Large cerebral artery stenosis is a major cause of acute ischemic stroke (AIS); however, the correlation between serum cystatin C (CysC) and the stenosis of large cerebral arteries in patients with AIS has not been established. We performed a retrospective review of acute ischemic stroke patients, who were examined by cerebral digital subtraction angiography(DSA). Participants (252 cases) included 131 patients without stenosis and 121 patients with large cerebral artery stenosis. Serum CysC levels in patients with large cerebral artery stenosis were much higher than that of control subjects (p<0.001). However, serum CysC levels were not related to the location of stenosis. Further, logistic regression analyses showed that increased serum CysC was an independent risk factor of large cerebral artery stenosis in patients with acute ischemic stroke. Total participants were subdivided into quintiles based on serum CysC levels. Compared with the first quintile, the odds ratios of risk for large cerebral artery stenosis in the fourth and the fifth quintile were 1.26 (p<0.05) and 4.71(p<0.05) respectively, after the adjustment for age, sex, and smoking, hypertension, type 2 diabetes mellitus(DM), dyslipidemia, creatinine(Cr), urea, uric acid, and C reactive protein(CRP). Therefore, a significant positive correlation was observed between elevated serum CysC levels and large cerebral artery stenosis in patients with acute ischemic stroke. In summary, our findings provide new insights into the correlation between increased serum CysC and large cerebral artery stenosis in patients with acute ischemic stroke.