急性肝损伤模型鼠血清中microRNAs绝对定量与肝损伤程度的相关性研究

The Correlation Between MicroRNAs in Serum and the Extent of Liver Injury

目的观察四氯化碳(CCl_4)诱导的大鼠急性肝损伤模型和对乙酰氨基酚(APAP)诱导的小鼠急性肝损伤模型血清中microRNAs(miR-122、miR-451、miR-92a、miR-192)的绝对定量随时间的变化情况,并分析血清中microRNAs绝对定量与对乙酰氨基酚诱导的小鼠急性肝损伤组织病理评分之间的相关性。方法建立CCl_4(1.5mL/kg)诱导的大鼠急性肝损伤模型和APAP(160mg/kg)诱导的小鼠急性肝损伤模型,收集建模过程中各时间点的血清,通过MiRbay^(TM) SV miRNA检测试剂盒对血清中microRNAs的含量进行绝对定量,探究血清中microRNAs的变化情况。同时收集APAP诱导的小鼠急性肝损伤模型中各时间点的肝损伤组织,进行组织病理染色、组织病理评分;随后对小鼠血清中microRNAs的绝对定量与肝损伤组织病理评分进行相关性分析。结果在两种急性肝损伤模型中,均发现血清中miR-122和miR-192的绝对定量在肝损伤8h或2h就出现明显的升高,在24h左右维持在最高的水平,72h逐渐下降至正常水平。miR-92a在CCl_4诱导的大鼠急性肝损伤模型中,出现波动性的变化,无明显的变化规律,miR-451则出现了轻微下降的趋势。miR-92a和miR-451在APAP诱导的小鼠肝损伤进展中,均有逐渐下降的趋势,在48h到达最低点,随后开始恢复。相关性分析发现,小鼠血清中miR-122和miR-192的绝对定量与APAP诱导的小鼠肝损伤组织病理评分(DILI-PSS)呈现良好的正相关(分别为r=0.741 3,P<0.05;r=0.788 3,P<0.01)。结论血清中microR-122和microR-192的绝对定量在大鼠或小鼠急性肝损伤后24h左右达峰,72h降低,并且与APAP诱导的小鼠急性肝损伤组织病理评分具有良好的相关性。

Objective To investigate the correlation between the absolute quantification of the microRNAs （miR-122, miR-451, miR-92a, miR-192） in serum during acute liver injury and the extent of liver injury on rat models of CC14 induced acute liver injury and mice models of acetaminophen （APAP） induced acute liver injury. Furthermore, to investigate the correlation between the absolute quantification of microRNAs in serum and the drug induced liver injury pathological scoring system （DILI-PSS）. Methods The acute liver injury model in rat by CCI~ （1.5 mL/kg）, and the acute liver injury model in mice by APAP （160 mg/kg） were established. The serum at different time points on both models were collected respectively. The absolute quantification of microRNAs in serum were detected by using MiRbayTM SV miRNA Assay kit. Meanwhile, the pathological sections of liver tissue of the mice at each time point were collected to analyze the correlation between microRNAs and the degree of liver injury. Results In CC14-induced rat acute liver injury model and APAP induced mouse acute liver injury, miR-122 and miR-192 appeared to be rising significantly, which remained the highest level at 24 h after treatment, and declined to the normal level after 72 h. In CC14-induced rat acute liver injury model, the change of miR-92a was fluctuated and had no apparent rules, miR-451 declined gradually, but not obviously. In mice acute liver injury model induced by APAP, miR-92a and miR-451 in the progress of liver injury declined gradually, reached the lowest point at 48 h, and then recovered. The result of correlation analysis indicated that miR-122 and miR-192 presented a good positive correlation with the DILI-PSS （r=0. 741 3,P〈0. 053 r=0. 788 3, P〈0. 01）. Conclusion The absolute quantification of miR-122 and miR-192 in serum has the highest level in 24 h, then decrease in 72 h, in both drug- induced and chemical liver injury. In addition, both the two microRNAs have good correlation with DILI-PSS in APAP-induced liver injury models.