Effects of TGF-β2 on immune response-related gene expression profiles in the human corneal endothelium
Effects of TGF-β2 on immune response-related gene expression profiles in the human corneal endothelium
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DOI：
10.1167/iovs.03-0912
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发表时间：
2004-02-01
影响因子：
4.4
通讯作者：
Amano, S
中科院分区：
文献类型：
--
作者：
Yamagami, S;Yokoo, S;Amano, S
目的.探讨转化生长因子β 2（transforminggrowthfactor β 2，TGF β 2）对人角膜内皮细胞（humancornealendothelium，HCE）免疫应答相关基因表达谱的影响。采用人互补DNA（cDNA）表达阵列分析研究TGF-β 2对与白细胞介素（IL）-1 α和肿瘤坏死因子（TNF）-α孵育的培养HCE的影响。基因特异性半定量逆转录聚合酶链反应（RT-PCR）和酶联免疫吸附试验（ELISA）进行检查的cDNA表达阵列分析揭示的基因表达模式。此外，新鉴定的基因在HCE中的表达在人供体角膜中通过RT-PCR证实。cDNA表达阵列分析和半定量RT-PCR显示，TGF-β 2下调IL-6、生长相关（Gro）-α（CXCL 1）、单核细胞趋化蛋白（MCP）-1（CCL 2）、粒细胞集落刺激因子（G-CSF）和胰岛素样生长因子结合蛋白（IGFBP）-5的表达，上调金属蛋白酶组织抑制因子（TIMP）-1的表达。ELISA证实了TGF-β 2介导的IL-6、CXCL 1、CCL 2、G-CSF、IGFBP-5和TIMP-1在蛋白水平的表达变化。CXCL 1、G-CSF和IGFBP-5 mRNA在供体角膜HCE中首次被检测到。TGF-β 2下调IL-6、CXCL 1、CCL 2、G-CSF和IGFBP-5，并上调促炎细胞因子刺激的培养HCE中的TIMP-1，这表明TGF-β 2在房水中的免疫调节作用和TGF-β 2在眼前房中的病理生理学意义。
PURPOSE. To determine the effects of transforming growth factor (TGF)-beta2 on immune-response-related gene expression profiles in the stimulated human corneal endothelium (HCE).METHODS. A human complementary DNA (cDNA) expression array analysis was used to investigate the effects of TGF-beta2 on cultured HCE incubated with interleukin (IL)-1alpha and tumor necrosis factor (TNF)-alpha. Gene-specific semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) were performed to examine the gene expression patterns revealed by the cDNA expression array analysis. Moreover, the expression of newly identified genes in HCE was confirmed by RT-PCR in human donor corneas.RESULTS. cDNA expression array analysis and semiquantitative RT-PCR revealed that TGF-beta2 downregulated the expression of IL-6, growth-related (Gro)-alpha (CXCL1), monocyte chemotactic protein (MCP)-1 (CCL2), granulocyte-colony stimulating factor (G-CSF), and insulin-like growth factor binding protein (IGFBP)-5 and upregulated the expression of tissue inhibitor of metalloproteinase (TIMP)-1. ELISA confirmed TGF-beta2-mediated changes in the expression of IL-6, CXCL1, CCL2, G-CSF, IGFBP-5, and TIMP-1 at the protein level. CXCL1, G-CSF, and IGFBP-5 mRNAs were detected for the first time in the HCE of donor corneas.CONCLUSIONS. TGF-beta2 downregulates IL-6, CXCL1, CCL2, G-CSF, and IGFBP-5, and upregulates TIMP-1 in cultured HCE stimulated with proinflammatory cytokines, suggesting the immunomodulatory role of TGF-beta2 in the aqueous humor and the pathophysiological significance of TGF-beta2 in the anterior chamber of the eye.