Aim To observe the expression of miR-221-3p in ischemic disease of lower extremity. MethodsUsing microarray to screen the different expression of miR-221-3p in arteriosclerosis obliterans(ASO) patients, then real time-PCR was further used to confirm the changes in 55 ASO patients and 54 healthy volunteers. Male SD rats were divided into hind-limb ischemia groups, sham operation group respectively. The laser Doppler imager was performed to evaluate the blood flow at different time points after operation. Then, the rats were sacrificed and the gastrocnemius and plasma were achieved to be further examined after angiography. Results Real-time PCR verified that the level of miR-221-3p was significantly decreased in the plasma, sclerotic samples or gastrocnemius compared with the controls no matter in the ASO patients or SD rat model. Conclusion MiR-221-3p could be a useful biomarker for ASO patients and may contribute to clinical decision making in treatments.