Abstract: Objective To explore the relationship between the time after thrombosis and the efficacy of combined ultrasound and microbubble treatment for rescuing the ischemic tissues. Methods Rat models of thrombosis in the right common iliac artery were established and received combined ultrasound and microbubble treatment at 3, 6 and 12 h after thrombosis. The recanalization rate of the right common iliac artery was assessed using both 2-dimensional and Doppler ultrasound. The plateau acoustic intensity (AI) was quantified for estimating the skeletal microvascular blood volume, and skeletal muscle injury markers including myoglobin (Mb) and creatinine kinase (CK) were measured using ELISA. Postmortem TUNEL staining was used to detect the apoptotic rate of skeletal muscle cells in the hind limb of the rats. Results Compared with those in 3 h group, the recanalization rate and AI were significantly lower, and the levels of Mb and CK and the apoptotic rate of the skeletal muscle cells were significantly higher in both 6 h group and 12 h group (P<0.05). Compared with those in 6 h group, the rats receiving treatment at 12 h after thrombosis showed significantly lowered AI and increased Mb, CK and apoptotic rate of the skeletal muscle cells (P<0.05). Conclusion The efficacy of combined ultrasound and microbubble treatment for rescuing ischemic tissues tends to be attenuated as the time after thrombosis prolongs in rats.