Objective <p>Myocardial ischemia-reperfusion injury (MIRI) is a major contributor to global disease mortality. This study aims to investigate the protective mechanism of the RMRP/miR-206 axis in MIRI following sevoflurane (Sev) preconditioning.</p> Methods <p>In this study, a hypoxia/reoxygenation (H/R) model was constructed by H9c2 cells. The optimal concentration of Sev was selected by assessing cell viability with a CCK-8 assay. After Sev preconditioning, flow cytometry was employed to detect cell apoptosis. The content of LDH was measured with a lactate dehydrogenase (LDH) assay kit. The SOD activity was examined by an SOD assay kit, and the levels of IL-6 and IL-10 were determined by enzyme-linked immunosorbent assay (ELISA). RT-qPCR was utilized to detect the RMRP and miR-206 expression. The luciferase reporter assay and RIP assay were employed to demonstrate the interaction between RMRP and miR-206.</p> Results <p>In this study, 2.0% was chosen as the optimal Sev concentration for subsequent experiments. After Sev preconditioning, cell viability increased, apoptosis decreased, LDH levels were reduced, SOD activity was enhanced, IL-6 was downregulated, and IL-10 was upregulated. Meanwhile, RMRP levels decreased significantly after Sev preconditioning. Overexpression of RMRP inhibited the protective effect of Sev preconditioning on H/R-induced cells. Further analysis revealed that RMRP targeted miR-206. While miR-206 increased after Sev preconditioning, RMRP transfection suppressed miR-206 expression. Overexpression of miR-206 rescued H/R-induced cell damage caused by RMRP upregulation.</p> Conclusion <p>Sev preconditioning protects cells from MIRI by regulating the RMRP/miR-206 axis.</p> Clinical trial number <p>Not applicable.</p>