IL-17 stimulates MMP-1 expression in primary human cardiac fibroblasts via p38 MAPK- and ERK1/2-dependent C/EBP-β, NF-κB, and AP-1 activation
IL-17 stimulates MMP-1 expression in primary human cardiac fibroblasts via p38 MAPK- and ERK1/2-dependent C/EBP-β, NF-κB, and AP-1 activation
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DOI：
10.1152/ajpheart.00928.2007
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发表时间：
2007-12-01
影响因子：
4.8
通讯作者：
Chandrasekar, Bysani
中科院分区：
文献类型：
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作者：
Cortez, Dolores M.;Feldman, Marc D.;Chandrasekar, Bysani
基质金属蛋白酶（MMP）降解胶原并介导组织重塑。新型细胞因子IL-17在各种炎症条件下表达并调节MMP表达。我们研究了IL-17对原代人心脏成纤维细胞（HCF）MMP-1表达的影响，并描绘了相关的信号通路。用重组人IL-17处理HCF。通过北方印迹、RT-定量PCR、Western印迹和ELISA分析MMP-1表达;通过EMSA、ELISA和报告基因分析分析转录诱导和转录因子结合;通过蛋白激酶分析和Western印迹分析p38 MAPK和ERK 1/2激活。使用药理学抑制剂，小干扰RNA（siRNA）和腺病毒显性负表达载体的信号转导途径进行了研究。IL-17刺激HCF中MMP-1基因转录、净mRNA水平、蛋白质和启动子-报告基因活性。该应答被IL-17受体- Fc嵌合体和IL-17受体抗体阻断，但不被IL-6、TNF-α或IL-1 β抗体阻断。IL-17刺激的I型胶原酶活性被MMP抑制剂GM-6001和siRNA介导的MMP-1敲低抑制。IL-17刺激激活蛋白-1 [AP-1（c-Fos、c-Jun和Fra-1）]、NF-κ B（p50和p65）和CCAAT增强子结合蛋白（C/EBP）-β DNA结合和报告基因活性，反义寡核苷酸、siRNA介导的敲除或显性负性信号蛋白表达减弱了这种效应。AP-1、NF-κ B或C/EBP活化的抑制减弱了IL-17刺激的MMP-1表达。IL-17诱导p38 MAPK和ERK 1/2激活，SB-203580和PD-98059的抑制作用减弱了IL-17介导的转录因子激活和MMP-1表达。我们的数据表明，IL-17直接通过p38 MAPK和ERK依赖性AP-1、NF-κ B B和C/EBP-β激活诱导人心脏成纤维细胞中的MMP-1，并表明IL-17可能在心肌重塑中发挥关键作用。
Matrix metalloproteinases (MMPs) degrade collagen and mediate tissue remodeling. The novel cytokine IL-17 is expressed during various inflammatory conditions and modulates MMP expression. We investigated the effect of IL-17 on MMP-1 expression in primary human cardiac fibroblasts (HCF) and delineated the signaling pathways involved. HCF were treated with recombinant human IL-17. MMP-1 expression was analyzed by Northern blotting, RT-quantitative PCR, Western blotting, and ELISA; transcriptional induction and transcription factor binding by EMSA, ELISA, and reporter assay; and p38 MAPK and ERK1/2 activation by protein kinase assays and Western blotting. Signal transduction pathways were investigated using pharmacological inhibitors, small interfering RNA (siRNA), and adenoviral dominant-negative expression vectors. IL-17 stimulated MMP-1 gene transcription, net mRNA levels, protein, and promoter-reporter activity in HCF. This response was blocked by IL-17 receptor- Fc chimera and IL-17 receptor antibodies, but not by IL-6, TNF-alpha, or IL-1 beta antibodies. IL-17-stimulated type I collagenase activity was inhibited by the MMP inhibitor GM-6001 and by siRNA-mediated MMP-1 knockdown. IL-17 stimulated activator protein-1 [AP-1 (c-Fos, c-Jun, and Fra-1)], NF-kappa B (p50 and p65), and CCAAT enhancer-binding protein (C/EBP)-beta DNA binding and reporter gene activities, effects attenuated by antisense oligonucleotides, siRNA-mediated knockdown, or expression of dominant-negative signaling proteins. Inhibition of AP-1, NF-kappa B, or C/EBP activation attenuated IL-17-stimulated MMP-1 expression. IL-17 induced p38 MAPK and ERK1/2 activation, and inhibition by SB-203580 and PD-98059 blunted IL-17-mediated transcription factor activation and MMP-1 expression. Our data indicate that IL-17 induces MMP-1 in human cardiac fibroblasts directly via p38 MAPK- and ERK-dependent AP-1, NF-kappa B, and C/EBP-beta activation and suggest that IL-17 may play a critical role in myocardial remodeling.