Melatonin Pre-Treatment Protects Erythrocytes Against Subsequent Oxidative Damage

Research on the effects of melatonin on erythrocyte deformability has yielded mixed results. While some studies reported improvements, others found no effect, and a few even noted a deterioration in deformability. Moreover, the impact of melatonin may vary between healthy erythrocytes and those subjected to oxidative stress. This study investigated the dose-dependent effects of melatonin on erythrocytes under baseline conditions and oxidative stress, using both pre- and post-stress incubation protocols. Oxidative damage was induced with tert-butyl hydroperoxide (TBHP), and its extent was assessed via dichlorofluorescein fluorescence. Erythrocyte deformability was measured using ektacytometry, and osmotic resistance was assessed through hemolytic assays. The results showed that incubation with TBHP led to a dose-dependent decline in both erythrocyte deformability and osmotic resistance. While melatonin treatment had no observable effect on intact erythrocytes, it enhanced deformability in oxidatively damaged erythrocytes when administered before oxidative stress was induced. However, the beneficial effect was not evident when melatonin was applied after oxidative damage. Additionally, melatonin incubation had no impact on the ability of erythrocytes to resist the hypotonic environment. In conclusion, this study supports the notion that the antioxidant properties of melatonin can improve erythrocyte functional status, as reflected by enhanced deformability, but not osmotic resistance. Notably, this effect was observed only in erythrocytes that were exposed to oxidative damage after melatonin incubation, not in intact cells.