【Abstract】 Objective To establish an ideal modeling method for Diarrhea Predominant Irritable Bowel Syndrome (IBS-D) with anxiety and depression in rats, and to provide an animal model for clinical study of IBS-D (IBS-D). Methods In the first stage, 20 rats were randomly divided into blank group, 3% acetic acid enema group, 4% acetic acid enema group and 5% acetic acid enema group. On the first day of intervention, the rats were given acetic acid enema with 3%, 4% and 5% concentration, respectively. After observation for 1 week, the appropriate acetic acid concentration was evaluated. In stage 2, 40 rats were randomly divided into blank group, acetic acid enema group, acetic acid + binding group, acetic acid + binding + tail clip group, and were treated with different interventions for a total of 8 days. After the intervention, the general condition, diarrhea-related index, open field test score and colonic histopathology of rats were evaluated. Results In the first stage, compared with the blank group, the fecal trait score of 4% acetic acid enema group increased at 1-3 days after intervention (P < 0.05), and gradually decreased at 4-7 days after intervention, and there was no significant difference between the group and the blank group at 7 days (P > 0.05), and the body mass was relatively lower at 1 week after intervention (P < 0.05). Fecal water content increased (P < 0.01); Compared with blank group, body mass of 5% acetic acid enema group was decreased (P < 0.05), fecal trait score, fecal water content and diarrhea index were increased (P < 0.01). There was no significant difference between 3% acetic acid enema group and blank group. Compared with the blank group in the second stage, the body mass of the simple acetic acid enema group was lower (P < 0.01), and the fecal water content and diarrhea index were increased (P < 0.01). Compared with the blank group and the simple acetic acid enema group, the body mass of the acetic acid + restraint group and the acetic acid + restraint + tail clip-on group was lower (P < 0.01). Fecal trait score, fecal water content and diarrhea index increased (P < 0.01), and colon running time decreased (P < 0.01). In the open field test score, compared with the blank group and the simple acetic acid group, the open field test distance, standing times and upright times in the acetic acid + bound group and the acetic acid + bound + clip group were all the same (P < 0.01). Compared with acetic acid + binding group, the open field experiment distance, standing times and upright times in acetic acid + binding + tail group were all different (P < 0.05). The pathological tissue of colon showed that compared with the blank group, the mucosa structure of 4% acetic acid enema group was complete with a small amount of inflammatory cell infiltration, and the pathological tissue score had no significant difference (P > 0.05), while the 5% acetic acid enema group had a medium to large amount of inflammatory cell infiltration, and the pathological tissue score was increased (P < 0.05). The mucosal structure of colon tissues in each group in stage 2 was intact, and a small amount of inflammatory cells infiltrated in different degrees. Conclusion 4% acetic acid concentration is the appropriate concentration for IBS-D modeling. After superposition and binding, IBS-D diarrhea and internal hypersensitivity characteristic state can be better simulated. After superposition of tail clip, IBS-D model of liver-stagnation and spleen-deficiency can be successfully prepared.