Suzuki Hiromi, Inoue Tomoaki, Matsushita Tomochika, Kobayashi Kazuko, Horii Ikuo, Hirabayashi Yoko, Inoue Tohru
Fuji-Gotemba Research Laboratories, Chugai Pharmaceutical Co. Ltd, 1-135 Komakado, Gotemba, Shizuoka 412-8513, Japan.
J Appl Toxicol. 2008 Mar;28(2):227-36. doi: 10.1002/jat.1328.
The study examined the feasibility of screening for hepatotoxicity by an in vitro gene expression analysis using rat primary hepatocytes and Affymetrix Rat Toxicology U34 arrays. Hepatocytes were exposed for 6 or 24 h to eight drugs, with different mechanisms of hepatotoxicity, at one third of the cytotoxic concentration TC50, i.e. acetaminophen, cyclophosphamide, clofibrate, chlorpromazine, lithocholic acid, cisplatin, diclofenac and disulfiram. The types of transcriptional changes observed in this study were generally consistent with previously reported in vivo data, although there were some differences. In hierarchical cluster analysis, drugs formed clusters depending on their mode of toxicity against cells. The number of transcripts affected by the cholestatic hepatotoxicants (lithocholic acid and chlorpromazine) or the drugs that rarely cause of hepatotoxicity (cisplatin, diclofenac and disulfiram) were limited compared with the other drugs (acetaminophen, clobifibrate and cyclophosphamide), where they did not induce transcriptional changes apparently related to toxicity. It is concluded that in vitro gene expression analysis of hepatocytes using microarray is a useful tool for evaluating the toxicological profile of drugs and in screening for the direct toxicity of drugs against hepatocytes.
该研究使用大鼠原代肝细胞和Affymetrix大鼠毒理学U34芯片,通过体外基因表达分析来检测肝毒性筛查的可行性。将肝细胞暴露于具有不同肝毒性机制的8种药物中6或24小时,浓度为细胞毒性浓度TC50的三分之一,即对乙酰氨基酚、环磷酰胺、氯贝丁酯、氯丙嗪、石胆酸、顺铂、双氯芬酸和双硫仑。尽管存在一些差异,但本研究中观察到的转录变化类型总体上与先前报道的体内数据一致。在层次聚类分析中,药物根据其对细胞的毒性模式形成簇。与其他药物(对乙酰氨基酚、氯贝丁酯和环磷酰胺)相比,胆汁淤积性肝毒性药物(石胆酸和氯丙嗪)或很少引起肝毒性的药物(顺铂、双氯芬酸和双硫仑)影响的转录本数量有限,在这些药物中它们并未诱导明显与毒性相关的转录变化。结论是,使用微阵列对肝细胞进行体外基因表达分析是评估药物毒理学特征和筛查药物对肝细胞直接毒性的有用工具。