Objective To evaluate the association of serum pannexin-1 （Panx-1） and stress-induced protein 2 （Sestrin2） levels with neurological severity and 30-day functional outcome in patients with acute intracerebral hemorrhage （AICH）. Methods A prospective observational cohort study was conducted involving 208 consecutive adult patients diagnosed with spontaneous AICH admitted to our center between February 2023 and April 2024. Patients were stratified by baseline National Institutes of Health Stroke Scale （NIHSS） score into mild （n = 63）， moderate （n = 94）， and severe （n = 51） neurological deficit subgroups. Two hundred age- and sex-matched healthy volunteers served as controls. Serum Panx-1 and Sestrin2 concentrations were quantified using validated ELISA kits. Group comparisons were performed using one-way ANOVA with Tukey’s post hoc test （for ≥ 3 groups） or independent-samples t-test （for two-group comparisons）. Pearson correlation coefficients assessed linear associations between biomarker levels， NIHSS score， and Glasgow Coma Scale （GCS） score. Multivariable logistic regression-adjusted for age， sex， hematoma volume （measured on baseline CT）， GCS， NIHSS， ICU length of stay， and comorbidities （cerebral herniation， pulmonary infection， deep vein thrombosis） was used to identify independent predictors of poor 30-day functional outcome （defined as modified Rankin Scale score ≥ 4）. Receiver operating characteristic （ROC） curve analysis determined the discriminatory accuracy of Panx-1， Sestrin2， and their combination for predicting poor outcome. Results Compared to control group， the levels of Panx-1 and Sestrin2 were significantly higher in AICH group （P < 0.05）. The serum levels of Panx-1 and Sestrin2 in severe defect subgroup were higher than those in mild and moderate defect subgroup （P < 0.05）. The serum levels of Panx-1 and Sestrin2 were positively correlated with NIHSS score， and negatively correlated with GCS score. Panx-1， Sestrin2， hematoma volume， GCS score， NIHSS score， ICU length of stay， concurrent cerebral herniation， concurrent pulmonary infection， and concurrent deep vein thrombosis were risk factors for poor prognosis in patients with AICH. ROC curve showed that the AUC value of combined diagnosis of Panx-1 and Sestrin2 was significantly higher than that of single diagnosis of Panx-1 and Sestrin2. Conclusions Serum Panx-1 and Sestrin2 levels are robust， severity-dependent biomarkers in AICH， independently associated with baseline neurological impairment and 30-day functional prognosis. Their synergistic integration enhances prognostic stratification accuracy beyond conventional clinical and radiological parameters， supporting their potential utility as adjunctive tools for early risk assessment and personalized therapeutic decision-making.