Recent evidence has shown that long non-coding (lncRNA) rPvt1 is elevated in septic myocardial tissues, and knockdown of rPvt1 attenuates the sepsis induced myocardial injury. However, the mechanisms of rPvt1 in septic myocardial dysfunction have still not been illustrated. In this study, we carried out transcriptomic, proteomic and metabolomic assays, and conducted an integrated multi-omics analysis to explore the associations of rPvt1 in a lipopolysaccharide (LPS)-induced H9C2 cardiomyocyte injury. Silencing of rPvt1 was achieved via a lentivirus transduction system. Compared to the negative control, rPvt1 knockdown lead to large changes on the transcriptome, proteome and metabolome. 2385 differentially expressed genes (DEGs), 272 differentially expressed proteins (DEPs) and 75 differentially expressed metabolites (DEMs) were identified by each omics respectively. Gene Ontology (GO) functional annotation, Kyoto Encyclopedia of Genes and Genomes (KEGG), Nr, eukaryotic orthologous groups (KOG) and clusters of orthologous groups of proteins (COG) pathway analysis was conducted on these differentially expressed factors. The results suggest that mitochondrial energy metabolism may be a closely related mechanism by which Pvr1 links. These genes, proteins and metabolites and their related dysregulated pathways may become promising targets for the studies on investigating the rPvt1-regluatory mechanisms in septic myocardial dysfunction, which is important for formulating novel strategies on the prevention, diagnosis, and treatment of septic myocardial injury.