Osteoarthritis (OA) is an aseptic inflammatory disease which is associated with the stromal cell-derived factor 1/C-X-C chemokine receptor type 4 (SDF-1/CXCR4) axis. Accumulating studies have identified numbers of microRNAs (miRNAs) that serve important roles in the pathogenesis of OA. However, whether and how the inhibition of the SDF-1/CXCR4 axis induces alterations in miRNA expression remains largely unclear. miRNA profiling was performed in OA chondrocytes stimulated with SDF-1 alone, or SDF-1 with the CXCR4 antagonist TN14003 by miRNA microarray. Candidate miRNAs were verified by reverse transcription quantitative polymerase chain reaction. Bioinformatic analyses including target prediction, gene ontology (GO) and pathway analysis were performed to explore the potential functions of candidate miRNAs. Notably, 7 miRNAs (miR-146a-5p, miR-221-3p, miR-126-3p, miR-185-5p, miR-155-5p, miR-124-3p and miR-130a-3p) were significantly differentially expressed. GO analysis indicated that miR-146a-5p and its associated genes were enriched in receptor regulatory activity, nuclear factor-kappa-light-chain-enhancer of activated B cells (NF-B)-inducing kinase activity, cellular response to interleukin-1, cytokine-cytokine receptor interaction, NF-B signaling pathway and osteoclast differentiation pathways. CXCR4 was predicted to be a target of miR-146a-5p with high importance. The mRNA and protein levels of key factors involved in cartilage degeneration were measured following manipulation of the expression levels of miR-146a-5p in OA chondrocytes. CXCR4 and MMP-3 levels were negatively associated with miR-146a-5p expression, while the levels of type II collagen and aggrecan were positively associated. These data reveal that TN14003 upregulates miR-146a-5p expression, and also pinpoints a novel role of miR-146a-5p in inhibiting cartilage degeneration by directly targeting the SDF-1/CXCR4 axis.