DNA microarray analysis of gene expression in iris and ciliary body of rat eyes with endotoxin-induced uveitis
DNA microarray analysis of gene expression in iris and ciliary body of rat eyes with endotoxin-induced uveitis
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DOI：
10.1016/j.exer.2004.10.011
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发表时间：
2005-03-01
影响因子：
3.4
通讯作者：
Yoshimura, N
中科院分区：
文献类型：
--
作者：
Ohta, K;Kikuchi, T;Yoshimura, N
本研究的目的是通过寡核苷酸微阵列系统来确定内毒素诱导的葡萄膜炎（EIU）眼睛中上调或下调的基因，并确定所选基因表达的时间和空间变化，这些基因表现出强烈的上调。通过在雄性刘易斯大鼠足垫注射脂多糖（LPS）诱导EIU。用寡核苷酸芯片分析法测定LPS注射后2、6、12和24小时虹膜睫状体（ICB）中基因的表达，并与对照组进行比较。基因芯片显示9911个基因和表达序列标签（EST）。对高度上调的基因进行聚类分析。还通过实时聚合酶链反应（PCR）研究了细胞因子（白细胞介素（IL）-1 β和IL-6）、趋化因子（RANTES）和立即早期基因（Jun B、c-Fos和c-Jun）的选定基因。免疫组化研究进行定位的一些立即早期基因产物的蛋白质表达。LPS注射后24 h，有1930个基因表达上调或下调超过2倍，其中117个基因表达上调超过10倍，这些基因可分为5个表达模式相似的基因簇。即刻早期基因和转录因子基因被包含在一个上调基因簇中，在LPS注射后2小时达到峰值。细胞因子、趋化因子和粘附分子的表达高度上调。所选基因的实时PCR分析显示出与微阵列分析检测到的相似的表达变化。注射LPS后3和6小时，在IC B细胞中发现Jun B免疫反应阳性。应用寡核苷酸芯片系统分析LPS注射后基因表达的变化。结果提示，即早基因（如Jun B）在诱导IC B中的炎症相关基因中起重要作用。(c)2004爱思唯尔有限公司保留所有权利。
The purposes of this study are to determine the genes that are up- or down-regulated in eyes with endotoxin-induced uveitis (EIU) by an oligonucleotide microarray system, and to determine the temporal and spatial changes in expression of selected genes that show strong upregulation. EIU was induced by a footpad injection of lipopolysaccharide (LPS) in male Lewis rats. The expression of genes in the iris-ciliary body (ICB) at 2, 6, 12, and 24 hr after LPS injection was determined by oligonucleotide microarray analyses and compared to that in control rats. The microarray displayed 9911 genes and expressed sequence tags (ESTs). Cluster analysis was performed for highly up-regulated genes. Selected genes for cytokines (interleukin (IL)-1 beta and IL-6), chemokines (RANTES), and immediate early genes (Jun B, c-Fos, and c-Jun) were also studied by real-time polymerase chain reaction (PCR). Immunohistochemical studies were performed to localize the protein expression of some immediate early gene products. After LPS injection, the expression of 1930 genes were increased or decreased over 2-folds compared with normal controls by 24 hr. One hundred and seventeen genes were up-regulated over 10-fold, and these were classified into five clusters with similar expression pattern. The immediate early genes and transcription factors genes were included in one cluster of up-regulated genes peaking at 2 hr after the LPS injection. The expressions of cytokines, chemokines, and adhesion molecules were highly up-regulated. Real-time PCR analyses for selected genes showed similar expression changes as detected by the microarray analyses. Jun B immunoreactivity was found in the ICB cells at 3 and 6 hr after LPS injection. Gene expression changes after LPS injection were profiled by using an oligonucleotide microarray system. Our data suggest that the immediate early genes, such as Jun B, play an important role in inducing the inflammatory-related genes in the ICB. (c) 2004 Elsevier Ltd. All rights reserved.