Integrated miRNA and mRNA Analysis Identified Potential Mechanisms and Targets of Qianggan Extracts in Preventing Nonalcoholic Steatohepatitis

Objective: Qianggan(QG) extract is a patented traditional Chinese medicine that has been widely used for the clinical treatment of nonalcoholic steatohepatitis(NASH). However, its mechanism remains unclear. Methods: The efficacy of QG was evaluated in mice with methionine-and-choline-deficient diet-induced NASH by measuring serum alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase levels and by H and E staining of liver sections. Microarray and bioinformatics analyses were performed to obtain hepatic micro RNA(mi RNA) and m RNA expression profiles and to mine potential mechanisms and therapeutic targets. Furthermore, representative mi RNA and m RNA expression levels were validated by quantitative real-time polymerase chain reaction(q RT-PCR). Results: QG extract significantly improved NASH. Twelve differentially expressed mi RNAs and 1124 differentially changed m RNAs were identified as potential targets of QG extract. Integrated analysis detected 976 mi RNA–m RNA regulatory pairs, and networks including 11 mi RNAs and 427 m RNAs were constructed by Cytoscape. Hub nodes including mi R-7050-5p, mi R-212-3p, Bcl2l11, and Kras were filtered out. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that 427 m RNAs were enriched in pathways including apoptotic process, immune response, Fox O signaling pathway, and natural killer cell-mediated cytotoxicity. We also constructed a protein–protein interaction network with 254 nodes, and identified hub genes including Kras, Fasl, and Ncam1. Finally, the results of q RT-PCR were in good accordance with microarray data. Conclusion: This study identified important hub mi RNAs and m RNAs involved in the mechanism of QG extract and which might provide potential therapeutic targets for patients with NASH.