Fig. 2

Acute melatonin attenuated the acute ethanol-induced increase in ROS and oxidative stress in the rat pups and in HT22 and BV2 cells. a A representative histogram of the ROS level in the brain homogenates of the rat pups. b A representative histogram of the MDA level in the brain homogenates of the rat pups. n = 10 pups/group, and the number of experiments = 3. c Representative image of immunofluorescence staining of 8-OxoG in the cortices and CA1 regions of the hippocampi in the rat pups. n = 5 pups/group, and the number of experiments = 3. Magnification × 40. Scale bar = 50 μm. d Representative images of the DCF immunofluorescence intensity in the HT22 cells. The number of experiments = 3. Magnification × 40. Scale bar = 50 μm. e, f A representative histogram of the relative absorbance of ROS-positive HT22 and BV2 cells respectively that were subjected to Nrf2 siRNA and treated with ethanol (100 mM) and melatonin (100 μM) for 12 h. The number of experiments = 3. The data are expressed as the mean ± SEM. The data are presented relative to control values. Significance = P < 0.05. σ Significantly different from the control saline-treated rat pups; Φ significantly different from the ethanol-treated rat pups. Similarly for in vitro studies, σ significantly different from the non-treated HT22 and BV2 cells, Φ significantly different from the ethanol-exposed HT22 and BV2 cells, and ω significantly different from the ethanol + melatonin-exposed HT22 and BV2 cells