Bone marrow mesenchymal stem cells combined with estrogen synergistically promote endometrial regeneration and reverse EMT via Wnt/β-catenin signaling pathway.
Bone marrow mesenchymal stem cells combined with estrogen synergistically promote endometrial regeneration and reverse EMT via Wnt/β-catenin signaling pathway.
复制标题
骨髓间充质干细胞联合雌激素协同促进子宫内膜再生并通过Wnt/β-catenin信号通路逆转EMT
DOI：
10.1186/s12958-022-00988-1
复制
发表时间：
2022-08-15
影响因子：
4.4
通讯作者：
Liu, Dan
中科院分区：
文献类型：
--
作者：
Yuan, Liwei;Cao, Jia;Hu, Mingyue;Xu, Dabao;Li, Yan;Zhao, Shiyun;Yuan, Juanjuan;Zhang, Huixing;Huang, Yani;Jin, He;Chen, Meixia;Liu, Dan
关键词：
宫腔粘连（IUA）是一种以子宫内膜基底层损伤导致宫腔闭塞为特征的临床疾病。骨髓间充质干细胞（BMSCs）移植主要通过旁分泌能力具有促进子宫内膜再生的潜力。雌激素是子宫内膜损伤修复中不可或缺的重要因素，据报道，雌激素在干细胞移植治疗中是一种有前景的辅助治疗应用。本研究旨在探讨BMSCs与雌激素在兔IUA双重损伤模型中促进子宫内膜再生、恢复子宫内膜形态的协同作用及其分子机制。通过成脂和成骨分化以及流式细胞术测定分离和鉴定 BMSC。采用机械刮除和脂多糖感染双重损伤方法建立兔IUA动物模型。此外，我们还研究了骨髓间充质干细胞和雌激素单独或组合在兔模型中的治疗作用。采用体内荧光成像观察PKH26标记的BMSCs的保留情况。分别采用H&E和Masson染色观察子宫内膜腺体的数量和纤维化程度。采用Western blotting、免疫组织化学和免疫荧光染色检测与子宫内膜上皮、子宫内膜纤维化和EMT相关的生物标志物。最后通过Western blotting检测Wnt/β-catenin通路核心分子的蛋白表达量。 PKH26标记的荧光结果显示，当原位移植到子宫腔中时，BMSCs出现并定位于子宫内膜腺体和细胞外基质区域。组织学检测显示BMSCs联合雌激素治疗组子宫内膜腺体数量明显增加，子宫内膜纤维化面积明显减少。此外，在联合组中发现纤维化标志物（纤连蛋白、胶原蛋白 I、a-SMA）和间质标志物（ZEB1、波形蛋白、N-钙粘蛋白）表达下调，以及 E-钙粘蛋白表达上调。进一步的体内染色研究发现，联合组中CK7的荧光强度比直接BMSC宫内移植的荧光强度更强，而vimentin则表现出相反的结果。此外，BMSCs联合雌激素组Wnt/β-catenin信号通路的β-catenin、Axin2、C-myc、CycinE蛋白水平较其他治疗组升高。 BMSCs联合雌激素可促进干细胞分化为子宫内膜上皮细胞，有利于受损子宫内膜的再生。协同作用的潜在机制可能是通过激活Wnt/β-catenin信号通路抑制EMT的发生。在线版本包含可在 10.1186/s12958-022-00988-1 获取的补充材料。
Intrauterine adhesion (IUA) is a clinical disease characterized by the uterine cavity occlusion caused by the damage of the endometrial basal layer. Bone marrow mesenchymal stem cells (BMSCs) transplantation have the potential to promote endometrial regeneration mainly through paracrine ability. Estrogen is an indispensable and important factor in the repair of endometrial damage, which has been reported as a promising and adjunctive therapeutic application for stem cell transplantation therapy. This study aims to investigate the synergistic effect of BMSCs and estrogen on improving the endometrial regeneration and restoring the endometrium morphology in a dual damage model of IUA in rabbits and the underlying molecular mechanisms. BMSCs were isolated and identified by adipogenic and osteogenic differentiation and flow cytometry assays. The rabbit IUA animal model was established by a dual damage method of mechanical curettage and lipopolysaccharide infection. Additionally, we investigated the therapeutic impact of both BMSCs and estrogen either separately or in combination in a rabbit model. The retention of PKH26-labeled BMSCs was observed by vivo fluorescence imaging.The number of endometrial glands and the degree of fibrosis were observed by H&E and Masson staining respectively. Western blotting, Immunohistochemistry and immunofluorescence staining were performed to detect biomarkers related to endometrial epithelium, endometrial fibrosis and EMT. Finally, the protein expression of core molecules of Wnt/β-catenin pathway was detected by Western blotting. PKH26-labeled fluorescence results revealed that BMSCs appeared and located in the endometrial glands and extracellular matrix area when orthotopic transplanted into the uterine cavity. Histological assays showed that remarkably increasing the number of endometrial glands and decreasing the area of endometrial fibrosis in the BMSCs combined with estrogen treatment group. Moreover, downregulated expression of fibrosis markers (fibronectin, CollagenI, a-SMA) and interstitial markers (ZEB1, Vimentin, N-cadherin), as well as upregulated E-cadherin expression were found in the combined group. Further study of in vivo staining revealed that fluorescence intensity of CK7 was stronger in the combined group than that of direct BMSCs intrauterine transplantation, while vimentin showed the opposite results. Moreover, the protein levels of β-catenin, Axin2, C-myc, CycinE of Wnt/β-catenin signaling pathway increased in the BMSCs combined with estrogen group than in the other treatment groups. BMSCs combined with estrogen can promote the differentiation of stem cells into endometrial epithelial cells to facilitate the regeneration of damaged endometrium. The potential mechanism of the synergistic effect may inhibit the occurrence of EMT by activating the Wnt/β-catenin signaling pathway. The online version contains supplementary material available at 10.1186/s12958-022-00988-1.
登录
查看更多内容
影响因子：
4
作者：
Du, Hongling;Naqvi, Hanyia;Taylor, Hugh S.
通讯作者：
Taylor, Hugh S.
影响因子：
6.8
作者：
Li, Changjiang;Cai, Aiqi;Wang, Jia
通讯作者：
Wang, Jia
影响因子：
8
作者：
Buschhaus, Johanna M.;Rajendran, Shrila;Humphries, Brock A.;Cutter, Alyssa C.;Muniz, Ayse J.;Ciavattone, Nicholas G.;Buschhaus, Alexander M.;Caneque, Tatiana;Nwosu, Zeribe C.;Sahoo, Debashis;Bevoor, Avinash S.;Shah, Yatrik M.;Lyssiotis, Costas A.;Ghosh, Pradipta;Wicha, Max S.;Rodriguez, Raphael;Luker, Gary D.
通讯作者：
Luker, Gary D.
Notch 信号传导的抑制通过自噬激活和 PTEN-PI3K/AKT/mTOR 途径促进间充质干细胞的脂肪形成分化
影响因子：
--
作者：
Song, Bao-quan;Chi, Ying;Han, Zhong-chao
通讯作者：
Han, Zhong-chao
影响因子：
--
作者：
Wang, Jianmei;Ju, Baohui;Zhang, Yujuan
通讯作者：
Zhang, Yujuan