Background <p>Myocardial ischemia-reperfusion injury (IRI) remains a major clinical challenge, and sevoflurane (Sevo) preconditioning has shown cardioprotective effects. However, the underlying molecular mechanisms are incompletely understood. This study aimed to determine whether Sevo alleviates hypoxia/reoxygenation (H/R)-induced cardiomyocyte injury by regulating the lncRNA FAM201A.</p> Methods <p>To investigate the cardioprotective effects of Sevo, an optimized in vitro H/R model with Sevo preconditioning was established using H9c2 cardiomyocytes. FAM201A expression was modulated via overexpression plasmids, and miR-488-3p levels were altered using miRNA mimics in combination with Sevo exposure. Cell apoptosis, membrane integrity (cTnI, CK-MB), inflammatory responses (IL-6, TNF-α), and oxidative stress markers (SOD, MDA) were assessed. Bioinformatics prediction and dual-luciferase reporter assays validated interactions between FAM201A, miR-488-3p, and their target gene PTEN. Western blot analysis evaluated PI3K/AKT pathway activity.</p> Results <p>H/R significantly increased FAM201A expression in cardiomyocytes, which was dose-dependently suppressed by Sevo pretreatment. Overexpression of FAM201A abolished the protective effects of Sevo, as evidenced by exacerbated apoptosis rates, aggravated oxidative stress, and heightened inflammatory cytokine levels. Mechanistically, FAM201A directly interacted with and negatively regulated miR-488-3p. miR-488-3p mimics counteracted FAM201A-induced cellular damage and suppressed PTEN expression, a validated target of miR-488-3p. Furthermore, Sevo-mediated PTEN downregulation and subsequent AKT phosphorylation were reversed by FAM201A overexpression but rescued by miR-488-3p restoration.</p> Conclusions <p>Sevo preconditioning protects against H/R injury by downregulating FAM201A, which sponges miR-488-3p to reduce PTEN and activate PI3K/AKT signaling. Targeting this axis may provide potential therapeutic targets for IRI.</p>