Abstract: Objective To study the effect of anisodamine combined with neostigmine on inflammatory response in rat after bilioenteric anastomosis. Methods A total of 30 SD rats were randomly divided into 5 groups, 6 in each group, including normal group, control group, low dose group, normal dose group and high dose group. Rats in normal group were sacrificed right now after grouping. Rats in normal dose group received anisodamine (25 mg/kg) combined with neostigmine (50μg/kg) on the first day after operation by intraperitoneal injection, and continued for 7 days. Low dose group rats received half of the normal dose (anisodamine for 12.5 mg/kg and neostigmine for 25μg/kg), high dose group rats received 2 times of the normal dose (anisodamine for 50 mg/ kg and neostigmine for 100μg/kg), and rats in control group were given normal saline. The weight and inflammation indexes (CRP, TNF-α, IL-1β) of rats in different groups were compared at 1 week after operation. Results The operation was successfully completed in each rat. No rats died except one in the high dose group after the second day of injection. One week after operation, TNF-α, IL-1β of rats in control group, low dose group, normal dose group and high dose group were significantly higher than normal group (P＜ 0.05). While, there were no significant differences in weight and inflammation indexes of rats in low dose group and control group. The results of CRP, TNF-α and IL-1β of normal dose group were (2.7±0.1) mg/L, (1.8±0.2) pg/ml and (69.5±6.3) pg/ml, which were significantly lower than those of control group (P＜ 0.05). Conclusion Anisodamine combined with neostigmine can effectively reduce the systemic inflammatory response in rats after bilioenteric anastomosis. The optimal drug concentration is anisodamine (25 mg/kg) combined with neostigmine (50μg/kg), the drug effect of low dose is not significant, and high dose of the drugs may have a risk of causing the death of animals.