Objective To investigate the role and mechanism of miRNAs in alcoholic liver injury in rats. Methods Thirty male SD rats were randomly divided into model and control groups. The model group was gavaged with 56% liquor and the control group was gavaged with distilled water for 8 weeks. Liver tissue was collected, miRNAs were analyzed, and target genes of differentially expressed miRNAs were predicted by a rat miRNA chip. Gene ontology (GO) and KEGG pathway enrichment analysis were used to understand the function of differentially expressed miRNA target genes. A differentially expressed miRNA-mRNA-pathway regulatory network was constructed using Cytoscape to further screen important regulatory miRNAs versus important pathways. RT-qPCR was performed for selected miRNAs to validate the expression analysis. Results Twelve differentially expressed miRNAs (P<0. 05, Fold change≥2) were screened out, including two upregulated and 10 downregulated miRNAs by comparative analysis of microarray data between model and control groups. GO classification annotation of differential miRNA target genes showed close associations between differentially expressed miRNAs and biological functions such as signal transduction, metabolic processes, antioxidant activity, cell killing, enzyme regulatory activity and biological regulation. Differentially expressed miRNA target genes in KEGG pathway analysis revealed that the AMPK signaling pathway, PI3K-Akt signaling pathway, Hippo signaling pathway, Wnt signaling pathway, cancer, autophagy, insulin resistance, Ras signaling pathway, and other signaling pathways might play major regulatory roles in alcoholic liver injury lesions. Hub miRNAs and pathways screened by constructing the differentially expressed miRNA-mRNA-pathway regulatory network were miR-145-5p, miR-107-3p, miR-297, Hippo signaling pathway, cancer, PI3K-Akt signaling pathway, and AMPK signaling pathway. qRT-PCR validated the gene expression trends, and gene chip result were consistent. Conclusions We established an miRNA profile of alcoholic liver injury in rats, which suggests that miR-145-5p, miR-107-3p, and miR-297 play major roles in the process of alcoholic liver pathology.