Objective To investigate the differentially expressed miRNAs in peripheral blood mononuclear cells (PBMCs) of ankylosing spondylitis (AS) patients, and explore its relevance with the immune inflammatory responses. Methods Fifteen AS patients (AS group) and fifteen healthy volunteers (control group) were recruited in this research. High-throughput RNA sequencing was used to screen miRNA expression in PBMCs. Real-time quantitative PCR was used to detect the six differentially expressed miRNAs. ELISA was applied to test the levels of proinflammatory cytokines, such as TNF-α, IL-1β, IL-17, and IL-23. Finally, Spearman correlation analysis was conducted to study the correlations of differentially expressed miRNAs with disease activity indicators and immune inflammatory markers. Results Forty-four miRNAs were significantly differentially expressed in AS patients, manifested as 22 up-regulated and 22 down-regulated (fold change≥1). Among them, miR-1-3p and miR-133a-5p were up-regulated obviously, while miR-127-5p, miR-345-3p and miR-136-3p were down-regulated significantly. TNF-α, IL-1β, IL-17 and IL-23 were significantly increased simultaneously in AS patients. Moreover, miR-1-3p was positively correlated with TNF-α, CRP and BASDAI score; miR-133a-5p was positively correlated with TNF-α; miR-127-5p was negatively correlated with ESR and VAS; miR-345-3p was negatively correlated with IL-17; miR-136-3p was negatively correlated with IL-17 and BASDAI score. Conclusion The miRNAs are abnormally expressed in PBMCs of AS patients, and the differentially expressed miRNAs are associated with disease activity indicators and immune inflammatory cytokines.