Interleukin-17A Induction of Angiogenesis, Cell Migration, and Cytoskeletal Rearrangement
Interleukin-17A Induction of Angiogenesis, Cell Migration, and Cytoskeletal Rearrangement
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DOI：
10.1002/art.30582
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发表时间：
2011-11-01
影响因子：
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通讯作者：
Veale, Douglas J.
中科院分区：
文献类型：
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作者：
Moran, Ellen M.;Connolly, Mary;Veale, Douglas J.
目标。研究白细胞介素- 17a （IL-17A）刺激血管生成、细胞迁移和细胞骨架重排的能力。采用Matrigel基质和Transwell Matrigel侵袭室评价IL-17A对人真皮内皮细胞（HDECs）微血管管形成和细胞外基质侵袭的影响。采用酶联免疫吸附法测定il - 17a诱导的类风湿性关节炎滑膜成纤维细胞（RASFs）和HDECs中生长相关癌基因α (GRO α)和单核细胞趋化蛋白1 （MCP-1）的产生。使用或不使用抗gro α和抗mcp -1抗体，通过外周血单个核细胞（PBMC）迁移试验和伤口修复划痕试验评估il - 17a诱导的迁移。使用整合素结合试验评估β 1整合素受体的结合。通过f -肌动蛋白免疫荧光染色评估rasf和HDECs的细胞骨架组装/拆卸情况。在Rac1抑制剂（NSC23766）存在的情况下，评估了il - 17a诱导的细胞迁移和细胞骨架分解。通过Rac GTPase下拉法和Western blot检测IL-17刺激后，抗gro α、抗mcp -1或IgG对照存在或不存在时，Rac1的激活情况。IL-17A显著上调血管生成和内皮细胞侵袭。显著诱导rasf中GRO α和MCP-1的表达。IL-17A诱导PBMCs、rasf和HDECs迁移；这些作用被抗gro α或抗mcp -1抗体阻断。IL-17A在rasf和HDECs中显著上调β 1整合素受体结合并诱导细胞骨架解体。IL-17A直接诱导Rac1激活。il - 17a诱导的伤口修复和肌动蛋白重排被Rac1药理学抑制剂（NSC23766）抑制。抗gro α或抗mcp -1抗体对il - 17a诱导的Rac1激活无影响。IL-17A诱导血管生成、细胞迁移和细胞侵袭，所有这些都是类风湿关节炎发病的关键过程，部分是通过趋化因子和细胞骨架依赖途径介导的。
Objective. To examine the ability of interleukin-17A (IL-17A) to stimulate angiogenesis, cell migration, and cytoskeletal rearrangement.Methods. The effect of IL-17A on microvascular tube formation and extracellular matrix invasion by human dermal endothelial cells (HDECs) was assessed using Matrigel matrix and Transwell Matrigel invasion chambers. IL-17A-induced growth-related oncogene alpha (GRO alpha) and monocyte chemotactic protein 1 (MCP-1) production in rheumatoid arthritis synovial fibroblasts (RASFs) and HDECs was measured by enzyme-linked immunosorbent assay. IL-17A-induced migration was assessed using peripheral blood mononuclear cell (PBMC) migration assays and wound-repair scratch assays, with or without anti-GRO alpha and anti-MCP-1 antibodies. Binding of beta 1 integrin receptors was assessed using integrin binding assays. Cytoskeletal assembly/disassembly in RASFs and HDECs were assessed by immunofluorescence staining for F-actin. IL-17A-induced cell migration and cytoskeletal disassembly were assessed in the presence of a Rac1 inhibitor (NSC23766). Rac1 activation following IL-17 stimulation in the presence or absence of anti-GRO alpha, anti-MCP-1, or IgG control was assessed by Rac GTPase pull-down assays and Western blotting.Results. IL-17A significantly up-regulated angiogenesis and endothelial cell invasion. It significantly induced GRO alpha and MCP-1 expression in RASFs. Migration of PBMCs, RASFs, and HDECs was induced by IL-17A; these effects were blocked by anti-GRO alpha or anti-MCP-1 antibodies. IL-17A significantly up-regulated beta 1 integrin receptor binding and induced cytoskeletal disassembly in RASFs and HDECs. Rac1 activation was directly induced by IL-17A. IL-17A-induced wound repair and actin rearrangement were inhibited by a pharmacologic inhibitor of Rac1 (NSC23766). Anti-GRO alpha or anti-MCP-1 antibodies had no effect on IL-17A-induced Rac1 activation.Conclusion. IL-17A induces angiogenesis, cell migration, and cell invasion, all of which are key processes in the pathogenesis of rheumatoid arthritis and ones that are mediated in part through chemokine-and cytoskeleton-dependent pathways.