Fig. 4.

Ethanol increased oxidative stress and phagocytic dysfunction in mouse alveolar macrophages (mAMs). In cultured MH-S cells that were either untreated (Con) or ethanol-treated (EtOH, 0.08%) for 3 days ± GSH for either the last 24 h (GSH, 1 day) or for the duration of ethanol exposure (GSH, 3 days) (n = 3, in duplicate), intracellular reactive oxygen species (ROS) production was measured as 2′,7′-dichlorofluorescein (DCF) fluorescence (A), and extracellular H₂O₂ was measured by Amplex Red assay (B). Phagocytic ability of these MH-S cells was assessed by phagocytosis assay (n = 3, 10 fields/experimental condition) (C). In mAMs collected from control (Con) and ethanol-fed (EtOH) mice ± GSH for the last 7 days of ethanol feeding (n = 5, in duplicate), DCF fluorescence (D) and H₂O₂ generation (E) were measured. Phagocytic ability of these mAMs was assessed by phagocytosis assay (n = 5, 10 fields/experimental condition) (F). Phagocytic index was calculated from the percentage of phagocytic cells multiplied by the relative fluorescence units of Staphylococcus aureus per cell. All values are expressed as means ± SE, relative to control. *P < 0.05 vs. Con; ^ΦP < 0.05 vs. EtOH.