Identification of genes involved in drug-induced hepatotoxicity

药物引起的肝毒性相关基因的鉴定

基本信息

批准号：
16590115
负责人：
HIRANO Kazuyuki
金额：
$ 2.42万
依托单位：
Gifu Pharmaceutical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2007
项目状态：
已结题

项目摘要

In this study, we investigated the genes involved in drug-induced hepatotoxicity. The drugs we focused in this study were acetaminophen, isoniazid, metformin and buformin. To identify the genes changes their expression by the drugs in human hepatic HepG2 cells, a subtractive cDNA library was constructed. Through differential screening of the library, several clones were identified as drug induced or reduced gene. For example, acetaminophen and isoniazid increased lysyl hydroxylase, 2 (LH2) expression, and acetaminophen decreased thymidine synthetase expression. Metformin and buformin decreased glyceraldehyde 3-phosphate dehydrogenase expression and increased beta actin expression.Lysyl hydroxylase catalyzes the posttranslational hydroxylation of lysine residues in collagen. Hydroxylation of lysine residues in collagen molecules is essential for the formation and stabilization of collagen crosslinks, and is believed to act as an important role in the process of hepatic fibrosis. We therefore focused in the increased LH2 expression by the drugs. To examine the mechanism of LH2 expression increased by the drugs, we prepared the reporter plasmid containing the LH2 upstream region, and found that the increased mRNA expression was due to the transcriptional activation of LH2 by the drugs. In addition, we found that the two drugs induce the expression of transcriptional factor c-myb and the overexpression of c-myb induces LH2 mRNA expression and the transcriptional activation of LH2. From these results, increased LH2 expression may involve in the hepatotoxicity by acetaminophen and isoniazid.

在这项研究中，我们研究了参与药物诱导的肝毒性的基因。我们专注于这项研究的药物是对乙酰氨基酚，异念珠菌，二甲双胍和bugurmin。为了鉴定基因通过人肝HEPG2细胞中的药物改变其表达，构建了减去性cDNA文库。通过对文库的差异筛选，将几个克隆确定为药物诱导或降低的基因。例如，对乙酰氨基酚和异己二氮化酶增加了赖氨酸羟化酶，2（LH2）表达，对乙酰氨基酚降低了胸苷合成酶的表达。二甲双胍和bugrimin降低了3-磷酸甘油醛脱氢酶表达，并增加了β肌动蛋白表达。甘西尔羟化酶催化胶原蛋白中赖氨酸残基的翻译后羟基化。胶原分子中赖氨酸残基的羟基化对于胶原蛋白交联的形成和稳定至关重要，并且被认为在肝纤维化过程中起重要作用。因此，我们专注于药物增加的LH2表达。为了检查LH2表达的机制增加了药物的增加，我们准备了含有LH2上游区域的报告基因质粒，并发现mRNA表达增加是由于药物对LH2的转录激活所致。此外，我们发现两种药物诱导转录因子C-MYB的表达和C-MYB的过表达诱导LH2 mRNA表达和LH2的转录激活。从这些结果中，LH2表达的增加可能涉及对乙酰氨基酚和异烟肼的肝毒性。