Folia Biologica
Journal of Cellular and Molecular Biology, Charles University 

Nerol (NRL), a monoterpene in essential oils of various plants, exhibits anticancer activity, albeit with contrasting effects on normal cells. Because anaemia is induced by anticancer drugs, this study was initiated to profile the cytotoxic mechanisms of NRL in human erythrocytes. Cells were treated with NRL (0.01–0.1 %) for 24 h at 37 °C in Ringer buffers. Lytic cell death was measured photometrically and eryptosis was examined by flow cytometry using forward scatter, annexin V-affinity assay, H₂DCFDA, and Fluo4/AM to evaluate cell volume, phosphatidylserine (PS) externalization, oxidative stress, and Ca²⁺, respectively. Small molecule inhibitors were used to probe the molecular mechanisms governing NRL-induced cytotoxicity. NRL led to eryptotic volume loss associa­ted with increased PS externalization and lysis through oxidative stress and Ca²⁺ nucleation. Ca²⁺ deprivation and K⁺ gradient dissipation, along with glucose, guanosine and sucrose, significantly blunted NRL-induced eryptosis and lytic death. Notably, both forms of cell death were inhibited by staurosporine, necrostatin 2 and myriocin, whereas urea, Z-VAD-FMK, SB205830 and D4476 only attenuated eryptosis. Furthermore, while lytic death was inhibited by polyethylene glycol, it was potentiated by Ca²⁺ deprivation and heparin, whereas eryptosis was aggravated by uric acid. In conclusion, NRL triggers cation channel- and redox-mediated eryptosis and lytic death through energy deprivation and activation of protein kinase C, receptor-interacting protein 1, serine palmitoyltransferase, caspase, p38 MAPK and casein kinase 1α. Altogether, these findings underscore the differential mechanisms by which NRL modulates divergent erythrocyte injury pathways and collectively advance the current knowledge of the extent of its cellular effects.

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