Increasing evidence suggests that non-coding RNAs (ncRNA) play an important role in a variety of biological life processes by regulating gene expression at the transcriptional and post-transcriptional levels. Turbot (Scophthalmus maximus) has been threatened by various pathogens. In this study, the expression of
circular RNAs (
circRNAs),
microRNAs (
miRNAs), and
mRNA in the immune organs spleen of turbot infected with Aeromonas salmonicida was analyzed by high-throughput sequencing, and a
circRNA-
miRNA-
mRNA network was constructed, so as to explore the function of
non-coding RNA in the immune system of teleost. Illumina sequencing was performed on the uninfected group and infected group. A total of 119 differential expressed
circRNAs (DE-
circRNAs), 140 DE-
miRNAs, and 510 DE-mRNAs were identified in the four infected groups compared with the uninfected group. Most DE-mRNAs and the target genes of DE-ncRNAs were involved in immune-related pathways. The quantitative real-time PCR (qRT-PCR) results verified the reliability and accuracy of the high-throughput sequencing data. Ninety-six differentially expressed
circRNA-
miRNA-
mRNA regulatory networks were finally constructed. Among them, 15
circRNA-
miRNA-
mRNA were presented in the form of "up (
circRNA)-down (
miRNA)-up (
mRNA)" or "down-up-down". Immune-related genes gap junction CX32.2,
cell adhesion molecule 3, and
CC chemokine were also found in these networks. These results indicate that ncRNA may regulate the expression of immune-related genes through the
circRNA-
miRNA-
mRNA regulatory network and thus participate in the immune response of turbot spleen after pathogen
infection.