Increased high-mobility group box 1 (HMGB1) levels were found in patients after acute ischemic stroke. The aim of this study was to examine whether the circulating HMGB1 levels could predict the 3-month post-stroke depression (PSD). The subjects were first-ever ischemic stroke patients who were hospitalized during the period from July 2020 to December 2020. HMGB1 concentrations were measured by enzyme-linked immunosorbent assay after admission. A 24-item Hamilton Depression Rating Scale was performed to evaluate PSD at 3months after stroke. The analyses included 324 participants (mean age, 63.7years; 171 male). Ninety-four patients (29.0%) were diagnosed as having PSD at 3months. The median serum HMGB1 levels at admission was 7.5ng/mL (IQR, 4.4-11.3ng/mL). The PSD distribution across the HMGB1 quartiles ranged between 17.5% (first quartile) and 57.5% (fourth quartile). After covariate adjustments, the fourth quartile of HMGB1 was found to be associated with a higher risk of PSD (as compared with first HMGB1 quartile, odd ratio, 1.26; 95% confidence interval [CI], 1.17-1.35; P < 0.001). The area under the receiver operating characteristic curve of HMGB1 was 0.726 (95% CI 0.660-0.792) for PSD. Similar results were found when HMGB1 was analyzed as continuous variable. Furthermore, the optimal cutoff point of circulating HMGB1 levels was 8.6ng/mL, with a sensitivity of 69.2% and a specificity of 73.9%. This study demonstrated that higher HMGB1 levels in the acute phase of ischemic stroke were associated with increased risk of PSD.