Center of Reproductive Medicine, The First Affiliated Hospital of Guangxi Medical University, Nanning, China.
Reproductive and Genetic Hospital, The First Affiliated Hospital of University of Science and Technology of China (USTC), Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China.
Biol Reprod. 2023 Dec 11;109(6):918-937. doi: 10.1093/biolre/ioad102.
Intrauterine adhesions (IUA) are a common gynecological problem. Stem cell therapy has been widely used in the treatment of IUA. However, due to the complex and harsh microenvironment of the uterine cavity, the effectiveness of such therapy is greatly inhibited. This study aimed to investigate whether melatonin pretreatment enhances the efficacy of human umbilical cord mesenchymal stem cells (HucMSCs) in IUA treatment in rats. First, we explored the effect of melatonin on the biological activity of HucMSCs in vitro through a macrophage co-culture system, Cell Counting Kit 8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (EdU), flow cytometry, immunofluorescence staining, and qRT-PCR. Subsequently, we established the IUA rat model and tracked the distribution of HucMSCs in this model. In addition, we observed the number of M1 and M2 macrophages through immunofluorescence staining and detected the levels of inflammatory cytokines. Four weeks after cell transplantation, HE, Masson, and immunohistochemical staining were performed. In vitro experiments showed that melatonin pretreatment of HucMSCs promoted proliferation, reduced apoptosis, up-regulated the stemness gene, and regulated macrophage polarization. In vivo, melatonin pretreatment caused more HucMSCs to remain in the uterine cavity. Melatonin-pretreated HucMSCs recruited more macrophages, regulated macrophage polarization, and reduced inflammation. Melatonin-pretreated HucMSCs relieved fibrosis, increased endometrium thickness, and up-regulated CD34, vimentin, proliferating cell nuclear antigen (PCNA), and alpha small muscle antigen (α-SMA) expression. Fertility tests showed that melatonin-pretreated HucMSCs increased the number of embryos. In summary, pretreatment with melatonin was beneficial for HucMSC treatment because it enhanced the cell's ability to recruit macrophages and regulate macrophage polarization, which led to the regeneration of the endometrium and improved pregnancy outcomes.
宫腔粘连（IUA）是一种常见的妇科问题。干细胞治疗已广泛应用于 IUA 的治疗。然而，由于宫腔内复杂和恶劣的微环境，这种治疗的效果受到了极大的抑制。本研究旨在探讨褪黑素预处理是否增强人脐带间充质干细胞（HucMSCs）在大鼠 IUA 治疗中的疗效。首先，我们通过巨噬细胞共培养系统、细胞计数试剂盒（CCK-8）、5-乙炔基-2'-脱氧尿苷（EdU）、流式细胞术、免疫荧光染色和 qRT-PCR 等方法探讨了褪黑素对 HucMSCs 体外生物学活性的影响。随后，我们建立了 IUA 大鼠模型，并跟踪了该模型中 HucMSCs 的分布。此外，我们通过免疫荧光染色观察了 M1 和 M2 巨噬细胞的数量，并检测了炎症细胞因子的水平。细胞移植 4 周后，进行 HE、Masson 和免疫组织化学染色。体外实验表明，褪黑素预处理 HucMSCs 可促进增殖，减少凋亡，上调干细胞基因，并调节巨噬细胞极化。在体内，褪黑素预处理使更多的 HucMSCs 留在子宫腔中。褪黑素预处理的 HucMSCs 募集了更多的巨噬细胞，调节了巨噬细胞极化，并减少了炎症。褪黑素预处理的 HucMSCs 缓解了纤维化，增加了子宫内膜厚度，并上调了 CD34、波形蛋白、增殖细胞核抗原（PCNA）和α-平滑肌肌动蛋白（α-SMA）的表达。生育能力测试表明，褪黑素预处理的 HucMSCs 增加了胚胎数量。综上所述，褪黑素预处理有利于 HucMSC 治疗，因为它增强了细胞募集巨噬细胞和调节巨噬细胞极化的能力，从而促进了子宫内膜的再生，改善了妊娠结局。
