，分别接受不同的治疗：安慰剂、MenSC 移植、PRP 移植和 MenSCs+PRP 移植。通过 GFP 标记跟踪 MenSCs 的踪迹。研究了子宫内膜形态和病理学、组织增殖、炎症、妊娠结局以及 MenSCs 在子宫内膜再生中的机制。
值得注意的是，在治疗后 9 天和 18 天，MenSC 移植显著改善了子宫内膜增殖、血管生成和形态恢复，并减少了子宫内的胶原纤维化和炎症。MenSCs 具有病变趋化性，定植在子宫内膜腺周围。在接受 MenSCs 的子宫中，在第 18 天检测到人源性分泌蛋白 IGF-1、SDF-1 和 TSP-1 的基因表达。三种治疗方法都可以改善 IUA 大鼠的生育能力。此外，在 MenSC 移植组中诱导了细胞增殖、发育过程等生物学过程的基因表达。Hippo 信号通路是变化最显著的通路，下游因子 CTGF、Wnt5a 和 Gdf5 在治疗组中受到显著调节。PRP 通过协同作用增强了这些参数。
总之，MenSCs 可有效改善子宫增殖，显著加速子宫内膜损伤修复，促进 IUA 大鼠生育力恢复，提示存在旁分泌修复作用和 Hippo 信号通路刺激。我们的研究结果表明，MenSCs 是一种有价值的细胞移植治疗宫腔粘连的来源。与 PRP 联合使用，这种细胞疗法更有效。
