Endometriotic mesenchymal stem cells significantly promote fibrogenesis in ovarian endometrioma through the Wnt/β-catenin pathway by paracrine production of TGF-β1 and Wnt1
Endometriotic mesenchymal stem cells significantly promote fibrogenesis in ovarian endometrioma through the Wnt/β-catenin pathway by paracrine production of TGF-β1 and Wnt1
复制标题
子宫内膜异位间充质干细胞通过旁分泌产生TGF-β1和Wnt1，通过Wnt/β-连环蛋白途径显着促进卵巢子宫内膜异位瘤的纤维化
DOI：
10.1093/humrep/dew058
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发表时间：
2016-06-01
影响因子：
6.1
通讯作者：
Zhang, Songying
中科院分区：
文献类型：
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作者：
Li, Jing;Dai, Yongdong;Zhang, Songying
子宫内膜异位间充质干细胞(ECTO-MSCs)是否参与了卵巢子宫内膜异位症的纤维化过程？ECTO-MSCs通过分泌转化生长因子-β1(TGF-β1)和WNT1，通过Wnt/β-catenin途径增强卵巢子宫内膜异位内膜间质细胞的纤维化行为。子宫内膜异位症，尤其是卵巢子宫内膜异位症相关纤维化的发病机制尚不清楚。我们分析了15例月经周期正常的育龄卵巢子宫内膜异位症患者的子宫内膜标本。54只裸鼠一次性注射来自14名非子宫内膜异位症患者的增殖期子宫内膜碎片，条件培养液来自子宫内膜间充质干细胞(Euto-MSCs)和EECO-MSCs。用细胞计数试剂盒8、Transwell和胶原胶收缩实验观察CM对卵巢子宫内膜异位内膜和异位内膜间质细胞增殖、迁移、侵袭和胶原凝胶收缩的影响。采用实时定量逆转录-聚合酶链式反应和免疫印迹法检测CM对Euto-ESCs和Ecoto-ESCs中纤维化标志物[包括α-平滑肌肌动蛋白、I型胶原、结缔组织生长因子和纤维连接蛋白(FN)]表达的影响。此外，利用免疫缺陷裸鼠子宫内膜异位症移植模型，分析了外源性间充质干细胞对异位内膜植入物的促纤维化作用。结果表明，与对照组和内异症间充质干细胞相比，外源性间充质干细胞明显促进卵巢子宫内膜异位内异症患者外胚层和外胚间充质干细胞的增殖、迁移、侵袭和胶原凝胶收缩。经ECTO-MSC CM治疗后，Euto-ESCs和Ecto-ESCs中纤维化标志物的表达水平显著升高。ECTO-MSCs较Euto-MSCs分泌更高水平的转化生长因子-β1和WNT1。此外，转化生长因子β1和WNT1均显著增加Euto-ESCs和ECTO-ESCs中纤维化标志物的表达，这一作用可被抗转化生长因子-β1抗体或Wnt1负调控因子Dickkopf相关蛋白1(Dkk1)逆转。机制研究表明，ECTO-MSC CM激活了基质细胞中的Wnt/β-catenin信号通路。动物实验表明，转化生长因子-β1和WNT1以及ECTO-MSC CM显著增加FN和I型胶原的表达，从而促进子宫内膜异位症纤维化的进展。然而，许多其他生长因子和细胞类型也可能参与了发病机制。因此，需要进一步研究卵巢子宫内膜异位内膜细胞的旁分泌作用。卵巢内膜间充质干细胞可能参与卵巢内膜纤维化的发病机制。本研究得到国家自然科学基金中国(81471505-81270657)的部分资助。没有宣布任何相互竞争的利益。
Are endometriotic mesenchymal stem cells (Ecto-MSCs) involved in the fibrosis of ovarian endometrioma?Ecto-MSCs enhanced the fibrotic behavior of stromal cells in ovarian endometrioma through the Wnt/beta-catenin pathway by paracrine production of transforming growth factor-beta 1 (TGF-beta 1) and Wnt1.Endometriosis is characterized by ectopic outgrowth of endometrial stroma and glands surrounded by dense fibrous tissues. The pathogenesis of endometriosis, especially ovarian endometrioma-associated fibrosis, is still unknown.We analyzed endometrial samples from 15 patients of reproductive age with ovarian endometrioma and normal menstrual cycles. A total of 54 nude mice received a single injection of proliferative endometrial fragments from 14 individuals without endometriosis.Conditioned medium (CM) was collected from endometrial mesenchymal stem cells (Euto-MSCs) and Ecto-MSCs. The effects of CM on cell proliferation, migration, invasion and collagen gel contraction of endometrial and endometriotic stromal cells (Euto- and Ecto-ESCs) in ovarian endometrioma were evaluated by cell counting kit-8, transwell and collagen gel contraction assays. Effects of CM on fibrotic markers' expression [including alpha-smooth muscle actin, Type I collagen, connective tissue growth factor and fibronectin (FN)] in Euto- and Ecto-ESCs were determined by real-time reverse-transcription-polymerase chain reaction and western blotting. Additionally, fibrogenic effects of Ecto-MSC CM treatment on endometriotic implants were analyzed using a xenograft model of endometriosis in immunodeficient nude mice.Our results demonstrated that Ecto-MSC CM significantly promoted cell proliferation, migration, invasion and collagen gel contraction of Euto- and Ecto-ESCs from patients with ovarian endometrioma compared with control and Euto-MSC CM. Expression levels of fibrotic markers in Euto- and Ecto-ESCs were dramatically elevated after treatment with Ecto-MSC CM. Ecto-MSCs secreted higher levels of TGF-beta 1 and Wnt1 compared with Euto-MSCs. Furthermore, both TGF-beta 1 and Wnt1 significantly increased expression of fibrotic markers in Euto- and Ecto-ESCs, which was reversed by an anti-TGF-beta 1 antibody or Wnt1 negative regulator, Dickkopf-related protein 1 (Dkk1). Mechanistic studies demonstrated that Wnt/beta-catenin signaling pathways in stromal cells were activated by Ecto-MSC CM. Animal experiments showed that TGF-beta 1 and Wnt1 as well as Ecto-MSC CM markedly increased the expression of FN and collagen I, which enhanced the progression of fibrosis in endometriosis.To our knowledge, this is the first study to identify the role of Ecto-MSCs in the pathogenesis of fibrosis in ovarian endometrioma. However, numerous other growth factors and cell types may also be involved in the pathogenesis. Therefore, further studies are required to elucidate the paracrine effects of cells in ovarian endometrioma.Ecto-MSCs may be involved in the pathogenesis of fibrosis in ovarian endometrioma.This study was supported in part by the National Natural Science Foundation of China (81471505 and 81270657). No competing interests are declared.