Comparison of seven circulating microRNAs and ALT in carbon tetrachloride–induced liver injury model in Macaca fascicularis

• Published in: Comparative clinical pathology 2021-06, Vol.30 (3), p.379-385
• Main Authors: Yufa, Miao, Liping, Chao, Li, Sun, Guitao, Huo, Hezhan, Zhang
• Format: Article
• Language: English
• Subjects: Alanine; Alanine transaminase; Biomarkers; Carbon tetrachloride; Hematology; Liver; Macaca fascicularis; Medicine; Medicine & Public Health; MicroRNAs; miRNA; Next-generation sequencing; Oncology; Original Article; Pathology; Toxicity testing
• ISSN: 1618-5641; 1618-565X
• DOI: 10.1007/s00580-021-03228-x

Circulating microRNAs (miRNAs) are a class of noncoding RNAs and many miRNAs have been frequently reported to be potential biomarkers for drug-induced liver injury (DILI) in clinical patients. However, the usefulness of miRNAs as biomarkers for DILI in preclinical toxicological studies using non-human primates is controversial owing to the limited information. This study was initiated to determine the potential of circulating miR-122, miR-192, miR-885, miR-151, miR-378, miR-365, and miR-31 as a biomarker for carbon tetrachloride (CCl 4 )–induced liver injury in Macaca fascicularis . The ten Macaca fascicularis were orally given 15% CCl 4 (10 mL/kg) to establish the DILI. The serum at 48 h after administration when a single hepatocyte showed histopathological necrosis and vacuolation was isolated and quantified for serum miRNAs by next-generation sequencing (NGS) analysis. It was revealed that miR-122, miR-192, miR-885, miR-151, miR-378, miR-365, miR-31, and miR-802 were upregulated in the circulating serum at 48 h. The serum in different time points was collected to analyze the levels of alanine aminotransferase (ALT) and the eight miRNAs. The concentration of ALT was detected by automatic biochemistry analyzer and the miRNA level was quantified by real-time reverse-transcription PCR. All quantitative PCR values were normalized to inter-control miR-39 and calculated with the comparative Ct method. The miR-802 was not detected at each time point in the following real-time PCR amplification because of unknown reasons and would not involve any other comparison analysis in the present study. Serum ALT increased significantly from 24 h after CCl 4 -induced liver injury and reached top value at 48 h. The increasing fold changes of miR-122 were more than 8-fold from 2 h after administration and that of miR-885 were over 5-fold from 6 h. The top fold change of miR-122 was approximate 813-fold at 24 h and miR-885 was approximately 89-fold at 48 h. The miR-192 increased over a 4-fold change from 2 h and the top fold change reached approximately 119-fold at 48 h in this study. The miR-378 increased significantly at 4 h after administration and the top fold change of miR-378 was lower than the maximum fold change of ALT. The miR-365, miR-151, and miR-31 showed no correlation with serum ALT. This study indicates that serum miR-122, miR-885, and miR-192 have a stronger potential as sensitive candidate biomarkers for predicting CCl 4 -induced liver injury in Macaca fasci