Objective To explore the establishment and evaluation of a rat model of coronary microvascular disease with Qi deficiency and blood stasis syndrome. Methods 45 male SD rats were randomly divided into sham operation group, coronary microvascular disease group and coronary microvascular qi deficiency and blood stasis syndrome group, with 15 rats in each group. The rats in the group of coronary microvascular disease with Qi deficiency and blood stasis syndrome were randomly deprived of sleep for 14 to 16 hours every day for 6 weeks, and the model of Qi deficiency syndrome was established. The animals in the sham operation group and the coronary microvascular disease group were fed normally for 6 weeks. Six weeks after the experiment, rats in the coronary microvascular disease group and coronary microvascular qi deficiency and blood stasis syndrome group were anesthetized, and then opened their chest and injected embolic microspheres (40~120μm) into the left ventricle,The animals in the sham operation group underwent thoracotomy without injection of embolic microspheres. On the 7th day after operation, relevant detection indicators were measured in each group. Results In the group of coronary microvascular disease, microsphere embolism was found in coronary microvascular, left ventricular ejection fraction and left ventricular shortening rate were significantly decreased, and the activities of creatine kinase MB isoenzyme (CK-MB) and lactate dehydrogenase (LDH) were significantly increased. The heart function, hemorheology, myocardial enzyme index and the degree of myocardial cell damage in the coronary microvascular Qi deficiency and blood stasis syndrome group were significantly different from those in the sham operation group. Conclusion Sleep deprivation combined with intraventricular injection of embolic microsphere can successfully establish a rat model of coronary microvascular disease with qi deficiency and blood stasis syndrome, which is more suitable for the study of pathogenesis and mechanism of traditional Chinese medicine