Objective The incidence of coronary microvascular disease ( CMVD) is increasing annually. According to traditional Chinese medicine (TCM), CMVD belongs to the category of “collaterals”, and qi deficiency and blood stasis are the main syndrome type of CMVD. Notably however, no studies have reported on the use of animal models of CMVD with qi deficiency and blood stasis. The current study therefore aimed to establish and evaluate a rat model of CMVD with qi deficiency and blood stasis syndrome. Methods Forty-five male SD rats were divided randomly into sham group, CMVD group, and CMVD + QXXY group (n= 15 rats per group). Rats in the CMVD + QXXY group were randomly deprived of sleep for 14 ~ 16 h/ day for 6 weeks, and the model of qi deficiency syndrome was established. Animals in the sham group and the CMVD group were fed normally for 6 weeks. After 6 weeks, rats in the CMVD and CMVD + QXXY groups were anesthetized, their chests were opened, and embolic microspheres (40 ~ 120 μm) were injected into the left ventricle. Rats in the sham group underwent thoracotomy without injection of embolic microspheres. On day 7 after operation, relevant detection indicators were measured in each group. Results Compared with the sham group, the CMVD group showed a significant decrease in left ventricular ejection fraction and left ventricular shortening rate, while the activities of creatine kinase MB isoenzyme ( CK-MB) and lactate dehydrogenase ( LDH) were significantly increased. Heart function, hemorheology, myocardial enzyme index, and the degree of myocardial cell damage differed significantly between the CMVD + QXXY group compared with the sham group. Conclusions A rat model of CMVD + qi deficiency + blood stasis syndrome can be successfully established by sleep deprivation combined with intraventricular injection of embolic microspheres. This model will be suitable for the study of the pathogenesis of CMVD and the mechanisms of TCM.