Transforming growth factor β1 signaling coincides with epithelial-mesenchymal transition and fibroblast-to-myofibroblast transdifferentiation in the development of adenomyosis in mice
Transforming growth factor β1 signaling coincides with epithelial-mesenchymal transition and fibroblast-to-myofibroblast transdifferentiation in the development of adenomyosis in mice
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DOI：
10.1093/humrep/dev314
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发表时间：
2016-02-01
影响因子：
6.1
通讯作者：
Guo, Sun-Wei
中科院分区：
文献类型：
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作者：
Shen, Minhong;Liu, Xishi;Guo, Sun-Wei
血小板在子宫腺肌病的发展中有作用吗？活化的血小板与转化生长因子（TGF）-β 1的释放和TGF-β/Smad信号传导途径的诱导以及子宫腺肌病小鼠模型中上皮-间充质转化（EMT）和成纤维细胞向肌成纤维细胞转分化（FMT）的证据一致，最终导致纤维化，EMT和FMT在子宫内膜异位症中起重要作用。EMT与子宫腺肌病的发生有关，但这主要是基于横断面观察。目前尚不清楚子宫腺肌病病变及其微环境是否具有促进EMT和FMT的机制，最终导致纤维化。目前还没有任何关于血小板在子宫腺肌病发展中的作用的研究，即使子宫腺肌病病变经历反复的组织损伤和修复周期，这涉及血小板的参与，并构成了一个环境有利于fibrogenes.Adenomyosis诱导28只雌性ICR小鼠的新生儿剂量的他莫昔芬。另外32名患者在没有他莫昔芬的情况下接受了腹腔给药。分别于5、10、15、42和60日龄分批处死有和无子宫腺肌病的ICR雌性小鼠。评估子宫内膜组织的肌层浸润深度，并在异位（对于子宫腺肌病小鼠）和在位（对照）子宫内膜组织样品中进行EMT和FMT的生物标志物以及TGF-β 1、磷酸化Smad 3（p-Smad 3）和增殖、血管生成和细胞外基质（ECM）沉积物的标志物的免疫组织化学分析。Masson三色染色和货车Gieson染色用于定量病变中纤维化的程度。虽然TGF-β 1在对照子宫内膜中的免疫反应性一直较低，但其水平从第10天开始急剧增加沿着血小板聚集程度也随之增加。随着子宫腺肌病的进展，TGF-β 1和p-Smad 3的染色逐渐增加，同时沿着增殖、血管生成和ECM沉积的标志物。一致地，波形蛋白（基质或间充质细胞的标记物）的染色也增加，而E-钙粘蛋白（上皮细胞的标记物）的染色减少。PR-B染色也逐渐减少。从第42天开始，α-SMA染色（肌成纤维细胞的标志物）在病变中升高，而在对照子宫内膜中为阴性。同时，纤维化的程度也增加了。这项研究受到组织化学和免疫组化分析的限制，缺乏干预。像他们的子宫内膜异位症对应，子宫腺肌病病变及其微环境可能包含所有必要的分子机制，以促进纤维化。血小板诱导的TGF-β/Smad信号通路的激活可能是子宫腺肌病发展中EMT和FMT的驱动力，导致纤维化。这项研究提供了第一个证据，证明子宫腺肌病病变是经历反复损伤和愈合的伤口，因此，血小板在子宫腺肌病的发展中起着关键作用。这表明了抗血小板治疗子宫腺肌病的潜力，也为开发新的诊断或预后目的的生物标志物开辟了新的途径。没有一个作者有什么要披露的。
Do platelets have any role in the development of adenomyosis?Activated platelets coincide with the release of transforming growth factor (TGF)-beta 1 and induction of the TGF-beta/Smad signaling pathway as well as evidence of epithelial-mesenchymal transition (EMT) and fibroblast-to-myofibroblast transdifferentiation (FMT) in a mouse model of adenomyosis, resulting ultimately in fibrosis, as in adenomyosis.Both EMT and FMT are known to play vital roles in fibrogenesis in general and in endometriosis in particular. EMT has been implicated in the development of adenomyosis, but this was based primarily on cross-sectional observation. It is unclear as to whether adenomyotic lesions and their microenvironment have the machinery to promote EMT and FMT, resulting ultimately in fibrosis. There has not been any published study on the role of platelets in the development of adenomyosis, even though adenomyotic lesions undergo repeated cycles of tissue injury and repair, which implicates the involvement of platelets and constitutes an environment conducive for fibrogenesis.Adenomyosis was induced in 28 female ICR mice by neonatal dosing of tamoxifen. Another 32 were neonatally dosed without tamoxifen. These mice were sacrificed serially and their tissue samples were subsequently evaluated.Female ICR mice with and without induced adenomyosis were sacrificed in batch at 5, 10, 15, 42 and 60 days of age. The depth of myometrial infiltration of endometrial tissues was assessed and immunohistochemistry analysis of biomarkers of EMT and FMT, as well as TGF-beta 1, phosphorylated Smad3 (p-Smad3) and markers of proliferation, angiogenesis and extracellular matrix (ECM) deposits was performed in ectopic (for adenomyotic mice) and eutopic (controls) endometrial tissue samples. Masson trichrome and Van Gieson stainings were performed to quantify the extent of fibrosis in lesions. Progesterone receptor isoform B (PR-B) staining also was performed.While TGF-beta 1 immunoreactivity was consistently low in control endometrium, its level was increased dramatically starting from Day 10, along with the extent of platelet aggregation. Staining for TGF-beta 1 and p-Smad3 increased progressively as adenomyosis progressed, along with markers for proliferation, angiogenesis and ECM deposits. Consistently, staining of vimentin (a marker for stromal or mesenchymal cells) was also increased while that of E-cadherin (a marker for epithelial cells) was reduced. PR-B staining also decreased progressively. Starting from Day 42, alpha-SMA staining, a marker for myofibroblasts, was elevated in lesions, while in control endometrium, it was negative. Concomitantly, the extent of fibrosis also was increased.This study is limited by the use of histochemistry and immunohistochemistry analyses only and the lack of intervention.Like their endometriotic counterpart, adenomyotic lesions and their microenvironment may contain all the necessary molecular machinery to promote fibrogenesis. Platelet-induced activation of the TGF-beta/Smad signaling pathway may be a driving force in EMT and FMT in the development of adenomyosis, leading to fibrosis. This study provides the first piece of evidence that adenomyotic lesions are wounds that undergo repeated injury and healing, and as such, platelets play critical roles in the development of adenomyosis. It suggests the potential for the use of anti-platelet therapy in the treatment of adenomyosis, and also opens a new venue for developing novel biomarkers for diagnostic or prognostic purposes.Support for data collection and analysis was provided by grants from the National Science Foundation of China. None of the authors has anything to disclose.