Atherosclerosis is the primary cause of cardiovascular and cerebrovascular diseases. Recent studies have revealed that C-X-C motif chemokine ligand 16 (CXCL16), microRNA (miR)-146a and miR-146b may have important roles in atherosclerotic diseases. However, the associations of CXCL16, miR-146a and miR-146b in atherosclerotic diseases in vivo remain unclear. Previous studies have demonstrated that miR-146a and miR-146b may negatively regulate the toll like receptor (TLR4)/nuclear factor (NF)-κB signaling pathway to repress the inflammatory response. The present study investigated the associations of CXCL16, miR-146a and miR-146b in atherosclerotic apolipoprotein E (ApoE)−/− mice in vivo. The expression levels of CXCL16, TLR4/NF-κB signaling pathway, miR-146a and miR-146b in the control and atherosclerotic ApoE−/− mice were investigated via reverse transcription-quantitative polymerase chain reaction and western blot analysis. The present study demonstrated that the expression of CXCL16 was significantly upregulated in atherosclerotic ApoE−/− mice compared with control ApoE−/− mice. The expression levels of TRL4, interleukin-1 receptor-associated kinase 1, tumor necrosis factor receptor associated factor 6, NF-κB, tumor necrosis factor-α and interleukin-1β were also significantly upregulated in atherosclerotic ApoE−/− mice compared with control mice. However, the present study revealed that the expression levels of miR-146a and miR-146b were significantly downregulated in atherosclerotic ApoE−/− mice compared with control ApoE−/− mice. Overall, the results of the present study suggested that CXCL16 may regulate the TRL4/NF-κB/CXCL16 signaling pathway, and that miR-146a and miR-146b may negatively regulate CXCL16 via this pathway in atherosclerosis in vivo. [ABSTRACT FROM AUTHOR]