Asthenozoospermia is detected in 40% of infertile men, and characterised by low sperm motility. MicroRNAs (miRNAs) play essential roles in spermatogenesis, but little is known regarding the function of seminal plasma miRNAs in asthenozoospermia. In this study, we collected seminal plasma samples from patients with asthenospermia and healthy men and employed high-throughput sequence technology to identify differentially expressed miRNAs. Thirteen altered miRNAs were confirmed by qRT-PCR. Six of these miRNAs were upregulated, and seven were downregulated. Five of the miRNAs (hsa-miR-34c-5p, hsa-miR-34b-5p, hsa-miR-146b-5p, hsa-miR-449a and has-miR-765) had been characterised previously, and eight of the others (miR-5000-3p, miR-4289, miR-6514-3p, miR-6882-5p and miR-6739-5p, miR-135a-5p, miR-509-3p and miR-196b-5p) were identified in asthenospermia for the first time in this study. These miRNAs were significantly associated with PI3K-Akt signaling pathway, MAPK signaling pathway, HIF-1 signaling pathway and FoxO signaling pathway. The identified dysregulated miRNA may be the key to the development of new and enhanced diagnosis and prognosis technologies for asthenospermia, and may also provide new therapeutic possibilities in the field of personalised medicine.