Neutrophil chemokines in cultured nasal broblasts
Neutrophil chemokines in cultured nasal broblasts
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DOI：
10.1034/j.1398-9995.2002.23748.x
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发表时间：
2002-12-01
期刊：
影响因子：
12.4
通讯作者：
Schroeder, JM
中科院分区：
文献类型：
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作者：
Rudack, C;Hermann, W;Schroeder, JM
背景：为了深入了解鼻窦炎中中性粒细胞迁移的机制，对原代鼻成纤维细胞合成和递送中性粒细胞趋化因子进行了分析。方法：用浓度为 20 和 200 ng/ml 的肿瘤坏死因子 (TNF)-α 处理原代鼻成纤维细胞培养物 2、8、24 和 72 小时。通过酶联免疫测定（ELISA）测定上清液中的趋化因子浓度，并通过逆转录酶聚合酶链反应（RT-PCR）测量成纤维细胞中的趋化因子mRNA表达。通过三步高效液相色谱 (HPLC) 和单博伊登室系统中测量中性粒细胞趋化性的生物测定来鉴定生物趋化活性。结果：促炎刺激在鼻成纤维细胞培养物中诱导白细胞介素 (IL)-8 和生长相关癌基因 (GRO)-α。刺激24小时后，仅检测到中性粒细胞趋化活性IL-8。刺激2小时后，粒细胞趋化蛋白（GCP）-2 mRNA已显着上调。结论：刺激后24小时和72小时，IL-8蛋白的诱导主导趋化因子的合成，而GCP-2 mRNA的诱导似乎在TNF-α暴露2小时后的早期阶段发挥作用。
Background: To gain insight into the mechanisms responsible for tissue neutrophil immigration in sinusitis, primary nasal broblasts are analyzed for synthesizing and delivering neutrophil chemokines.Methods: Primary nasal fibroblast cell culture was treated with tumor necrosis factor (TNF)-alpha concentrations of 20 and 200 ng/ml for 2, 8, 24 and 72 h. Chemokine concentrations in supernatants were determined by enzyme-linked immunoassay (ELISA) and chemokine mRNA expression in broblasts was measured by reverse transcriptase polymerase chain reaction (RT-PCR). Biological chemotactic activity was identified by three-step high-performance liquid chromatography (HPLC) and by bioassay measuring neutrophil chemotaxis in a single Boyden chamber system.Results: Interleukin (IL)-8 and growth-related oncogene (GRO)-alpha were induced in nasal fibroblast culture by proinflammatory stimulus. After 24 h of stimulation neutrophil chemotactic activity only was detected for IL-8. Granulocyte chemotactic protein (GCP)-2 mRNA was already significantly up-regulated after 2 h of stimulation.Conclusion: Induction of IL-8 protein dominates chemokine synthesis 24 and 72 h after stimulation, whereas induction of GCP-2 mRNA seems to have a role in the early phase after 2 h of exposition with TNF-alpha.