Correlation of protein and gene expression profiles of inflammatory proteins after endotoxin challenge in human subjects
Correlation of protein and gene expression profiles of inflammatory proteins after endotoxin challenge in human subjects
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DOI：
10.1089/dna.2005.24.410
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发表时间：
2005-07-01
影响因子：
3.1
通讯作者：
Williams, WV
中科院分区：
文献类型：
--
作者：
Prabhakar, U;Conway, TM;Williams, WV
给人体注射内毒素(LPS)会导致深刻的生理反应，包括激活外周血单核细胞和释放炎症因子。在健康受试者(n=6)中，单次静脉注射纯化的内毒素衍生物(3.0 ng/kg)，观察其炎症反应的时程。基因芯片分析显示，一些基因的表达发生了变化，这在单独的体外内毒素刺激实验中得到了证实。随后对感兴趣基因的TaqMan分析表明，其中许多基因的表达随时间发生变化。这包括前B细胞增强因子，在微阵列分析中被鉴定为内毒素刺激后显著上调。用TaqMan分析检测基因的蛋白表达，发现在给药后1h内，血浆中开始出现肿瘤坏死因子(TNF)-α和sTNF-R蛋白，随后是其他细胞因子/炎症标志物(如IL-1ra、G-CSF、IL-6、IL-8和IL-10)和细胞因子信号抑制物(SOCS-1和SOCS-3)。总体而言，细胞因子蛋白表达与基因表达有很好的相关性；然而，一些基因表达的时间分布与蛋白质数据没有很好的相关性。对于这些蛋白质中的许多，缺乏相关性可归因于替代组织来源，这在TaqMan分析中得到了证明。主成分分析表明，细胞因子可以根据其反应的时间模式进行分组，大多数转录水平在内毒素注射后24小时恢复到基线水平。结合基因芯片和TaqMan分析来识别和量化基因表达的变化，以及蛋白质表达的分析，可以在研究炎症性疾病和其他疾病时有用。
Administration of endotoxin (LPS) in humans results in profound physiological responses, including activation of peripheral blood mononuclear cells and the release of inflammatory factors. The time course of the response of selected inflammatory proteins was examined in healthy subjects ( n = 6) administered a single intravenous dose of the purified derivative of endotoxin (3.0 ng/kg). Microarray analysis demonstrated changes in the expression of a number of genes, which were confirmed in separate in vitro endotoxin stimulation experiments. Subsequent TaqMan analysis of genes of interest indicated time-dependent changes in the expression of many of these genes. This included pre-B cell enhancing factor, which was identified on microarray analysis as being markedly upregulated following endotoxin stimulation. Protein expression of the genes examined by TaqMan analysis was measured and demonstrated the appearance of tumor necrosis factor (TNF)-alpha and sTNF-R proteins in the plasma beginning within 1 h after dosing, followed by other cytokines/ inflammatory markers ( e. g., IL-1ra, G-CSF, IL-6, IL-8, and IL-10) and suppressors of cytokine signaling ( SOCS-1 and SOCS-3). In general, cytokine protein expression correlated well with gene expression; however, the temporal profile of expression of some genes did not correlate well with the protein data. For many of these proteins, the lack of correlation was attributable to alternate tissue sources, which were demonstrated on TaqMan analysis. Principal component analysis indicated that cytokines could be grouped according to their temporal pattern of response, with most transcript levels returning to baseline 24 h following endotoxin administration. The combination of cDNA microarray and TaqMan analysis to identify and quantify changes in gene expression, along with the analysis of protein expression, can be useful in investigating inflammatory and other diseases.