Objective The research of traditional Chinese medicine has gradually revealed a mechanism fortreating disease by regulating the microflora. The aim of this study is to establish an asthmatic mouse model with airwaymicrobial dysbiosis to provide a suitable model for studying the effects of Chinese medicine on respiratory flora and asthma.Methods Forty BALB/ c mice were randomly assigned into the control, house dust mite (HDM) asthma model, asthmamodel with intranasal administration of cefoperazone sulbactam sodium (CFP; i. n. CFP+HDM) and asthma model withintraperitoneal injection of CFP (i.p. CFP+HDM) groups. The i.n. CFP+HDM group and the i.p. CFP+HDM group wereestablished to observe the effect of antibiotics on HDM-induced asthma in mice. After 24 d of model establishment,eosinophils (EOS) in the peripheral blood ( PB) of the mice were counted. Serum IgE, TH2 cytokines in thebronchoalveolar lavage fluid (BALF) and in the lung homogenates were analyzed by ELISA. Pulmonary pathologicalchanges were observed using hematoxylin and eosin, periodic acid-Schiff and Masson staining. Results In contrast withthe control mice, EOS in the PB, serum IgE, TH2 cytokines in the BALF and lung homogenates in the HDM group weresignificantly increased ( P <0. 05), while these changes in the i. n. CFP+HDM and i. p. CFP+HDM groups were moresignificantly pronounced than those in the HDM group ( P <0. 05). Similarly, the pathological observation showed moresevere asthmatic changes in the i.n. CFP+HDM mice, but the pathology of the i.p. CFP+HDM group did not significantlydiffer from that of the model group. Conclusions Intranasally administered high-dose antibiotics, cefoperazone sulbactamsodium, aggravated HDM-induced asthma in mice, suggesting that administering antibiotics via the airway may disturb theairway microbiome and aggravate asthma occurrence. This model can be used to study Chinese medicine as a method of regulating airway microbiomes of asthma in germ-free mice.