5-Lipoxygenase Inhibition Protects Retinal Pigment Epithelium from Sodium Iodate-Induced Ferroptosis and Prevents Retinal Degeneration

Figure 2

Inhibition of lipoxygenase-5 (5-LOX) protected ARPE-19 cells from NaIO₃-induced cell death by ferroptosis. (a) Arachidonic acid (ARA, black arrow) and 5-hydroxyeicosatetraenoic acid (5-HETE, white arrow) were detected with HPLC-MS/MS analysis. CCK-8 was used to analyze cell viability following treatments (b–f). Cell viability of ARPE-19 following treatment with (b) nonlethal concentration of NaIO₃ (10 mM, 20 h) and different concentrations of arachidonic acid, (c) toxic NaIO₃ (35 mM, 20 h) and different concentrations of 5-LOX inhibitor Zileuton ( per treatment condition), and (d) other types of LOX inhibitors (75 μM Zileuton, 15 μM Mk-886, 3 μM ML355, 500 nM BLX-3887, per treatment condition). (e) The effect of NaIO₃ (35 mM, 20 h) with ALOX5 knockdown-ARPE-19 cell. (f) Effect of combined treatment with suboptimal concentrations of Zileuton (40 μM) and DFO (25 μM) on NaIO₃ (35 mM, 20 h)-induced ferroptosis. Abbreviations: HPLC-MS/MS: high-performance liquid chromatography with tandem mass spectrometry; PBS: phosphate-buffered saline; sc: scramble; KD: knockdown.

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