Objective：This study aims to investigate the in vitro effects of different oligotrophic conditions on the transcriptional level of Enterococcus faecalis（E.faecalis）during starvation phase under simulated root canal pressure. Methods：The E.faecalis（ATCC 29212）was induced to starvation phase and cultured under different oligotrophic conditions（TSB with 0%，0.05%，0.15%，and 0.25% glucose），gene transcription levels and metabolic pathways were measured after one week. Results：The results showed that there was no significant change in the gene transcription level in the 0.15% glucose group compared with the 0.25% group. The gene transcription level in the 0.05% glucose group and particular in the 0% glucose group changed significantly. In the 0% glucose group，GO function mostly involved in metabolic processes and catalytic activity，and there was no significant difference in KEGG pathway. The top three metabolic pathways were ABC transporter，PTS system and amino acid biosynthesis. Conclusion：The changes in gene transcription level of E.faecalis under oligotrophic conditions may be related to the survival and pathogenicity of E.faecalis under starvation conditions. We will further explore the mechanism of related functional genes and regulatory pathways of E.faecalis in the pathogenesis of the disease on the basis of this study.