Rokushima Masatomo, Omi Kazuo, Araki Akiko, Kyokawa Yoshimasa, Furukawa Naoko, Itoh Fumio, Imura Kae, Takeuchi Kumiko, Okada Manabu, Kato Ikuo, Ishizaki Jun
Discovery Technologies 1, Discovery Research Laboratories, Shionogi and Co Ltd, 12-4, Sagisu 5-chome, Fukushima-ku, Osaka 553-0002, Japan.
Toxicol Sci. 2007 Feb;95(2):474-84. doi: 10.1093/toxsci/kfl152. Epub 2006 Nov 2.
A variety of pharmaceutical compounds causes hemolytic anemia as a significant adverse effect and this toxicity restricts the clinical utility of these drugs. In this study, we applied microarray technology to investigate hepatic gene expression changes associated with drug-induced hemolytic anemia and to identify potential biomarker genes for this hematotoxicity. We treated female Sprague-Dawley rats with two hemolytic anemia-inducing compounds: phenylhydrazine and phenacetin. Hepatic gene expression profiles were obtained using a whole-genome oligonucleotide microarray with pooled RNA samples from individual rats within each dose group and analyzed in comparison with hepatic histopathology, hematology, and blood chemistry data. We identified a small subset of genes that were commonly deregulated in all the severe hemolytic conditions, some of which were considered to be involved in hepatic events characteristic of hemolytic anemia, such as hemoglobin biosynthesis, heme metabolism, and phagocytosis. Among them, we selected six upregulated genes as putative biomarkers, and their expression changes from microarray measurements were confirmed by quantitative real-time PCR using RNAs from individual animals. They were Alas2, beta-glo, Eraf, Hmox1, Lgals3, and Rhced. Expression patterns of all these genes showed high negative and positive correlation against erythrocyte counts and total bilirubin levels in circulation, respectively, suggesting that these genes may be the potential biomarkers for hemolytic anemia. These findings indicate that drug-induced hemolytic anemia may be detected based on hepatic changes in the expression of a subset of genes that are mechanistically linked to the hematotoxicity.
多种药物化合物会导致溶血性贫血这一严重不良反应,这种毒性限制了这些药物的临床应用。在本研究中,我们应用微阵列技术来研究与药物诱导的溶血性贫血相关的肝脏基因表达变化,并确定这种血液毒性的潜在生物标志物基因。我们用两种诱导溶血性贫血的化合物:苯肼和非那西丁处理雌性Sprague-Dawley大鼠。使用全基因组寡核苷酸微阵列从每个剂量组的个体大鼠中获取合并的RNA样本,从而获得肝脏基因表达谱,并与肝脏组织病理学、血液学和血液化学数据进行比较分析。我们确定了一小部分在所有严重溶血条件下均被共同失调的基因,其中一些基因被认为参与了溶血性贫血的肝脏相关事件,如血红蛋白生物合成、血红素代谢和吞噬作用。其中,我们选择了六个上调基因作为假定的生物标志物,通过使用个体动物的RNA进行定量实时PCR,证实了微阵列测量中它们的表达变化。它们是Alas2、β-球蛋白、Eraf、Hmox1、Lgals3和Rhced。所有这些基因的表达模式分别与循环中的红细胞计数和总胆红素水平呈现出高度的负相关和正相关,这表明这些基因可能是溶血性贫血的潜在生物标志物。这些发现表明,基于与血液毒性有机制联系的一组基因的肝脏表达变化,有可能检测出药物诱导的溶血性贫血。