Silica nanoparticles (SiO NPs) are extensively applied in various field, which increased their health risks to humans. SiO NPs were reported to enter into blood through inhalation and meanwhile, the potential use of SiO NPs as drug carriers in vivo allows them to present in blood circulation to induce inflammation of vascular endothelial cells which can be closely related with cardiovascular diseases, whilst the intrinsic mechanism has not been well understood. In this study, we found a regulation of signal axis induced by amorphous SiO NPs that triggers pro-inflammatory responses in human umbilical vein endothelial cells (HUVECs). HUVECs exposed with SiO NPs generate excess amount of reactive oxygen species (ROS) and lactate dehydrogenase (LDH), together with the up-regulation of cell inflammatory factors [interleukin-1 beta (IL-1β), interleukin-6 (IL-6), tumor necrotic factor-α (TNF-α)] and cell adhesion molecules [intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1)]. In addition, SiO NPs were found to promote the translocation and release of high-mobility group box 1 (HMGB1) from nucleus to cytoplasm, which was demonstrated to be regulated by ROS and NOD-like receptor pyrin domain containing 3 (NLRP3) inflammasome. Subsequently, toll-like receptor 4 (TLR4) could bind with HMGB1, up-regulate the expression of myeloid differentiation factor 88 (MyD88) and then activate nuclear factor kappa-B (NF-κB) signaling pathway, ultimately induced the inflammatory response of HUVECs. Overall, out results revealed the related signaling pathways of cell inflammation induced by amorphous SiO NPs, which provided new insights in understanding SiO NPs-induced cytotoxicity and offered safety guidance for further nanomaterial application.