Detecting mRNA Predictors of Acetaminophen-Induced Hepatotoxicity in Mouse Blood Using Quantitative Real-Time PCR
Detecting mRNA Predictors of Acetaminophen-Induced Hepatotoxicity in Mouse Blood Using Quantitative Real-Time PCR
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DOI：
10.1248/bpb.b15-00734
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发表时间：
2016-03-01
影响因子：
2
通讯作者：
Yomogida, Shin
中科院分区：
文献类型：
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作者：
Kanno, Syu-ichi;Tomizawa, Ayako;Yomogida, Shin
对乙酰氨基酚（APAP）是一种广泛使用的镇痛解热药。众所周知，APAP 等药物引起的肝损伤在一个物种内的个体之间存在差异。为了避免肝损伤并确保正确使用药品，能够利用遗传信息预测此类风险非常重要。本研究评估了使用定量实时聚合酶链反应 (RT-qPCR) 来识别能够预测对 APAP 诱导的肝毒性易感性的 mRNA（雄性 ddY 小鼠血液中携带）的情况。对口服 500 mg/kg APAP 的小鼠治疗后 18 小时获得的样品进行筛选。 60% 的小鼠出现 APAP 诱导的肝毒性，死亡率为 12%。有和没有 APAP 诱导的肝毒性的小鼠之间的血液 APAP 浓度没有显着差异。我们比较了 APAP 治疗组有肝毒性（阳性、严重或致命损伤）和无肝毒性小鼠之间的血液 mRNA 表达水平。在致死性损伤组中，白细胞介素编码位点 Il1β、Il10 和肿瘤坏死因子 (Tnf) 的转录水平升高。肝损伤组中编码运甲状腺素蛋白（Ttr）和金属硫蛋白1（Mtl）的基因座转录本增加，而编码谷胱甘肽过氧化物酶3（Gpx3）的基因座转录本减少。尽管禁食似乎并不影响这些基因的表达水平，但 APAP 肝毒性在禁食动物中增强。这些结果表明小鼠血液中 Il1 beta、Il10、Tnf、Ttr、Mt1 和 Gpx3 的 mRNA 表达可能提供 APAP 诱导的肝毒性的有用替代标记。
Acetaminophen (APAP) is a widely used analgesic and antipyretic drug. Drug-induced liver injury from agents such as APAP is known to vary between individuals within a species. To avoid liver injury and ensure the proper use of pharmaceutical products, it is important to be able to predict such risks using genetic information. This study evaluated the use of quantitative real-time polymerase chain reaction (RT-qPCR) to identify mRNAs (carried in the blood of male ddY mice) capable of predicting susceptibility to APAP-induced hepatotoxicity. Screening was performed on samples obtained at 18h after treatment from mice that had been orally treated with 500 mg/kg APAP. APAP-induced hepatotoxicity was seen in 60% of the mice, and the mortality rate was 12%. Blood APAP concentration did not differ significantly between mice with and without APAP-induced hepatotoxicity. We compared blood mRNA expression levels between mice with (positive, serious or lethal injury) and without hepatotoxicity in the APAP-treated group. The transcript levels of interleukin-encoding loci Il1 beta, Il10, and tumor necrosis factor (Tnf) were increased in the lethal injury group. Transcripts of the loci encoding transthyretin (Ttr) and metallothionein 1 (Mtl) showed increases in the liver injury group, while those of the glutathione peroxidase 3-encoding locus (Gpx3) were decreased. APAP hepatotoxicity was potentiated in fasted animals, although fasting did not appear to affect the level of expression of these genes. These results indicate that mRNA expression of Il1 beta, Il10, Tnf, Ttr, Mt1, and Gpx3 in mouse blood may provide useful surrogate markers of APAP-induced hepatotoxicity.