Howard Jeffrey C, Varallo Vincenzo M, Ross Douglas C, Roth James H, Faber Kenneth J, Alman Benjamin, Gan Bing Siang
Department of Surgery, University of Western Ontario, London, Ontario, Canada.
BMC Musculoskelet Disord. 2003 Jul 16;4:16. doi: 10.1186/1471-2474-4-16.
Dupuytren's contracture or disease (DD) is a fibro-proliferative disease of the hand that results in the development of scar-like, collagen-rich disease cords within specific palmar fascia bands. Although the molecular pathology of DD is unknown, recent evidence suggests that beta-catenin may play a role. In this study, collagen matrix cultures of primary disease fibroblasts show enhanced contraction and isometric tension-dependent changes in beta-catenin and fibronectin levels.
Western blots of beta-catenin and fibronectin levels were determined for control and disease primary cell cultures grown within stressed- and attached-collagen matrices. Collagen contraction was quantified, and immunocytochemistry analysis of filamentous actin performed.
Disease cells exhibited enhanced collagen contraction activity compared to control cells. Alterations in isometric tension of collagen matrices triggered dramatic changes in beta-catenin and fibronectin levels, including a transient increase in beta-catenin levels within disease cells, while fibronectin levels steadily decreased to levels below those seen in normal cell cultures. In contrast, both fibronectin and beta-catenin levels increased in attached collagen-matrix cultures of disease cells, while control cultures showed only increases in fibronectin levels. Immunocytochemistry analysis also revealed extensive filamentous actin networks in disease cells, and enhanced attachment and spreading of disease cell in collagen matrices.
Three-dimensional collagen matrix cultures of primary disease cell lines are more contractile and express a more extensive filamentous actin network than patient-matched control cultures. The elevated levels of beta-catenin and Fn seen in collagen matrix cultures of disease fibroblasts can be regulated by changes in isometric tension.
杜普伊特伦挛缩症或疾病(DD)是一种手部的纤维增生性疾病,会导致特定掌腱膜束内形成瘢痕样、富含胶原蛋白的病变条索。尽管DD的分子病理学尚不清楚,但最近的证据表明β-连环蛋白可能起作用。在本研究中,原发性病变成纤维细胞的胶原基质培养显示出收缩增强以及β-连环蛋白和纤连蛋白水平的等长张力依赖性变化。
对在应激胶原基质和附着胶原基质中生长的对照和病变原代细胞培养物进行β-连环蛋白和纤连蛋白水平的蛋白质印迹分析。对胶原收缩进行定量,并对丝状肌动蛋白进行免疫细胞化学分析。
与对照细胞相比,病变细胞表现出增强的胶原收缩活性。胶原基质等长张力的改变引发了β-连环蛋白和纤连蛋白水平的显著变化,包括病变细胞内β-连环蛋白水平的短暂升高,而纤连蛋白水平稳步下降至低于正常细胞培养物中的水平。相比之下,病变细胞的附着胶原基质培养物中纤连蛋白和β-连环蛋白水平均升高,而对照培养物仅显示纤连蛋白水平升高。免疫细胞化学分析还揭示了病变细胞中广泛的丝状肌动蛋白网络,以及病变细胞在胶原基质中的附着和铺展增强。
原发性病变细胞系的三维胶原基质培养比患者匹配的对照培养更具收缩性,并且表达更广泛的丝状肌动蛋白网络。病变成纤维细胞的胶原基质培养中所见的β-连环蛋白和纤连蛋白水平升高可通过等长张力的变化来调节。