Objective: To observe the effect of combined application of fentanyl on stress response and postoperative pain in elderly patients undergoing cholecystectomy. Methods: Elderly patients with abdominal accepted surgery in our hospital from March 2014 to September 2015 were selected as the objects. According to the random number table, the patients were divided into control group and observation group. The control group received remifentanil anesthesia induction, the observation group were given remifentanil and sufentanil for induction of anesthesia. The changes of blood gas in perioperation, stress index and pain index were compared between the two groups. Results: After 2 h skin incision, 4 h and 12 h after surgery, the level of mean arterial pressure (MAP), heart rate (HR) in remifentanil group changed more obvious compared with observation group, and observation group recovered 12 h after surgery; the level of SpO2 in the two groups had no difference before and after anesthesia; the level of blood glucose, cortisol index in remifentanil group changed more obvious than combined treatment group, the difference was statistically different (P<0.05); the VAS scores and level of 5-HT had no significant difference between the two groups after 1 d. The VAS scores and level of 5-HT of the two groups after 3 d were lower than that of the 1 d, and the observation group decreased more significantly (P<0.05). Conclusion: Remifentanil and sufentanil induced anesthesia can reduce the stress reaction of senile perioperative patients, which can reduce the patient’s postoperative pain.