人脐带血间充质干细胞向子宫内膜细胞转分化并通过 NF-κB 信号通路调节兔宫腔粘连内膜中的 Th17/Treg 平衡。

Human umbilical cord blood-derived MSCs trans-differentiate into endometrial cells and regulate Th17/Treg balance through NF-κB signaling in rabbit intrauterine adhesions endometrium.

机构信息

Department of Obstetrics and Gynecology, Zhengzhou Central Hospital Affiliated to Zhengzhou University, Zhengzhou, Henan, People's Republic of China.

Branch Center of Advanced Medical Research Center, Zhengzhou Central Hospital Affiliated to Zhengzhou University, Zhengzhou, Henan, People's Republic of China.

出版信息

Stem Cell Res Ther. 2022 Jul 15;13(1):301. doi: 10.1186/s13287-022-02990-1.

DOI:10.1186/s13287-022-02990-1

PMID:35841027

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9284747/

Abstract

PURPOSE

The fundamental cause of intrauterine adhesions (IUAs) is the destruction and reduction in stem cells in endometrial basal layer, resulting in endometrial reconstruction very difficult. The purpose of this study was to investigate the effects and underlying mechanism of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) on the endometrial reconstruction after transplantation.

METHODS

hUCB-MSCs were isolated and identified by flow cytometry, osteogenic, adipogenic and chondrogenic differentiation assays. The rabbit IUA models were established and set five groups (control, 14/28th day after surgery, estrogen and hUCB-MSCs treatment). The number of endometrial glands and the fibrosis rate were evaluated using HE and Masson staining, respectively. Endometrial proliferation, angiogenesis and inflammation were evaluated by immunohistochemical staining of ER, Ki-67and TGF-β1, respectively. Single-cell RNA sequencing (scRNA-seq) was applied to explore the cell differentiation trajectory after hUCB-MSCs transplanted into IUA endometrium. Finally, molecular mechanism of hUCB-MSCs repairing damaged endometrium was investigated by RNA sequencing, qRT-PCR and Western blot assays.

RESULTS

After transplantation of the hUCB-MSCs, the increase in endometrial gland number, estrogen receptor (ER) and Ki-67 expression, and the decrease in fibrosis rate and TGF-β expression (P < 0.05), suggested the endometrial repair, angiogenesis and inflammatory suppression. The therapeutic effect of hUCB-MSCs was significantly improved compared with 28th day after surgery and estrogen group. ScRNA-seq demonstrated that the transplanted hUCB-MSCs can trans-differentiate into endometrial cells: epithelial, fibroblast and macrophage. RNA sequencing of six IUA samples combined with qRT-PCR and Western blot assays further revealed that hUCB-MSCs may regulate Th17/Treg balance through NF-κB signaling, thus inhibiting the immune response of damaged endometrium.

CONCLUSIONS

Our study demonstrated that hUCB-MSCs can repair damaged endometrium through trans-differentiation, immunomodulatory capacities and NF-κB signaling, suggesting the treatment value of hUCB-MSCs in IUA.

摘要

目的

宫腔粘连（IUAs）的根本原因是子宫内膜基底层干细胞的破坏和减少，导致子宫内膜重建非常困难。本研究旨在探讨人脐带来源间充质干细胞（hUCB-MSCs）移植后对子宫内膜重建的影响及其潜在机制。

方法

采用流式细胞术分离鉴定 hUCB-MSCs，进行成骨、成脂和成软骨分化鉴定。建立兔 IUA 模型，设 5 组（对照组、术后 14 天、28 天、雌激素和 hUCB-MSCs 治疗组）。采用 HE 和 Masson 染色分别评估子宫内膜腺数量和纤维化率。采用 ER、Ki-67 和 TGF-β1 的免疫组化染色分别评估子宫内膜增殖、血管生成和炎症。应用单细胞 RNA 测序（scRNA-seq）探索 hUCB-MSCs 移植入 IUA 子宫内膜后的细胞分化轨迹。最后，通过 RNA 测序、qRT-PCR 和 Western blot 分析研究 hUCB-MSCs 修复受损子宫内膜的分子机制。

结果

hUCB-MSCs 移植后，子宫内膜腺数量增加，ER 和 Ki-67 表达增强，纤维化率和 TGF-β表达降低（P<0.05），提示子宫内膜修复、血管生成和炎症抑制。与术后 28 天和雌激素组相比，hUCB-MSCs 的治疗效果明显改善。scRNA-seq 表明，移植的 hUCB-MSCs 可向子宫内膜细胞（上皮细胞、成纤维细胞和巨噬细胞）转分化。对 6 个 IUA 样本的 RNA 测序结合 qRT-PCR 和 Western blot 分析进一步表明，hUCB-MSCs 可能通过 NF-κB 信号调节 Th17/Treg 平衡，从而抑制受损子宫内膜的免疫反应。