Objective To explore the application effect of titanium clip hemostasis combined with hemocoagulase agkistrodon on acute non-variceal upper gastrointestinal bleeding. Methods Totally 108 patients with non-variceal upper gastrointestinal bleeding admitted to the hospital from January 2020 to July 2022 were selected as the research subjects. The patients were numbered according to the order of admission and were divided into control group (n=54, odd number) and observation group (n=54, even number) by odd-even number method. The control group was given endoscopic metal titanium clip for hemostasis, and the observation group was given endoscopic metal titanium clip combined with hemocoagulase agkistrodon for hemostasis. The hemostatic effect, improvement time of each clinical symptom and complications were compared between the two groups. The coagulation function ［fibrinogen (FBG), activated partial thromboplastin time (APTT), prothrombin time (PT), platelet (PLT)］and oxidative stress response before and after treatment and re-bleeding rate within 3 months after surgery were compared between the two groups.Results The total amount of bleeding in observation group was less than that in control group (P＜0.05), and the total effective rate of hemostasis in observation group (98.15%) was higher than that in control group (87.04%) (P＜0.05). The negative conversion time of fecal occult blood, disappearance time of hematemesis, recovery time of bowel sounds, hemostasis time, and hospital stay were significantly shorter in observation group than those in control group (P＜0.05). There were no significant differences in the incidence rates of postoperative complications between the two groups (P＞0.05). After treatment, the levels of FIB and PLT in the two groups were significantly increased (P＜0.05), and the levels in observation group were higher than those in control group (P＜0.05). APTT and PT in the two groups were significantly shortened (P＜0.05), and the two indicators in observation group were shorter compared with those in control group (P＜0.05). After treatment, the oxidative stress indicators were significantly reduced, and the indicators in observation group were lower than those in control group (P＜0.05). The re-bleeding rate within 3 months after surgery in observation group was 1.85%, which was significantly lower than 12.96% in control group (P＜0.05). Conclusion Endoscopic metal titanium clip hemostasis combined with haemocoagulase agkistrodon can enhance the hemostatic effect, shorten the remission times of clinical symptoms, optimize the oxidative stress state, improve the coagulation function, and reduce the complications and postoperative bleeding rate, and it is worthy of clinical promotion and application