Elastin sequences trigger transient proinflammatory responses by human dermal fibroblasts
Elastin sequences trigger transient proinflammatory responses by human dermal fibroblasts
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DOI：
10.1096/fj.13-231787
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发表时间：
2013-09-01
期刊：
影响因子：
4.8
通讯作者：
Weiss, Anthony S.
中科院分区：
文献类型：
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作者：
Almine, Jessica F.;Wise, Steven G.;Weiss, Anthony S.
皮肤穿透伤后，高度精心安排和重叠的一系列事件有助于促进伤口愈合。炎症是早期开始的一个标志，但引发和维持炎症的细胞和基质分子之间的相互关系却鲜为人知。皮肤真皮深层富含弹性蛋白。我们认为深层组织损伤包括弹性蛋白损伤，产生可溶解的弹性蛋白，引发炎症。由于真皮成纤维细胞在真皮深层占主导地位，这意味着弹性蛋白序列和成纤维细胞之间的直接相互作用将揭示促炎特征。原弹性蛋白被用作弹性蛋白序列的替代物。原弹性蛋白触发成纤维细胞表达金属弹性蛋白酶 MMP-12，该酶通常由巨噬细胞表达。 MMP-12 表达在 6 小时内增加 1056 +/- 286 倍，并持续 24 小时。趋化因子表达更加短暂，因为趋化因子C-X-C基序配体8 (CXCL8)、CXCL1和CXCL5转录物在6-12小时内分别增加11.8+/-2.6、10.2+/-0.4和8593+/-996倍，然后下降。通过使用特异性抑制剂和蛋白质截短，我们发现原弹性蛋白信号的转导是由成纤维细胞弹性蛋白结合蛋白（EBP）介导的。使用预测计算成纤维细胞模型进行的计算机模拟证实了这种上调，并且模拟显示 PKA 是信号传导回路的关键部分。我们用 1 M PKA 抑制剂 H-89 测试了这一预测，发现暴露 2 小时相应地降低了 MMP-12 (63.9 +/- 12.3%) 和所有趋化因子标记物的表达，与 EBP 抑制所见的水平一致，并验证了 PKA 作为改善促炎状态的新节点和可药物靶标。利用原弹性蛋白 C 端 RKRK 的单独触发将标记物表达降低至 65.0-76.5%，并表明整合素 (V3) 的平行参与。我们认为，真皮损伤导致的弹性蛋白溶解会导致成纤维细胞趋化因子快速上调，当暴露的弹性蛋白被 MMP-12 去除时，趋化因子就会被淬灭。Almine, J. F.、Wise, S. G.、Hiob, M.、Kumar Singh, N. K.、Tiwari, K. K.、Vali, S.、Abbasi, T. 和 Weiss, A. S. 弹性蛋白序列触发人类真皮成纤维细胞的短暂促炎反应。
Following penetrating injury of the skin, a highly orchestrated and overlapping sequence of events helps to facilitate wound resolution. Inflammation is a hallmark that is initiated early, but the reciprocal relationship between cells and matrix molecules that triggers and maintains inflammation is poorly appreciated. Elastin is enriched in the deep dermis of skin. We propose that deep tissue injury encompasses elastin damage, yielding solubilized elastin that triggers inflammation. As dermal fibroblasts dominate the deep dermis, this means that a direct interaction between elastin sequences and fibroblasts would reveal a proinflammatory signature. Tropoelastin was used as a surrogate for elastin sequences. Tropoelastin triggered fibroblast expression of the metalloelastase MMP-12, which is normally expressed by macrophages. MMP-12 expression increased 1056 +/- 286-fold by 6 h and persisted for 24 h. Chemokine expression was more transient, as chemokine C-X-C motif ligand 8 (CXCL8), CXCL1, and CXCL5 transcripts increased 11.8 +/- 2.6-, 10.2 +/- 0.4-, and 8593 +/- 996-fold, respectively, by 6-12 h and then decreased. Through the use of specific inhibitors and protein truncation, we found that transduction of the tropoelastin signal was mediated by the fibroblast elastin binding protein (EBP). In silico modeling using a predictive computational fibroblast model confirmed the up-regulation, and simulations revealed PKA as a key part of the signaling circuit. We tested this prediction with 1 M PKA inhibitor H-89 and found that 2 h of exposure correspondingly reduced expression of MMP-12 (63.9 +/- 12.3%) and all chemokine markers, consistent with the levels seen with EBP inhibition, and validated PKA as a novel node and druggable target to ameliorate the proinflammatory state. A separate trigger that utilized C-terminal RKRK of tropoelastin reduced marker expression to 65.0-76.5% and suggests the parallel involvement of integrin (V3). We propose that the solubilization of elastin as a result of dermal damage leads to rapid chemokine up-regulation by fibroblasts that is quenched when exposed elastin is removed by MMP-12.Almine, J. F., Wise, S. G., Hiob, M., Kumar Singh, N. K., Tiwari, K. K., Vali, S., Abbasi, T., and Weiss, A. S. Elastin sequences trigger transient proinflammatory responses by human dermal fibroblasts.