J Vet Sci.  2016 Mar;17(1):45-51. 10.4142/jvs.2016.17.1.45.

Time-course changes in the expression levels of miR-122, -155, and -21 as markers of liver cell damage, inflammation, and regeneration in acetaminophen-induced liver injury in rats

• Park HK
• Jo W
• Choi HJ
• Jang S
• Ryu JE
• Lee HJ
• Lee H
• Kim H
• Yu ES
• Son WC

• KMID: 2363334
• DOI: http://doi.org/10.4142/jvs.2016.17.1.45

Abstract

Drug-induced liver injury (DILI) is a significant threat to patient health and a major concern during drug development. Recently, multiple circulating microRNAs (miRNAs) have been reported to be potential biomarkers for DILI. To adapt and validate miRNAs for clinical use, we investigated the time-course changes in miR-122 expression levels in an acetaminophen-induced liver injury model in rats. In addition, miR-155 and miR-21 were evaluated as makers of inflammation and regeneration, respectively, to characterize liver status. Our results revealed that miR-122 is an early and sensitive biomarker of hepatocellular injury at a stage when alanine transaminase, aspartate transaminase, and total bilirubin were not detectable. However, no significant differences in the expression levels of other miRNAs (miR-155 and -21) were observed between treatment and vehicle groups. Collectively, these time-course changes in the expression levels of miRNAs may be useful as markers for clinical decision-making, in the diagnosis and treatment of DILI.

Keyword

acetaminophen; drug-induced liver injury; miR-122; miR-155; miR-21

MeSH Terms

Acetaminophen/*toxicity
Animals
Biomarkers/*blood
Chemical and Drug Induced Liver Injury/*blood/*diagnosis/pathology
Gene Expression Profiling
Gene Expression Regulation/*drug effects
Hepatocytes/*drug effects
Inflammation/blood/diagnosis
Liver Regeneration
MicroRNAs/*blood/genetics
Predictive Value of Tests
Rats
Time
Acetaminophen
Biomarkers
MicroRNAs

Figure

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  Fig. 1 Study design. Rats were treated with a single dose of acetaminophen (APAP; 1,000 mg/kg orally) or vehicle. Necropsy of animals in the vehicle group was performed 6 h after dosing, whereas animals in the treatment group were individually sacrificed at 3, 6, 24, 48, or 72 h after dosing.

• 

  Fig. 2 Histological features of liver sections. APAP-treated rats exhibited minimal to moderate centrilobular single-cell necrosis and/or apoptosis with inflammatory cell infiltration. (A) In the 6 h group, the histology of the liver was normal. (B) In the 24 h group, minimal to slight centrilobular necrosis was observed. (C) In the 48 h group, moderate to marked centrilobular necrosis with inflammation were observed. (D) In the 72 h group, slight to moderate centrilobular necrosis was observed. H&E stain. 200× (A–D). Scale bars = 50 µm.

• 

  Fig. 3 Histopathological grading of the liver. In the 3 h and 6 h groups, liver histology appeared normal. Centrilobular necrosis with inflammation was first observed in the 24 h group. The lesion grade increased in the 48 h group and decreased in the 72 h group. Error bars indicate standard error of the mean (SEM).

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  Fig. 4 Time-course changes in miR-122 expression. In the APAP-treated groups, miR-122 expression was higher than in the vehicle group. The highest fold increase was observed in the 3 h group, whereas in the 72 group, the miR-122 level was similar to that in the vehicle group. Error bars indicate SEM.

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  Fig. 5 Time-course changes in serum chemistry. Some alanine transaminase (ALT) and, aspartate transaminase (AST) levels were minimally elevated in the APAP-treated group relative to the vehicle group (ALT at 48 h; AST at 6 h); however, the magnitude of the increase was small compared to the level in the vehicle group. Total bilirubin (TBIL) levels in the APAP-treated group were not significantly elevated, and TBIL levels were not toxicologically significant. Error bars indicate SEM. *p < 0.05, **p < 0.01.

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  Fig. 6 Time-course changes in miR-155 (A) and miR-21 (B) expression levels. miR-155 and miR-21 levels were similar between treatment and vehicle (veh) groups, with no significant changes observed. Error bars indicate SEM.

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