Proinflammatory chemokine induction in keratocytes and inflammatory cell infiltration into the cornea.
Proinflammatory chemokine induction in keratocytes and inflammatory cell infiltration into the cornea.
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DOI：
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发表时间：
2001-11
影响因子：
4.4
通讯作者：
J. W. Hong;J. Liu;J. S. Lee;R. R. Mohan-R.;D. Woods;Y. He;S. Wilson
中科院分区：
文献类型：
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作者：
J. W. Hong;J. Liu;J. S. Lee;R. R. Mohan-R.;D. Woods;Y. He;S. Wilson
目的探讨白细胞介素（IL）-1 α和肿瘤坏死因子（TNF）-α对角膜基质细胞中细胞因子、趋化因子和受体表达的影响，角膜刮伤对单核细胞趋化和活化因子（MCAF）表达及单核-巨噬细胞内流的影响，并探讨其在角膜基质细胞中的作用。以及MCAF和粒细胞集落刺激因子（G-CSF）微量注射对间质炎性细胞浸润的影响。方法采用基因芯片技术检测IL-1 α和TNF α对间质成纤维细胞中细胞因子、趋化因子和受体基因表达的影响。分别用RNase保护试验和Western blot分析监测MCAF mRNA和蛋白的表达。免疫细胞化学法检测兔角膜基质细胞MCAF蛋白的表达。兔角膜上皮刮伤后，用单核-巨噬细胞抗原免疫细胞化学法检测到单核-巨噬细胞。用苏木精和伊红染色监测MCAF和G-CSF微量注射到小鼠角膜基质中后的炎性细胞浸润。结果IL-1 α和TNF α可上调基质成纤维细胞中多种促炎趋化因子的表达。这些包括G-CSF、MCAF、嗜中性粒细胞活化肽（ENA-78）和单核细胞衍生的中性粒细胞趋化因子（MDNCF）。RNA酶保护试验证实MCAF mRNA表达上调，Western blot分析检测MCAF蛋白。角膜上皮损伤后4小时和24小时，在角膜基质细胞中检测到MCAF蛋白，而在未损伤角膜基质细胞中未检测到。兔角膜上皮损伤引起单核-巨噬细胞内流进入角膜基质。将MCAF和G-CSF微量注射到小鼠角膜中分别导致单核细胞-巨噬细胞和粒细胞流入基质中。结论角膜基质细胞中促炎性趋化因子的诱导是由IL-1 α和TNF α介导的。角膜细胞产生的促炎趋化因子可能在角膜手术、接触镜或创伤相关的上皮损伤后触发炎性细胞流入基质。
PURPOSE To determine the effect of interleukin (IL)-1alpha and tumor necrosis factor (TNF)-alpha on cytokine, chemokine, and receptor expression in corneal stromal cells; the effect of corneal scrape injury on monocyte chemotactic and activating factor (MCAF) expression and monocyte-macrophage influx into the stroma; and the effect of MCAF and granulocyte colony-stimulating factor (G-CSF) microinjection on inflammatory cell infiltration into the stroma. METHODS Gene array technology was used to evaluate changes in cytokine, chemokine, and receptor gene expression in stromal fibroblasts in response to IL-1alpha and TNFalpha. Expression of MCAF mRNA and protein was monitored with an RNase protection assay and Western blot analysis, respectively. Keratocyte MCAF protein expression in the rabbit cornea was detected with immunocytochemistry. After epithelial scrape injury, monocytes-macrophages were detected in rabbit corneas, by immunocytochemistry for monocyte-macrophage antigen. Inflammatory cell infiltration after MCAF and G-CSF microinjection into the stroma of mouse corneas was monitored with hematoxylin and eosin staining. RESULTS IL-1alpha or TNFalpha upregulated the expression of several proinflammatory chemokines in stromal fibroblasts in culture. These included G-CSF, MCAF, neutrophil-activating peptide (ENA-78), and monocyte-derived neutrophil chemotactic factor (MDNCF). MCAF mRNA upregulation was confirmed by RNase protection assay, and MCAF protein was detected by Western blot analysis. MCAF protein was detected in keratocytes at 4 hours and 24 hours after epithelial injury, but not in keratocytes in the unwounded cornea. Corneal epithelial injury triggered the influx of monocytes-macrophages into the corneal stroma in the rabbit. Microinjection of MCAF and G-CSF into mouse cornea resulted in the influx of monocytes-macrophages and granulocytes, respectively, into the stroma. CONCLUSIONS Proinflammatory chemokine induction in keratocytes is mediated by IL-1alpha and TNFalpha. The proinflammatory chemokines produced by the keratocytes probably trigger the influx of inflammatory cells into the stroma after epithelial injury associated with corneal surgery, contact lenses, or trauma.