Next Generation Sequencing Facilitates Quantitative Analysis of miRNAs in hepatic fibrosis and normal tissues.

Aim: Hepatic fibrosis is a major worldwide medical problem and can develop into liver cirrhosis and hepatocellular carcinoma(HCC). Until now, there are no effective drugs for liver ¿brosis because the molecular mechanism of progression of liver fibrosis is not fully understood. MicroRNAs (miRNAs) are an important class of small non-coding functional RNAs that play a key role in many biological processes. The purpose of this study was to clarify how the aberrant expression of miRNAs participates in development of the liver fibrosis in rat liver fibrosis model. Methods: Fibrotic and paired normal liver tissues were collected and assesssed by deep sequencing technology. MiRNA pro¿ling results were validated by quantitative real-time polymerase chain reaction (qRT-PCR) and bioinformatics was used to predict miRNA targets. Results: Nine deregulated miRNAs were induced in porcine serum (PS)-induced hepatic fibrosis versus normal liver. Further analysis revealed several signaling pathways (e.g., gap junction and neuroactive ligand-receptor interaction) may be associated with hepatic fibrogenesis. Conclusion: Several miRNAs are dysregulated in PS-induced hepatic fibrosis and seem to be closely associated with hepatic fibrogenesis. These results provide an experimental basis for understanding the mechanism of hepatic fibrosis.研究目的：肝纤维化是全球范围内的重大医学难题，可进展为肝硬化及肝细胞癌（hepatocellular carcinoma, HCC）。截至目前，尚无针对肝纤维化的有效治疗药物，其根源在于肝纤维化进展的分子机制尚未完全阐明。微小RNA（MicroRNAs, miRNAs）是一类重要的小型非编码功能性RNA，在诸多生物学过程中发挥关键调控作用。本研究旨在明确miRNAs的异常表达如何参与大鼠肝纤维化模型的肝纤维化发生发展过程。 方法：本研究收集了纤维化肝组织及配对的正常肝组织，采用深度测序技术进行检测分析。通过实时定量聚合酶链反应（quantitative real-time polymerase chain reaction, qRT-PCR）对miRNA表达谱结果进行验证，并利用生物信息学方法预测miRNA的靶基因。 结果：相较于正常肝组织，猪血清（porcine serum, PS）诱导的肝纤维化模型中存在9种差异表达的miRNAs。进一步分析显示，间隙连接、神经活性配体-受体相互作用等多条信号通路可能与肝纤维化发生相关。 结论：猪血清诱导的肝纤维化组织中存在多种异常表达的miRNAs，且其与肝纤维化发生进程紧密关联。本研究结果为阐明肝纤维化的发病机制提供了实验依据。