FIGURE 8.

PIO reverses alcohol-induced derangements that cause phagocytic dysfunction in hAM. hAM were collected from control subjects (Con; n = 17) and from subjects with an AUD (Alc; n = 17). hAM were then treated ± PIO (10 μM) ex vivo for 1 d. (A) mRNA levels of Nox1, Nox2, and Nox4 were measured in hAM by quantitative RT-PCR (qRT-PCR; in duplicate), normalized to 9s mRNA. Protein expression of Nox1, Nox2, and Nox4 were determined in hAM (10 fields per condition) using fluorescence microscopy. Fluorescence was normalized to DAPI nuclear stain and expressed as mean RFU per cell. (B) hAM ROS production was measured by DCFH-DA fluorescence assay and expressed as mean RFU per cell. (C) H₂O₂ generation was measured by Amplex Red assay (in duplicate) and normalized to protein concentration in the same sample. (D) hAM phagocytic ability was assessed by phagocytosis assay (10 fields per condition). Phagocytic index was calculated from the percentage of cells positive for bacterial uptake multiplied by the RFU of S. aureus per cell. All values are expressed as mean ± SEM, relative to control. *p < 0.05 versus Con, ^Φp < 0.05 versus Alc.