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- Cardiotoxic effects of methylone in rat H9c2 cardiomyocytes: role of oxidative stress and antioxidant interventionPublication . Moreira, Maria Monteiro; Carvalho, Márcia; Araújo, Ana MargaridaSynthetic cathinones (SCs) have emerged in recreational contexts as a novel class of psychoactive substances, producing amphetamine-like effects but also severe cardiovascular complications, including myocardial infarction and sudden cardiac death. Despite these risks, the mechanisms underlying SC-induced cardiotoxicity remain poorly understood. This study aimed to investigate the cardiotoxic potential of methylone, one of the most widely abused first-generation SCs. H9c2 rat cardiomyoblasts were exposed to methylone (0.01 - 4.0 mM) for 24 and 48 hours. Cell viability was determined by 3- (4,5-Dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) reduction assay, and intracellular levels of reactive oxygen/nitrogen species (ROS/RNS) were quantified using the 2′,7′-dichlorofluorescein diacetate probe. The protective effects of the antioxidants ascorbic acid, N-acetyl-L-cysteine, and Trolox were also evaluated. Methylone induced concentration-dependent cytotoxicity, with an IC50 of 0.98 mM. ROS/RNS production increased markedly in both a concentration- and time-dependent manner, reaching a 4.8- fold increase after 24 h at IC50. Among the antioxidants tested, only ascorbic acid significantly improved cell viability. These results demonstrate that methylone exerts cardiotoxic effects in vitro, with oxidative stress playing a central role in its toxicity. The partial protection conferred by ascorbic acid highlights its potential as a therapeutic candidate against SC-induced cardiac injury.