Department of Orthopedics and Trauma Surgery, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University, 40225 Düsseldorf, Germany.
Int J Mol Sci. 2023 Oct 11;24(20):15097. doi: 10.3390/ijms242015097.
Dupuytren's disease (DD) is a fibroproliferative disorder affecting the palmar fascia, causing functional restrictions of the hand and thereby limiting patients' daily lives. The disturbed and excessive myofibroblastogenesis, causing DD, is mainly induced by transforming growth factor (TGF)-β1. But, the extent to which impaired TGF-β1 release or TGF-β signal degradation is involved in pathologically altered myofibroblastogenesis in DD has been barely examined. Therefore, the complex in which TGF-β1 is secreted in the extracellular matrix to elicit its biological activity, and proteins such as plasmin, integrins, and matrix metalloproteinases (MMPs), which are involved in the TGF-β1 activation, were herein analyzed in DD-fibroblasts (DD-FBs). Additionally, TGF-β signal degradation via caveolin-1 was examined with 5-fluoruracil (5-FU) in detail. Gene expression analysis was performed via Western blot, PCR, and immunofluorescence analyses. As a surrogate parameter for disturbed myofibroblastogenesis, 𝛼-smooth-muscle-actin (𝛼-SMA) expression was evaluated. It was demonstrated that latency-associated peptide (LAP)-TGF-β and latent TGF-β-binding protein (LTBP)-1 involved in TGF-β-complex building were significantly upregulated in DD. Plasmin a serinprotease responsible for the TGF-β release was significantly downregulated. The application of exogenous plasmin was able to inhibit disturbed myofibroblastogenesis, as measured via 𝛼-SMA expression. Furthermore, a reduced TGF-β1 degradation was also involved in the pathological phenotype of DD, because caveolin-1 expression was significantly downregulated, and if rescued, myofibroblastogenesis was also inhibited. Therefore, our study demonstrates that a deficient release and degradation of TGF-β1 are important players in the pathological phenotype of DD and should be addressed in future research studies to improve DD therapy or other related fibrotic conditions.
掌腱膜挛缩症(Dupuytren's disease,DD)是一种影响掌筋膜的纤维增生性疾病,导致手部功能受限,从而限制了患者的日常生活。导致 DD 的失调和过度肌成纤维细胞生成主要由转化生长因子(transforming growth factor,TGF)-β1 诱导。但是,在 DD 病理性改变的肌成纤维细胞生成中,受损的 TGF-β1 释放或 TGF-β信号降解的程度几乎没有被研究过。因此,本文分析了 TGF-β1 在细胞外基质中分泌并发挥其生物学活性的复合物,以及纤溶酶、整合素和基质金属蛋白酶(matrix metalloproteinases,MMPs)等参与 TGF-β1 激活的蛋白质,在 DD 成纤维细胞(fibroblasts,DD-FBs)中的作用。此外,还详细研究了通过小窝蛋白-1(caveolin-1)对 TGF-β信号的降解。通过 Western blot、PCR 和免疫荧光分析进行基因表达分析。作为肌成纤维细胞生成失调的替代参数,评估了 α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的表达。结果表明,Latency-associated peptide(LAP)-TGF-β和参与 TGF-β 复合物构建的Latent TGF-β-binding protein(LTBP)-1在 DD 中显著上调。丝氨酸蛋白酶纤溶酶,负责 TGF-β 的释放,显著下调。外源性纤溶酶的应用能够抑制失调的肌成纤维细胞生成,如通过 α-SMA 表达来衡量。此外,TGF-β1 降解的减少也与 DD 的病理表型有关,因为小窝蛋白-1 的表达显著下调,如果得到挽救,肌成纤维细胞生成也会受到抑制。因此,我们的研究表明,TGF-β1 的释放和降解不足是 DD 病理表型的重要因素,应该在未来的研究中加以解决,以改善 DD 治疗或其他相关纤维化疾病。