METHODS: Uterine injury was surgically induced in a rat model, followed by immediate transplantation of 5 × 10 ^ 5 hUC-MSCs to each side of the uterus. Uterine morphology was evaluated at days 14 and 30 using HE and Masson staining. Immunohistochemistry assessed macrophage polarization, angiogenesis and endometrial receptivity in the endometrium. Additionally, the regulatory effects of hUC-MSCs on macrophage polarization were explored through coculture. qRT-PCR quantified the expression of anti-inflammatory (IL10 and Arg1) and pro-inflammatory (iNOS and TNF-α) factors. Western blotting evaluated CD163 expression.
RESULTS: Transplantation of hUC-MSCs promoted the healing of uterine injuries and tissue regeneration while inhibiting tissue fibrosis. Immunohistochemistry at days 14 and 30 post-transplantation demonstrated the polarization of macrophages toward the M2 phenotype in the uterine injury area in the presence of hUC-MSCs. Furthermore, hUC-MSC transplantation improved angiogenesis and endometrial receptivity in the uterine injury rat model, associated with increased IL10 expression. hUC-MSC-induced angiogenesis can be resisted by depleted macrophages. In vitro coculture experiments further demonstrated that hUC-MSCs promoted IL10 expression in macrophages while suppressing TNF-α and iNOS expression. Western blotting showed enhanced CD163 expression in macrophages following hUC-MSC treatment.
CONCLUSIONS: hUC-MSCs contribute to the healing of uterine injuries by targeting macrophages to promote angiogenesis and the expression of anti-inflammatory factors.
方法:在大鼠模型中手术诱发子宫损伤,然后立即将5×10^5hUC-MSCs移植到子宫两侧。在第14天和第30天使用HE和Masson染色评估子宫形态。免疫组织化学评估巨噬细胞极化,子宫内膜血管生成和子宫内膜容受性。此外,通过共培养探讨hUC-MSCs对巨噬细胞极化的调节作用。qRT-PCR定量抗炎(IL10和Arg1)和促炎(iNOS和TNF-α)因子的表达。Western印迹评估CD163表达。
结果:hUC-MSCs移植促进子宫损伤愈合和组织再生,同时抑制组织纤维化。移植后第14天和第30天的免疫组织化学表明,在hUC-MSC的存在下,子宫损伤区域中巨噬细胞向M2表型的极化。此外,hUC-MSC移植改善子宫损伤大鼠模型血管生成和子宫内膜容受性,与IL10表达增加相关。hUC-MSC诱导的血管生成可被耗竭的巨噬细胞抵抗。体外共培养实验进一步证明,hUC-MSC促进巨噬细胞中IL10的表达,同时抑制TNF-α和iNOS的表达。Western印迹显示hUC-MSC处理后巨噬细胞中CD163表达增强。
结论:hUC-MSCs通过靶向巨噬细胞促进血管新生和抗炎因子的表达促进子宫损伤的愈合。