Objective To establish a diarrhea rat model using multiple-stimulating factors and choosing the best indexes to assess whether the model is consistent with the disease characteristics of liver-QI stagnation with spleen deficiency in traditional Chinese medicine. Methods Newborn SD rats were randomly divided into model group (n=20) and control group (n=10). The rats of model group were stimulated by maternal separation, restraint stress and rectum acetic acid irritation, while the rats in control group were fed as normal. Weight changes, rectal sensitivity, Bristol scores and water content of feces and histology of the colon tissues were used as evaluation indexes to assess whether the model meets the demands for further studies. Results The rats in the model group showed loss of appetite, increase of water intake and urine reduction. Some rats showed increased activity, and even mania. Bristol scores and water content of feces were significantly higher than that of the control group, and the rectal sensitivity was significantly increased. The colon mucosa showed slightly thickened submucosal layer and mild inflammatory cell infiltration in the model rats. Conclusions The rat model established in this study is better to simulate the clinical manifestation of liver-QI stagnation with spleen deficiency in Chinese medicine, and may meet the demands of related researches of this disease.