Objective The effects of imiquimod on the expression of serum Tim-3 and CXCR4 in the lung tissue of the asthmatic rats were analyzed. Methods Female BALB/C rats were randomly divided into control group, model group and observation group. The control group was treated with normal saline. The model group was treated with the OVA model of asthmatic rats. The observation group was treated with imiquimod. The serum TIM-3 was detected. The SDF-1, CXCR4 and β-actin mRNA were measured. The bronchial lumen perimeter, bronchial smooth muscle area, wall area, numbers of smooth muscle cells. Results There were significantly different among bronchial wall area, bronchial smooth muscle thickness and smooth muscle nuclei, expression of SDF-1, expression of CXCR4 and TIM-3 of blank group, model group and observation group (P＜0.05). The bronchial wall area, bronchial smooth muscle thickness and smooth muscle nuclei, expression of SDF-1, expression of CXCR4 and TIM-3 of observation group were lower than that of model group (P＜0.05). Conclusion Imiquimod can significantly reduce the level of TIM-3 in serum and improve the expression of CXCR4 in lung tissue, which has a high value of application.