Background <p>Atherosclerosis (AS) stems from endothelial injury and inflammation. This study clarified LINC01094’s clinical implication and its role in endothelial damage and inflammation.</p> Methods <p>GEO databases screened differential lncRNAs in tissue samples from ruptured, unstable, and advanced human carotid atherosclerotic plaques. 118 patients with AS and 100 controls was enrolled, with oxidized low-density lipoprotein (ox-LDL)-stimulated Human aortic endothelial cells (HAECs) were used. Real-time quantitative reverse transcription PCR detected LINC01094 levels in serum and HAECs. Receiver operating characteristic curve analysis LINC01094’s predictive value for AS, and logistic regression identified AS risk factors. CCK-8 and flow cytometry assessed cell viability and apoptosis. Commercial kits and Enzyme-linked immunosorbent assay&#xa0;were used to quantify LDH adhesion molecules, chemokines, and inflammatory factors. DLR and RIP assays assessed miR-218-5p’s binding to LINC01094 or SP1.</p> Results <p>LINC01094 was upregulated in the GSE28829, GSE120521, and GSE21545 datasets, with notably higher expression observed in AS patients’ serum and ox-LDL-stimulated HAECs. It predicted AS occurrence with 81.36% sensitivity and 90.00% specificity, and was identified as a risk factor (OR: 8.401, 95%CI 4.175-16.907). Silencing LINC01094 countered ox-LDL-induced suppression of HAECs viability, elevation of apoptosis, LDH levels, adhesion molecules, chemokines, and inflammation. However, low miR-218-5p expression partially mitigated these effects. Overexpression of SP1 hindered the ability of miR-218-5p to alleviate ox-LDL-triggered endothelial damage and inflammation. Notably, LINC01094 positively modulated SP1 expression by sponging miR-218-5p.</p> Conclusions <p>LINC01094 is a promising biomarker for predicting AS. Moreover, silencing LINC01094 may decelerate AS progression via regulating the miR-218-5p/SP1 axis, thus alleviating endothelial injury and inflammation.</p>