Gene expression profiling and functional analysis of angiogenic markers in murine collagen-induced arthritis.
Gene expression profiling and functional analysis of angiogenic markers in murine collagen-induced arthritis.
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DOI：
10.1186/ar3922
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发表时间：
2012-07-20
影响因子：
4.9
通讯作者：
Paleolog EM
中科院分区：
文献类型：
--
作者：
Raatz Y;Ibrahim S;Feldmann M;Paleolog EM
血管生成失调与类风湿性关节炎（RA）的发病机制有关。为了更深入地了解关节炎相关的血管生成，我们评估了胶原诱导关节炎（CIA）发病、高峰和衰退阶段血管生成调节基因的表达，CIA是一种成熟的RA小鼠模型。用ⅱ型胶原诱导DBA/1小鼠CIA。采用活体荧光显微镜测定膝关节滑膜组织的功能毛细血管密度。为了评估关节炎关节匀浆诱导血管生成的能力，采用内皮趋化实验和体内基质塞实验。采用实时定量RT-PCR技术，采用血管生成聚焦阵列和基因特异性PCR技术，分析关节炎足血管生成相关基因的时间表达谱。最后，我们研究了特异性阻断VEGF与neuropilin (NRP)-1结合的单克隆抗体的治疗效果。尽管关节炎足匀浆在体外和体内均表现出血管生成活性，并且在组织学检查中，关节炎足滑膜表现出高度血管化，但关节炎膝滑膜的功能毛细血管密度明显降低，而毛细血管直径则明显增加。在分析的84个基因中，有41个在患关节炎的爪子中表现出与正常爪子不同的表达。最显著的改变出现在临床关节炎的高峰期。VEGF受体（Flt-1、Flk-1、Nrp-1、Nrp-2）、midkine、肝细胞生长因子、胰岛素样生长因子-1、血管生成素-1 mRNA表达增加。通过NRP-1的信号传导部分解释了在关节炎爪匀浆中观察到的内皮细胞的趋化活性。重要的是，抗nrp1b抗体的治疗性管理显著降低了CIA小鼠的疾病严重程度和进展。我们的研究结果证实，小鼠CIA关节炎滑膜是血管生成活跃的部位，但血管生成因子表达平衡的改变似乎有利于无功能和扩张的毛细血管的形成。此外，我们的研究结果证实了NRP-1在CIA发病机制中的关键作用，并支持VEGF/VEGF受体途径作为RA的潜在治疗靶点。
Dysregulated angiogenesis is implicated in the pathogenesis of rheumatoid arthritis (RA). To provide a more profound understanding of arthritis-associated angiogenesis, we evaluated the expression of angiogenesis-modulating genes at onset, peak and declining phases of collagen-induced arthritis (CIA), a well-established mouse model for RA. CIA was induced in DBA/1 mice with type II collagen. Functional capillary density in synovial tissue of knee joints was determined by intravital fluorescence microscopy. To assess the ability of arthritic joint homogenates to induce angiogenesis, an endothelial chemotaxis assay and an in vivo matrigel plug assay were employed. The temporal expression profile of angiogenesis-related genes in arthritic paws was analysed by quantitative real-time RT-PCR using an angiogenesis focused array as well as gene specific PCR. Finally, we investigated the therapeutic effect of a monoclonal antibody specifically blocking the binding of VEGF to neuropilin (NRP)-1. Although arthritic paw homogenates displayed angiogenic activity in vitro and in vivo, and synovia of arthritic paws appeared highly vascularised on histological examination, the functional capillary density in arthritic knee synovia was significantly decreased, whereas capillary diameter was increased. Of the 84 genes analysed, 41 displayed a differential expression in arthritic paws as compared to control paws. Most significant alterations were seen at the peak of clinical arthritis. Increased mRNA expression could be observed for VEGF receptors (Flt-1, Flk-1, Nrp-1, Nrp-2), as well as for midkine, hepatocyte growth factor, insulin-like growth factor-1 and angiopoietin-1. Signalling through NRP-1 accounted in part for the chemotactic activity for endothelial cells observed in arthritic paw homogenates. Importantly, therapeutic administration of anti-NRP1B antibody significantly reduced disease severity and progression in CIA mice. Our findings confirm that the arthritic synovium in murine CIA is a site of active angiogenesis, but an altered balance in the expression of angiogenic factors seems to favour the formation of non-functional and dilated capillaries. Furthermore, our results validate NRP-1 as a key player in the pathogenesis of CIA, and support the VEGF/VEGF receptor pathway as a potential therapeutic target in RA.
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影响因子：
4.4
作者：
Kim, Wan-Uk;Kang, Soon Suk;Cho, Chul-Soo
通讯作者：
Cho, Chul-Soo
影响因子：
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作者：
DeBusk, LM;Chen, Y;Lin, PC
通讯作者：
Lin, PC
影响因子：
4.4
作者：
Lu, J;Kasama, T;Adachi, M
通讯作者：
Adachi, M
影响因子：
2.9
作者：
MILLER, EJ
通讯作者：
MILLER, EJ
影响因子：
64.8
作者：
Greenberg, Joshua I.;Shields, David J.;Barillas, Samuel G.;Acevedo, Lisette M.;Murphy, Eric;Huang, Jianhua;Scheppke, Lea;Stockmann, Christian;Johnson, Randall S.;Angle, Niren;Cheresh, David A.
通讯作者：
Cheresh, David A.