Calcium-dependent signaling in Dupuytren's disease.
Calcium-dependent signaling in Dupuytren's disease.
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DOI：
10.1007/s11552-010-9314-4
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发表时间：
2011-06-01
期刊：
影响因子：
--
通讯作者：
Levinson, Howard
中科院分区：
文献类型：
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作者：
Hadeed, Josef G;Bond, Jennifer E;Levinson, Howard
背景：以往的研究表明Dupuytren病是由成纤维细胞和肌成纤维细胞收缩引起的。平滑肌细胞中的细胞收缩性由钙依赖性和钙非依赖性信号传导机制引起。在钙依赖性途径中，钙/钙调蛋白激活肌球蛋白轻链激酶（MLCK）。在这项研究中，钙/钙调素抑制与FDA批准的药物氟奋乃静对Dupuytren的成纤维细胞的收缩性和MLCK表达的影响进行了tested.METHODS：成纤维细胞系从掌筋膜的Dupuytren病患者进行了体外研究。氟奋乃静对Dupuytren成纤维细胞迁移的影响使用划痕迁移测定法测定，收缩性使用成纤维细胞聚集的胶原晶格（FPCL）测定法测定。结果：氟奋乃静对Dupuytren成纤维细胞迁移有剂量依赖性抑制作用，在20 μ M浓度下，MLCK的抑制率最高（69.8±1.9%）。氟奋乃静也以剂量依赖性方式抑制FPCL收缩。在氟奋乃静浓度为20 μ M时观察到最大抑制（52.5±6.1%）。免疫组织化学染色表明，MLCK主要表达在整个细胞质的选择成纤维细胞内Dupuytren的结节，但不存在的Dupuytren的索和正常的掌famila.CONCLUSIONS：氟奋乃静抑制Dupuytren的成纤维细胞的收缩性和迁移，通过抑制MLCK在体外。然而，MLCK在整个Dupuytren组织中的不一致表达表明钙依赖性信号传导可能不是挛缩形成的主要模式。MLCK的氟奋乃静抑制不太可能是治疗Dupuytren病的靶点。
BACKGROUND: Previous studies suggest that Dupuytren's disease is caused by fibroblast and myofibroblast contractility. Cell contractility in smooth muscle cells is caused by calcium-dependent and calcium-independent signaling mechanisms. In the calcium-dependent pathway, calcium/calmodulin activates myosin light chain kinase (MLCK). In this study, the effects of calcium/calmodulin inhibition with the FDA-approved drug fluphenazine on Dupuytren's fibroblast contractility and MLCK expression were tested.METHODS: Fibroblast lines from the palmar fascia of patients with Dupuytren's disease were explanted and used for in vitro study. The effect of fluphenazine on Dupuytren's fibroblast migration was determined using a scratch migration assay, and contractility was determined using fibroblast-populated collagen lattice (FPCL) assays. Immunohistochemical staining of MLCK in different samples of Dupuytren's tissue and normal fascia were compared.RESULTS: Fluphenazine demonstrated a dose-dependent inhibition of Dupuytren's fibroblast migration, with the maximum inhibition of migration observed at 20muM (69.8±1.9%). Fluphenazine also inhibited FPCL contraction in a dose-dependent manner. Maximal inhibition was observed at a fluphenazine concentration of 20muM (52.5±6.1%). Immunohistological staining illustrated that MLCK was predominantly expressed throughout the cytoplasm of select fibroblasts within Dupuytren's nodules, yet was absent in the fibroblasts of Dupuytren's cords and normal palmar fascia.CONCLUSIONS: Fluphenazine inhibits Dupuytren's fibroblast contractility and migration through inhibition of MLCK in vitro. However, the inconsistent expression of MLCK throughout Dupuytren's tissue suggests that calcium-dependent signaling may not be a primary mode of contracture formation. Fluphenazine inhibition of MLCK is not likely to be a target for the treatment of Dupuytren's disease.