Figure 3.

Ethanol mediated the expression of AD markers and proinflammatory cytokines in a time-dependent manner in HPAs. qPCR (A–F) analysis showing the time-dependent (3–96 h) mRNA expression of APP (A), BACE1 (B), BACE1-AS (C), and the proinflammatory cytokines TNFα (D), IL6 (E), and IL1β (F) in HPAs exposed to ethanol at a 12.5 mM dose for the indicated times. GAPDH was used as an internal control. Western blots (G–I) showing the time-dependent upregulation of APP (G), Aβ mOC64 (H), and BACE1 (I) proteins in HPAs following the exposure of cells to ethanol (12.5 mM) for the indicated times. For the APP and Aβ mOC64, a single membrane was divided into two halves: the upper half was probed with the APP antibody, and the lower half was probed with the Aβ mOC64 antibody; β-actin was similar for both of these blots. β-Actin was used as an internal control for normalization. Data are presented as mean ± SEM; n = six biological replicates per group. One-way ANOVA followed by Tukey’s post hoc test was used to determine the statistical significance: * p < 0.05 versus control. Abbreviations: APP, amyloid precursor protein; Aβ, amyloid beta; BACE1, β-site-cleaving enzyme; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; BACE1-AS, BACE1-antisense transcript; TNFα, tumor necrosis factor α; IL6, interleukin-6; IL1β, interleukin-1β; HPA, human primary astrocyte; qPCR, quantitative polymerase chain reaction.