Objective To analyze the changes in serum levels of macrophage inflammatory protein 3α (MIP-3α), serum amyloid A1 protein (SAA1), and glycogen synthase kinase-3β (GSK-3β) in patients with acute coronary syndrome (ACS) and their clinical significance. Methods The data of 162 ACS patients admitted to Henan Hongli Hospital from February 2023 to February 2024 were retrospectively collected and served as the research subjects. Furthermore, based on the Gensini score, patients were categorized into severe (n=50), moderate (n=46), and mild (n=66) groups according to the degree of coronary artery stenosis. Additionally, 55 healthy individuals who underwent physical examinations during the same period were selected as the healthy group. The levels of serum MIP-3α, SAA1, and GSK-3β were compared among the four groups at admission, and correlation analysis was conducted. The levels of various serum indicators were compared among patients with different prognoses at admission, and their predictive value for patient prognosis was analyzed. Results Comparison of serum MIP-3α, SAA1, and GSK-3β levels at admission among four groups: severe group > moderate group > mild group > healthy group (P<0.05); serum MIP-3α, SAA1, and GSK-3β levels at admission were positively correlated with Gensini scores (P<0.05); patients with poor prognosis had higher levels of MIP-3α, SAA1, and GSK-3β at admission than those with good prognosis (P<0.05); the AUC for predicting poor prognosis in patients based on combined detection of serum MIP-3α, SAA1, and GSK-3β levels at admission was 0.805, which was higher than that based on individual diagnosis of each indicator (P<0.05). Conclusion Patients with ACS exhibit elevated levels of serum MIP-3α, SAA1, and GSK-3β, which are associated with the severity of coronary lesions and prognosis. All three markers can serve as indicators of the severity of ACS and are of significant value in predicting patient prognosis.