Department of Obstetrics and Gynecology, Cathay General Hospital, Taipei, Taiwan.
Department of Obstetrics and Gynecology, School of Medicine, College of Medicine, Taipei Medical University, Taipei City, Taiwan.
PLoS One. 2019 Jan 29;14(1):e0210765. doi: 10.1371/journal.pone.0210765. eCollection 2019.
Adenomyosis is a medical condition defined by the abnormal presence of endometrial tissue within the myometrium, in which fibrosis occurs with new collagen deposition and myofibroblast differentiation. In this study, the effect of several mediators and growth factors on collagen expression was investigated on human endometrial stromal cells (fibroblasts) derived from adenomyotic endometrium.
RT-PCR, Western blot analysis, pharmacological interventions and siRNA interference were applied to primary cultured human endometrial stromal cells (fibroblasts). Immunohistochemistry was used to analyze protein expression in adenomyotic endometrium tissue specimens.
Of the tested mediators, transforming growth factor β1 (TGFβ1) and its isoforms were effective to induce collagen and connective tissue growth factor (CTGF) expression. Collagen and CTGF induction by TGFβ1 could be reduced by the inhibitors targeting DNA transcription, protein translation, and Smad2/3 signaling. Interestingly, TGFβ1 induced Smad2/3 phosphorylation and CTGF mRNA expression, but not collagen mRNA expression, suggesting that TGFβ1 mediates collagen expression through CTGF induction and Smad2/3 activation. In parallel, TGFβ1 and CTGF also induced expression of heat shock protein (HSP) 47, a protein required for the synthesis of several types of collagens. However, only CTGF siRNA knockdown, could compromise TGFβ1-induced collagen expression. Finally, the immunohistochemistry revealed vimentin- and α-SMA-positive staining for (myo)fibroblasts, TGFβ1, collagen, and CTGF in the subepithelial stroma region of human adenomyotic endometria.
We reveal here that TGFβ1, collagen, and CTGF are expressed in the stroma of adenomyotic endometria and demonstrate that TGFβ1 can induce collagen production in endometrium-derived fibroblasts through cellular Smad2/3-dependent signaling pathway and CTGF expression, suggesting that endometrial TGFβ may take part in the pathogenesis of adenomyosis and ectopic endometrium may participate in uterine adenomyosis.
子宫腺肌病是一种定义为子宫内膜组织异常存在于子宫肌层的医学病症，其中纤维化发生伴随着新的胶原沉积和肌成纤维细胞分化。在这项研究中，研究了几种介质和生长因子对来源于腺肌病子宫内膜的人子宫内膜基质细胞（成纤维细胞）胶原表达的影响。
应用 RT-PCR、Western blot 分析、药理学干预和 siRNA 干扰对原代培养的人子宫内膜基质细胞（成纤维细胞）进行处理。免疫组织化学用于分析腺肌病子宫内膜组织标本中的蛋白表达。
在所测试的介质中，转化生长因子β1（TGFβ1）及其同工型有效诱导胶原和结缔组织生长因子（CTGF）表达。针对 DNA 转录、蛋白质翻译和 Smad2/3 信号的抑制剂可降低 TGFβ1 诱导的胶原和 CTGF 表达。有趣的是，TGFβ1 诱导 Smad2/3 磷酸化和 CTGF mRNA 表达，但不诱导胶原 mRNA 表达，表明 TGFβ1 通过 CTGF 诱导和 Smad2/3 激活介导胶原表达。平行地，TGFβ1 和 CTGF 还诱导热休克蛋白（HSP）47 的表达，HSP47 是几种类型胶原合成所必需的蛋白。然而，只有 CTGF siRNA 敲低可损害 TGFβ1 诱导的胶原表达。最后，免疫组织化学显示上皮下基质区域的腺肌病子宫内膜中存在（肌）成纤维细胞、TGFβ1、胶原和 CTGF 的波形蛋白和 α-SMA 阳性染色。
我们在此揭示 TGFβ1、胶原和 CTGF 在腺肌病子宫内膜的基质中表达，并表明 TGFβ1 可通过细胞 Smad2/3 依赖性信号通路和 CTGF 表达诱导子宫内膜源性成纤维细胞产生胶原，提示子宫内膜 TGFβ 可能参与子宫腺肌病的发病机制，异位子宫内膜可能参与子宫腺肌病的发生。
