乙酰氨基酚对小鼠与人胚胎肝细胞毒性的microRNA生物标志物比较研究

Comparative study of microRNA biomarkers between human and mouse fetal liver cells after acetaminophen exposure

为探索肝损伤时miRNA的变化特点及其作为潜在生物标志物在细胞水平的跨种属特性,研究以对乙酰氨基酚(acetaminophen,APAP)为模式药物,在胚胎肝细胞上建立不同程度的细胞毒模型。采用实时荧光定量PCR的方法检测人胚胎肝细胞CCC-HEL-1和小鼠胚胎肝细胞BNL CL.2 2个种属细胞中17个miRNA在药物作用下的表达变化,并进一步分析小鼠胚胎肝细胞BNL CL.2在APAP作用不同时间后miRNA的表达变化情况。研究发现:在APAP 3个毒性剂量(IC25、IC50和IC75)作用下,17个miRNA中,超过70%的miRNA在2个种属胚胎肝细胞上表达变化趋势一致,具有跨种属的特性,特别是miR-29b在2个种属的胚肝细胞系及3个毒性剂量作用下都明显上调;在BNL CL.2细胞上,APAP作用3、6、12、24、36h后,选取的4个miRNA的表达变化具有明显时间依赖性。研究表明:miRNA作为药源性肝损伤跨种属的生物标志物,具有巨大的潜在应用价值;其中miR-29b有望成为潜在的早期指示APAP肝毒性的跨种属生物标志物。

To determine the characteristic changes of miRNA in drug-induced liver injury and their potential as the cross-species biomarkers at the cellular level,the different extents of drug-induced liver injury on the fetal liver cells were established,using the acetaminophen（APAP）as a model drug.The alterations in the expression of 17 miRNAs were detected by real-time quantitative PCR in both human（CCC-HEL-1）and mouse fetal liver cells（BNL CL.2）after APAP exposures.Moreover,time-dependent effects on the expressions of miRNAs were also determined in BNL CL.2cells after APAP treatments.The study found that more than 70% miRNAs differed in the same mammer in both human and mouse fetal liver cells after APAP exposures with the dosages of IC25,IC50,and IC75,respectively,showing the cross-species characteristic.It is worth noting that the expression levels of miR-29 bare all increased in these two cell lines after the three doses APAP treatments.Furthermore,the time-dependent effects in miRNAs expression are observed in BNL CL.2cells after 3,6,12,24,36 hAPAP exposures.The study indicates that miRNAs have the potential to be the cross-species biomarkers for the identification of drug-induced liver injury.Additionally,miR-29 bis expected to be a biomarker for the early detection of APAP-induced liver injury.