Department of Physiology and Biophysics, Rosalind Franklin University of Science and Medicine, 3333 Green Bay Road, North Chicago, IL, USA,
J Cell Commun Signal. 2012 Jun;6(2):97-105. doi: 10.1007/s12079-012-0164-4. Epub 2012 May 31.
We recently show that CCN3 is a counter-regulatory molecule for the pro-fibrotic protein CCN2, and a potentially novel fibrosis therapy. The goal of this study was to assess the role of CCN3 in fibroproliferative/fibrotic responses in human dermal fibroblasts exposed to Omniscan, one of the gadolinium-based contrast agents associated with development of nephrogenic systemic fibrosis (NSF) a rare but life-threatening disease thought to be complication of NMR diagnostics in renal impaired patients. Human dermal fibroblasts were exposed to Omniscan; or to platelet-derived growth factor (PDGF) and transforming growth factor-β (TGF-β) as controls. Proliferation was assessed along with matrix metalloproteinase-1, tissue inhibitor of metalloproteinases-1 and type 1 procollagen in the absence and presence of CCN3. In parallel, CCN3 production was assessed in control and Omniscan-treated cells. The results showed that PDGF stimulated fibroblast proliferation, production of Timp-1 and MMP-1 whereas exogenous CCN3 inhibited, in a dose response manner, cell proliferation (approx. 50 % max.) and production of MMP-1 (approx 35 % max.) but had little effect on TIMP-1. TGF-β stimulated type 1 procollagen production but not proliferation, Timp-1 or MMP-1 compared to non-TGF-ß treated control cells, and CCN3 treatment blocked (approx. 80 % max.) this up-regulation. Interestingly, untreated, control fibroblasts produced high constitutive levels of CCN3 and concentrations of Omniscan that induced fibroproliferative/fibrogenic changes in dermal fibroblasts correspondingly suppressed CCN3 production. The use of PDGF and TGF-β as positive controls, and the study of differential responses, including that to Omniscan itself, provide the first evidence for a role of fibroblast-derived CCN3 as an endogenous regulator of pro-fibrotic changes, elucidating possible mechanism(s). In conclusion, these data support our hypothesis of a role for fibroblast-derived CCN3 as an endogenous regulator of pro-fibrotic changes in these cells, and suggest that CCN3 may be an important regulatory molecule in NSF and a target for treatment in this and other fibrotic diseases.
我们最近表明,CCN3 是促纤维化蛋白 CCN2 的一种反向调节分子,也是一种潜在的新型纤维化治疗方法。本研究的目的是评估 CCN3 在人真皮成纤维细胞中对促纤维增生/纤维化反应的作用,这些成纤维细胞暴露于 Omniscan 中,Omniscan 是一种与肾源性系统性纤维化(NSF)相关的钆基造影剂之一,这是一种罕见但危及生命的疾病,被认为是肾功能不全患者磁共振诊断的并发症。将人真皮成纤维细胞暴露于 Omniscan;或血小板衍生生长因子 (PDGF) 和转化生长因子-β (TGF-β) 作为对照。在不存在和存在 CCN3 的情况下,评估增殖以及基质金属蛋白酶-1、金属蛋白酶组织抑制剂-1 和 1 型前胶原。平行地,评估对照和 Omniscan 处理细胞中的 CCN3 产生。结果表明,PDGF 刺激成纤维细胞增殖、Timp-1 和 MMP-1 的产生,而外源性 CCN3 以剂量反应方式抑制细胞增殖(约 50%最大)和 MMP-1 的产生(约 35%最大),但对 TIMP-1 影响不大。与未处理的对照细胞相比,TGF-β 刺激 1 型前胶原的产生,但不刺激增殖、Timp-1 或 MMP-1,而 CCN3 处理则阻断(约 80%最大)这种上调。有趣的是,未经处理的对照成纤维细胞产生高水平的 CCN3 固有表达,并且 Omniscan 的浓度诱导真皮成纤维细胞的促纤维增生/纤维化改变,相应地抑制 CCN3 的产生。使用 PDGF 和 TGF-β 作为阳性对照,并研究包括 Omniscan 本身在内的差异反应,为成纤维细胞衍生的 CCN3 作为这些细胞中促纤维化变化的内源性调节剂提供了第一个证据,阐明了可能的机制。总之,这些数据支持我们的假说,即成纤维细胞衍生的 CCN3 作为这些细胞中促纤维化变化的内源性调节剂的作用,并表明 CCN3 可能是 NSF 中的一个重要调节分子,也是这种疾病和其他纤维化疾病治疗的一个靶点。