Inhibition of angiogenesis by interleukin-4 gene therapy in rat adjuvant-induced arthritis
Inhibition of angiogenesis by interleukin-4 gene therapy in rat adjuvant-induced arthritis
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DOI：
10.1002/art.22034
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发表时间：
2006-08-01
影响因子：
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通讯作者：
Koch, Alisa E.
中科院分区：
文献类型：
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作者：
Haas, Christian S.;Amin, M. Asif;Koch, Alisa E.
目标。白介素4(IL-4)对新生血管有调节作用。在这项研究中，我们使用基因治疗的方法来研究IL-4在大鼠佐剂性关节炎(AIA)血管生成中的作用。在关节炎发病前，大鼠关节内注射产生IL-4的腺病毒(AxCAIL-4)，这是一种没有插入的对照病毒，或对照载体(磷酸盐缓冲盐水)。在关节炎发病高峰期，大鼠被处死。测定滑膜组织中的血管形成，并评估其与炎症的相关性。在体外和体内的血管生成实验中使用踝关节匀浆，并用酶联免疫吸附试验检测细胞因子和生长因子的蛋白水平。免疫组织化学方法检测滑膜组织中αv整合素的表达。IL-4诱导滑膜组织血管密度减少，这与炎症的减少是平行的。AxCAIL-4关节匀浆在体外显著抑制内皮细胞迁移和管状形成(P<；0.05)。同样，AxCAIL-4在大鼠主动脉环实验中抑制毛细血管萌发，在体内Matrigel Plug实验中抑制血管生长。尽管有高水平的血管内皮生长因子(VEGF)，但血管抑制作用仍然存在，并且与促血管生成细胞因子IL-18、CXCL16和CXCL5的下调以及血管生成抑制物内皮抑素的上调有关。值得注意的是，AxCAIL-4还导致血管内皮细胞av和133整合素链的表达降低。在大鼠AIA中，IL-4通过抑制血管生成作用减少滑膜组织的血管生成，通过与改变的促血管生成和抗血管生成细胞因子结合来介导抑制血管生成，并可能抑制血管生成并部分通过αvβ3整合素发挥其血管抑制作用。对IL-4特定血管抑制作用的了解可能有助于优化炎症性关节炎的靶向治疗。
Objective. Interleukin-4 (IL-4) can modulate neovascularization. In this study, we used a gene therapy approach to investigate the role of IL-4 in angiogenesis in rat adjuvant-induced arthritis (AIA), a model Tor rheumatoid arthritis.Methods. Rats received an adenovirus producing IL-4 (AxCAIL-4), a control virus without insert, or control vehicle (phosphate buffered saline) intraarticularly before arthritis onset. At peak onset of arthritis, rats were killed. Vascularization was determined in the synovial tissue, and correlations with inflammation were assessed. Ankle homogenates were used in angiogenesis assays in vitro and in vivo, and protein levels of cytokines and growth factors were assessed by enzyme-linked immunosorbent assay. Synovial tissue expression of alpha v integrins was determined by immunohistochemistry.Results. IL-4 induced a reduction in synovial tissue vessel density, which was paralleled by a decrease in inflammation. AxCAIL-4 joint homogenates significantly (P < 0.05) inhibited both endothelial cell (EC) migration and tube formation in vitro. Similarly, AxCAIL-4 inhibited capillary sprouting in the rat aortic ring assay, and vessel growth in the in vivo Matrigel plug assay. The angiostatic effect occurred despite high levels of vascular endothelial growth factor (VEGF), and was associated with down-regulation of the proangiogenic cytokines IL-18, CXCL16, and CXCL5 and upregulation of the angiogenesis inhibitor endostatin. Of interest, AxCAIL-4 also resulted in decreased EC expression of the av and 133 integrin chains.Conclusion. In rat AIA, IL-4 reduces synovial tissue vascularization via angiostatic effects, mediates inhibition of angiogenesis via an association with altered pro- and antiangiogenic cytokines, and may inhibit VEGF-mediated angiogenesis and exert its angiostatic role in part via alpha v beta 3 integrin. This knowledge of the specific angiostatic effects of IL-4 may help optimize target-oriented treatment of inflammatory arthritis.