ADOA3R as a therapeutic target in experimental colitis: Proof by validated high-density oligonucleotide microarray analysis
ADOA3R as a therapeutic target in experimental colitis: Proof by validated high-density oligonucleotide microarray analysis
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DOI：
10.1097/00054725-200608000-00014
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发表时间：
2006-08-01
影响因子：
4.9
通讯作者：
Christofi, Fievos L.
中科院分区：
文献类型：
--
作者：
Guzman, Jorge;Yu, Jun Ge;Christofi, Fievos L.
腺苷A3受体（ADOA3Rs）正在成为治疗炎症性疾病的新型嘌呤能靶点。我们的目的是评估ADOA3R激动剂N(6)-（3-碘苄基）-腺苷-5-N-甲基脲胺（IB-MECA）对2,4,6-三硝基苯磺酸（TNBS）诱导的大鼠慢性结肠炎模型中基因失调和损伤的保护作用。有必要开发和验证一种微阵列技术，以测试嘌呤类药物对实验性炎症性肠病的保护作用。采用含有724个基因的RNU34神经基因芯片和SYBR绿色聚合酶链反应，对正常、tnbs处理（7天）和口服ib - meca处理（每天1.5 mg/kg）的大鼠结肠进行高密度寡核苷酸芯片分析。分析包括临床评估、体重减轻评估、电子顺磁共振成像/自旋阱监测自由基。显著的结肠炎诱导的基因失调发生在最特殊的基因群中，占基因库的5.4%，揭示了结肠炎相关的两种失调模式。下调发生在膜转运蛋白、丝裂原活化蛋白激酶和通道基因中。上调发生在趋化因子、细胞因子/炎症、应激、生长因子、细胞内信号、受体、热休克蛋白、类视黄醇代谢、神经、重塑和氧化还原敏感基因。口服IB-MECA可预防92%的这些基因失调、组织病理学、肠道损伤和体重减轻。IB-MECA或腺苷抑制体外炎症肠道中升高的自由基。口服IB-MECA阻断了结肠炎诱导的上调（90%的基因检测（37个基因中的33个））。我们的结论是，高密度寡核苷酸微阵列分析是分子基因失调研究的有力技术，可以评估嘌呤类药物或其他药物对实验性结肠炎的有益影响。ADOA3R是炎症性肠病新的潜在治疗靶点。
Adenosine A3 receptors (ADOA3Rs) are emerging as novel purinergic targets for treatment of inflammatory diseases. Our goal was to assess the protective effect of the ADOA3R agonist N(6)-(3-iodobenzyl)-adenosine-5-N-methyluronamide (IB-MECA) on gene dysregulation and injury in a rat chronic model of 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. It was necessary to develop and validate a microarray technique for testing the protective effects of purine-based drugs in experimental inflammatory bowel disease. High-density oligonucleotide microarray analysis of gene dysregulation was assessed in colons from normal, TNBS-treated (7 days), and oral IB-MECA-treated rats (1.5 mg/kg b.i.d.) using a rat RNU34 neural GeneChip of 724 genes and SYBR green polymerase chain reaction. Analysis included clinical evaluation, weight loss assessment, and electron paramagnetic resonance imaging/spin-trap monitoring of free radicals. Remarkable colitis-induced gene dysregulation occurs in the most exceptional cluster of 5.4% of the gene pool, revealing 2 modes of colitis-related dysregulation. Downregulation occurs in membrane transporter, mitogen-activated protein (MAP) kinase, and channel genes. Upregulation occurs in chemokine, cytokine/inflammatory, stress, growth factor, intracellular signaling, receptor, heat shock protein, retinoid metabolism, neural, remodeling, and redox-sensitive genes. Oral IB-MECA prevented dysregulation in 92% of these genes, histopathology, gut injury, and weight loss. IB-MECA or adenosine suppressed elevated free radicals in ex vivo inflamed gut. Oral IB-MECA blocked the colitis-induced upregulation (90% of genes tested (33 of 37 genes). We conclude that our validated high-density oligonucleotide microarray analysis is a powerful technique for molecular gene dysregulation studies to assess the beneficial effects of purine-based or other drugs in experimental colitis. ADOA3R is new potential therapeutic target for inflammatory bowel disease.