To explore the mechanism of Qingrun Formula in improving hepatic insulin resistance in rats with type 2 diabetes mellitus(T2DM) through high-throughput RNA sequencing(RNA-seq).Methods:A T2DM model of rats was established by feeding rats a high-fat diet combined with intraperitoneal injection of streptozotocin(STZ).The successfully modeled rats were randomly divided into the model group，metformin group(150 mg/kg)，and high-dose(11.2 g/kg)，medium-dose(5.6 g/kg)，and low-dose(2.8 g/kg) groups of Qingrun Formula，and a normal control group was set up.The rats were given intragastric intervention for eight weeks.Changes in fasting blood glucose(FBG)，insulin resistance index(IRI)，and insulin sensitivity index(ISI) were observed among the groups.RNA-seq technology combined with bioinformatics analysis was used to screen differentially expressed long-chain non-coding RNA(lncRNA)，micro RNA(miRNA)，and messenger RNA(mRNA).Gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG) enrichment analyses and qPCR verification of differentially expressed genes were performed，and regulatory networks of competitive endogenous RNA(ceRNA) were constructed.Results:Compared with that in the model group，the FBG in the high-dose group of Qingrun Formula decreased significantly in the 6th and 8th week of intervention(P<0.05).In the 8th week of intervention，the IRI in all groups of Qingrun Formula decreased significantly(P<0.01)，while that in the high-dose and low-dose groups of Qingrun Formula increased significantly(P<0.01，P<0.05).85 key mRNA，12 miRNA，and 37 lncRNA were obtained through differential expression screening.12 core genes were obtained by protein-protein interaction(PPI) network，and the network of lncRNA-miRNA-mRNA was constructed.KEGG enrichment analysis revealed that differentially expressed genes were mainly involved in pathways such as JAK-STAT，PPAR，amino acid metabolism，and lipid metabolism.Conclusion:Qingrun Formula may improve hepatic insulin resistance of rats with T2DM by regulating CYP2 and Acer2 genes through the lncRNA-miRNA-mRNA network and affecting Lpin1 and Insig1 genes，as well as PPAR，JAK-STAT，amino acid metabolism，lipid metabolism，and other signaling pathways.