C-reactive protein (CRP) emerges as an important mediator of cardiovascular lesions. In this study, we aimed to assess the role of CRP in the S-nitrosylation of proteins in endothelial cells and elucidate the potential mechanisms. Our results showed that CRP reduced protein S-nitrosylation in human umbilical vein endothelial cells (HUVECs). NO donor S-nitrosoglutathione antagonized CRP-mediated reduction of protein S-nitrosylation. Neutralizing antibody to Fcγ receptor II remarkably attenuated these changes. In addition, CRP increased NF-κB activation via the reduction of S-nitrosylation of p65, but not p50 in HUVECs, and induced the upregulation of NF-kB target gene vascular cell adhesion molecule-1. Furthermore, we confirmed that CRP reduced S-nitrosylated proteins in the rat aorta. Taken together, these data suggest that CRP-induced decline of protein S-nitrosylation by activating NF-κB via reduction of S-nitrosylation of p65, which may contribute to the endothelial dysfunction.