Objective To investigate the possibility of establishing the high?cholesterol forage?feeding tree shrew as a cholesterol?induced gallstone model and to determine the association of substance P (SP) with this model. Methods Upon establishment of the tree shrew cholesterol?induced gallstone model, the SP expression of the digestive tract was detected with immunohistochemistry method and compared among the model groups and the control group. Results In the model groups, immunohistochemistry showed that gastrointestinal SP expression was lower than that in the control group ( P < 0. 05). The tree shrew gallstone model was negatively correlated with SP expression level. Conclusions Tree shrew gallstones are associated with the regulation mechanisms of hormone secretion in the digestive tract.