Aim To observe the expression of miR-4463 in the plasma and tissues of patients with arteriosclerosis obliterans of lower limbs (ASO). Methods miRNA microarray method was used to screen the differentially expressed miRNA in the plasma of three ASO patients and three controls. Real time PCR technique was carried out to verify the expression level of miR-4463 in the plasma and tissues of 50 patients with ASO and 50 healthy controls, and the relationship of clinical stage with miR-4463 level was analyzed. Target gene prediction software was used to predict target genes of miR-4463, and then the function and signaling pathways of predicted target genes were analyzed by Gene Ontology and KEGG pathway. Results Compared with the controls, 51 miRNAs changed in the plasma of patients with ASO more than 1.5 times (P<0.05). The expression of miR-4463 in plasma and vascular intimal tissues of ASO patients was significantly decreased, and decreased gradually with Fontaine stage. Bioinformatics analysis revealed that the target genes of miR-4463 were related to cell polarity, migration, lipid metabolism and endocytosis. Conclusion miR-4463 is involved in the pathogenesis of ASO, and decreased miR-4463 level may indicate the occurrence of ASO.