Objective To investigate the role and mechanism of miRNAs in alcoholic liver injury in rats. Methods Randomly divided 30 male SD rats into model group and control group. The model group was gavaged with 56% liquor and the control group with distilled water for 8 weeks. The liver tissue was dissected. The miRNAs in the liver tissue were detected, analyzed and the target genes of differential miRNAs were predicted by rat miRNA chip. Gene Ontology (GO) and KEGG Pathway enrichment analysis were used to understand the function of differential miRNA target genes. Differential miRNA-mRNA-Pathway regulatory network was constructed using Cytoscape to further screen regulatory key miRNAs versus key pathways, qRT-PCR was performed on selected miRNAs for expression quantity validation analysis. Results: A total of 12 differentially expressed miRNAs (P < 0.05, fold change ≥ 2) were screened out, including 2 upregulated and 10 downregulated miRNAs by comparative analysis of microarray data between the model and control groups. GO classification annotation of differential miRNA target genes showed that there were close associations between differential miRNAs and biological functions such as antioxidant activity, metabolic process, biological regulation, cell killing, signal transduction, and enzyme regulatory activity. Differential miRNA target genes KEGG pathway analysis revealed that AMPK signaling pathway, PI3K Akt signaling pathway, Hippo signaling pathway, Wnt signaling pathway, cancer, autophagy, insulin resistance, Ras signaling pathway and other signaling pathways may play important regulatory roles in alcoholic liver injury lesions. The hub miRNAs and pathways screened by constructing the differential miRNA-mRNA-pathway regulatory network were mir-145-5p, mir-107-3p, mir-297, Hippo signaling pathway, cancer, PI3K Akt signaling pathway, AMPK signaling pathway, etc. The results of qRT-PCR showed that picking validation gene expression trend and gene chip results were consistent. Conclusion: In this study, we established a miRNA profile in alcoholic liver injury in rats, which suggests that mir-145-5p, mir-107-3p and mir-297 may play an important role in the process of liver alcoholic pathology.