Fig. 5

Acute melatonin reduced the level of activated gliosis in the acute ethanol-treated rat pups and activated microglia in the BV2 cells that were exposed to ethanol. a Western blot analysis of GFAP and Iba-1 in the rat pups. The bands were quantified using Sigma Gel software, and the differences are presented in a histogram. β-actin was used as a loading control. n = 10 pups/group, and the number of experiments = 3. b, c Representative images showing the immunofluorescence analysis of GFAP and Iba-1 in the cortices and the CA1 regions of the hippocampi in the rat pups, respectively. n = 5 pups/group, and the number of experiments = 3. Magnification × 40. Scale bar = 50 μm. d Western blots and the densitometric analysis of Iba-1 expression in the BV2 cells that were subjected to Nrf2 siRNA and treated with ethanol (100 mM) and melatonin (100 μM) for 12 h. β-Actin was used as a loading control. 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 BV2 cells; Φ significantly different from the ethanol-exposed BV2 cells, and ω significantly different from the ethanol + melatonin-exposed BV2 cells