Effect of 2,4-bis( p -hydroxyphenyl)-2-butenal on LPS-induced ROS and NO release, and expression of cytokines in astrocytes and microglial BV-2 cells. Cells were treated with 1 μg/ml of LPS alone, or with LPS plus different concentrations (1, 2, 5 μM) of 2,4-bis(p-hydroxyphenyl)-2-butenal, at 37°C for 24 h. Intracellular ROS levels were determined by measuring DCF fluorescence (A). NO level was determined by Griess reaction, as described in Methods, in supernatants from astrocytes (B) and microglial BV-2 cells (C). mRNA levels for TNF-α and IL-1β were determined by real-time PCR, as described in Methods, in astrocytes (D) and microglial BV-2 cells (E). Values represent means ± SD for three independent experiments performed in triplicate. * indicates significantly different from LPS treated group (P < 0.05).