Objective To investigate the protective effect of Qingyi Ⅱ on acute pancreatitis (AP) rats and its effect on ROS/JNK pathway and related inflammatory factors. Methods A total of thirty-six SD rats were randomly divided into 3 groups. Among them, 12 rats were divided into sham operation group and the other 24 SD rats were used to induce AP models. After the successful modeling, the 24 SD rats were divided into model group and intervention group according to the principle of random number table, with 12 animals in each group. Rats in the intervention group were given Qingyi Ⅱ (1ml/100 g body weight) orally, and rats in the model group were given intragastric administration of normal saline (1ml/100 g body weight). After the intervention, the pancreatic pathological score, nuclear translocation of NF-κB, tumor necrosis factor-α (TNF-α), levels of IL-6, IL-1β, reactive oxygen species (ROS), phosphorylation of JNK (p-JNK) and total JNK (JNK) were measured and compared. Results The sham operation group had lower pancreatic pathological score than the intervention group (P <0.05). The pancreatic pathological score of the intervention group was lower than that of the model group (P <0.05). Levels of TNF-α, IL-6, IL-1β, ROS, p-JNK and JNK in the model group and the intervention group were higher than those in the sham operation group (P <0.05), but the intervention group was significantly lower than the model group (P <0.05). The nuclear translocation rate of NF-kappa B in intervention group was lower than that in sham operation group and model group (P <0.05). Conclusions The pancreatic protective mechanism of Qingyi Ⅱ on AP rats may play a role in reducing the nuclear translocation level of NF-κB and down-regulating ROS/JNK pathway to reduce the expression of TNF-α, IL-6 and IL-1β.