The greatest threat of arteriovenous fistula (AVF) is early thrombosis. There remains limited evidence for the use of agents for the prevention of AVF thrombosis. A total of 180 patients with stage 4 or 5 chronic kidney disease were enrolled in the present study. They were expected to have hemodialysis (HD) within the next six months and a planned lower arm AVF is expected to be the primary HD access. They were randomly divided into a control group with 60 patients, a heparin (H) treatment group with 60 patients and a heparin/anisodamine (H/A)-treatment group with 60 patients. The H/A-treatment group was given 50 IU/kg of heparin and 10 mg of anisodamine for seven days after the AVF was generated. The H-treatment group was given 50 IU/kg of heparin for seven days whereas the control group was given no treatment. The diameter and blood flow rate of the AVF were evaluated by color Doppler ultrasound at the fourth week after the operation. Patency rates of AVF were 96.7% in the H/A-treatment group, 86.7% in the H-treatment group (P < 0.05) and 83.3% in the control group (P < 0.05). The present research indicates that combined application of heparin and anisodamine can effectively relieve the vessel spasm that often occurs after establishment of an AVF and reduce the risk of early thrombosis. However, further evidence is required to validate the maintenance of long-term patency of AVF.