Laboratory of Immunology, National Eye Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
J Cell Physiol. 2012 Jan;227(1):116-26. doi: 10.1002/jcp.22708.
Chronic inflammation is implicated in the pathogenesis of age-related macular degeneration (AMD). Choroidal neovascularization (CNV) observed in exudative form of AMD results in vision loss. Human retinal pigment epithelial cell (HRPE) layer and choroidal tissue are the primary pathological sites in AMD. Pathological and therapeutic evidences have strongly indicated the vascular endothelial growth factor (VEGF) molecules as critical components in CNV pathogenesis. In these studies, we used human primary HRPE and choroidal fibroblast cells (HCHF) prepared from adult donor eyes. The effects of inflammatory cytokine (IFN-γ+ TNF-α+IL-1β) mix (ICM) on global gene expression profiles in HRPE cells, revealed 10- and 9-fold increase in VEGF-A and VEGF-C expression, respectively. The microarray results were validated by quantitative RT-PCR and secretion of VEGFs proteins. IL-1β is the most potent in inducing VEGFs secretion followed by IFN-γ and TNF-α, and the secretion was more effective in the presence of 2 and 3 cytokines. NF-κB and JAK-STAT pathway, but not HIF-1α, Sp-1, Sp-3, and STAT-3, transcription factors were upregulated and translocated to nucleus by ICM treatment. The mRNA levels of VEGF-A and VEGF-C and secretion of these proteins were also significantly enhanced by ICM in HCHF cells. The secretion of other angiogenic molecules, PEDF, SDF-1α, endostatin, and angiopoietins was not affected by ICM. Our results show that the inflammatory cytokines enhance secretion of VEGF-A and VEGF-C by HRPE and HCHF cells. These studies indicate that VEGFs secreted by these cells initiate and promote pathological choroidal and retinal noevascularization processes in AMD.
慢性炎症与年龄相关性黄斑变性（AMD）的发病机制有关。渗出性 AMD 中观察到的脉络膜新生血管（CNV）导致视力丧失。人视网膜色素上皮细胞（HRPE）层和脉络膜组织是 AMD 的主要病变部位。病理学和治疗学证据强烈表明血管内皮生长因子（VEGF）分子是 CNV 发病机制中的关键成分。在这些研究中，我们使用了从成人供体眼中制备的人原代 HRPE 和脉络膜成纤维细胞（HCHF）。炎症细胞因子（IFN-γ+TNF-α+IL-1β）混合物（ICM）对 HRPE 细胞中全局基因表达谱的影响，分别导致 VEGF-A 和 VEGF-C 的表达增加了 10 倍和 9 倍。微阵列结果通过定量 RT-PCR 和 VEGFs 蛋白的分泌进行了验证。IL-1β是诱导 VEGFs 分泌最有效的细胞因子，其次是 IFN-γ和 TNF-α，并且在存在 2 种和 3 种细胞因子时分泌效果更好。NF-κB 和 JAK-STAT 通路，但不是 HIF-1α、Sp-1、Sp-3 和 STAT-3，转录因子被 ICM 处理上调并易位到核内。ICM 还显著增强了 HCHF 细胞中 VEGF-A 和 VEGF-C 的 mRNA 水平和这些蛋白质的分泌。VEGF-A 和 VEGF-C 的分泌也显著增强了这些蛋白质的分泌。这些细胞分泌的其他血管生成分子，PEDF、SDF-1α、内皮抑素和血管生成素不受 ICM 影响。我们的结果表明，炎症细胞因子增强了 HRPE 和 HCHF 细胞中 VEGF-A 和 VEGF-C 的分泌。这些研究表明，这些细胞分泌的 VEGFs 引发并促进了 AMD 中病理性脉络膜和视网膜新生血管形成过程。
