Background In this study, we aimed to investigate the role of miR-26a and miR-26b in the management of AF. Methods Real-time PCR was carried out to determine plasma microRNA expression in AF patients pre- and post-radiofrequency ablation. The correlation between the expression of SELP and miR-26a/miR-26b was also studied using luciferase assays to establish a miR-26a/miR-26b/SELP signaling pathway. Results The relative expression of SELP reached its peak in pre-ablation AF ( +) patients, while ablation treatment reduced the expression of SELP in AF ( +) patients. Similarly, AF pigs showed dysregulation of miR-26a/b and SELP, thus verifying the involvement of miR-26a/b and SELP in AF. Meanwhile, the regulatory association between SELP and miR-26a/b was also investigated, and the results showed that the presence of pre-miR-26a/b increased the levels of miR-26a/b and inhibited the mRNA/protein expression of SELP. Finally, using bioinformatic tools and luciferase assays, SELP mRNA was confirmed as the target of miR-26a/b, which affected the effect of AF ablation treatment. Conclusions RFA helped to restore circulating levels of miR-26, which were reduced in atrial fibrillation. Meanwhile, miR-26 is a potential cause for the elevated plasma levels of pro-thrombogenic SELP in that disease.