Interleukin-8 and monocyte chemotactic protein-1 gene expression and protein production by human orbital fibroblasts
Interleukin-8 and monocyte chemotactic protein-1 gene expression and protein production by human orbital fibroblasts
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DOI：
10.1097/00002341-199803000-00008
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发表时间：
1998-03-01
影响因子：
2
通讯作者：
Elner, SG
中科院分区：
文献类型：
--
作者：
Elner, VM;Burnstine, MA;Elner, SG
眼眶炎症很常见，但白细胞渗入眼眶组织的机制尚不清楚。我们研究了炎症过程中内毒素或重组人细胞因子对人眼眶成纤维细胞白介素8(IL-8)和单核细胞趋化蛋白-1(MCP-1)mRNA表达和蛋白分泌的影响。Northern印迹分析和ELISA法分别检测OFIL-8和MCP-1的mRNA表达和蛋白分泌。实验在不同的细胞系上进行了三次重复，重复四次。OF缺乏固有的IL-8或MCP-1基因表达，但在脂多糖或细胞因子刺激(rIL-1β和GT；rTNF-α：>；LPS和GT；干扰素-γ)作用2小时后，稳态IL-8和MCP-1 mRNA的表达显著增加，并维持在24小时。LPs)，在暴露4小时后测量(p<；0.01)。在接下来的20小时内，rIFN-γ对MCP-1的表达有明显的刺激作用(p<；0.05)，但在24 h时对IL-8的刺激作用较弱。OF分泌IL-8和MCP-1以响应内毒素和促炎细胞因子，这表明眼眶内的这些驻留细胞有能力积极参与眼眶炎症的启动和传播。旨在调节局部介质的策略可能有助于眼眶炎症性疾病的管理。
Orbital inflammation is common, but the mechanisms underlying leukocytic infiltration of orbital tissue are poorly understood. We studied resident human orbital fibroblasts (OF) interleukin-8 (IL-8), and monocyte chemotactic protein-1 (MCP-1) mRNA expression and protein secretion in response to lipopolysaccharide (LPS) or recombinant human cytokines that are present during inflammation.Third-passaged cultured human OF were left unstimulated or incubated with varying concentrations of LPS, recombinant interleukin-1-beta (rIL-1 beta), recombinant tumor necrosis factor-alpha (rTNF-alpha), or recombinant interferon-gamma (rIFN-gamma) for 2, 4, 8, or 24 h. Northern blot analysis and ELISA were performed to determine OFIL-8 and MCP-1 mRNA expression and protein secretion, respectively. Experiments were performed in triplicate and repeated four times on different cell lines.OF lacked constitutive IL-8 or MCP-1 gene expression, but produced substantial dose-dependent increases in steady-state IL-8 and MCP-1 mRNA expression by 2 h of LPS or cytokine stimulation (rIL-1 beta>rTNF-alpha: >LPS>IFN-gamma), maintained at 24 h. ELISA for IL-8 and MCP-1 proteins showed significant time-and dose-dependent OF secretion after exposure to recombinant cytokine or LPS (rIL-1 beta>rTNF-alpha>LPS), measured after 4 h of exposure (p < 0.01). This increased in the media over the next 20 h. rIFN-gamma was a potent stimulant of OF MCP-1, significant by 2 h (p < 0.05), but only a weak stimulant of IL-8 at 24 h.OF secreted IL-8 and MCP-1 in response to LPS and proinflammatory cytokines, indicating that these resident cells within the orbit have the capacity to actively participate in the initiation and propagation of orbital inflammation. Strategies aimed at modulating local mediators may be helpful in the management of orbital inflammatory disease.