Objective To clarify whether neurite guidance factor-1 (Netrin-1) can serve as a potential biomarker for early neurofunctional deterioration (END) risk assessment after acute ischemic stroke (AIS), providing a basis for early identification of high-risk patients in clinical practice. Methods AIS patients admitted to the Neurology Department of Mianzhu People’s Hospital between July 2022 and June 2024 with symptoms occurring within 24 hours of admission were included. At admission, the patient’s serum Netrin-1 level was measured and NIHSS scoring was performed. Within 3 days after admission, NIHSS scoring was performed 1-3 times a day to determine whether the patient had END based on the increase in NIHSS score compared to baseline, patients were categorized into END2 and END4 groups. Results A total of 384 patients were included, including 212 males, 172 females, with an average age of (63.10±9.39) years. The median level of serum Netrin-1 was 466.35 (342.85, 589.23) pg/mL. Within 72 hours after admission, 122 cases (31.8%) developed END2 and 75 cases (19.5%) developed END4. Multivariate logistic regression analysis showed that a decrease in Netrin-1 levels was an independent risk factor for END2 and END4 (P＜0.05). Conclusion The decrease in serum Netrin-1 levels is significantly correlated with an increased risk of END within 72 hours after ischemic stroke, suggesting that Netrin-1 may serve as a biological indicator for assessing END risk in patients with ischemic stroke.