To investigate genome-wide gene expressions of preventative moxibustion at ST25 in healthy rats. Methods:A total of 16 male Sprague-Dawley rats were randomly divided into the normal group (NC) and the normal and herb-partitioned moxibustion group (NM). Starting from day 1, the NM group received herb-partitioned moxibustion at the Tianshu points (bilateral, ST25) for 7 days. On day 8, colon tissues were collected under anesthesia with 2% pentobarbital sodium. The effect of herb-partitioned moxibustion at the ST25 in rats was observed on the general conditions, body weights, and HE staining. Three samples from each group were selected for total RNA extraction. The gene expression profile in the colon tissues was analyzed by the RNA-seq combined with bioinformatics. Results:Compared with the NC group, herb-partitioned moxibustion did not take effect on weight and histopathological observation of the colon in healthy rats. The NM group had 866 differentially expressed genes in colon tissues compared to the NC group, of which 604 were up-regulated and 262 were down-regulated. KEGG pathway analysis indicated that these genes were involved in multiple pathways, including immunity and protein metabolism. Conclusion:Herb-partitioned moxibustion at ST25 alters genome-wide gene expressions in healthy rats, and affects the biological processes and pathways. The treatment can be used as a healthcare treatment against diseases.