Fig. 7
Effects of cPLA2 and iPLA2 inhibitors on levels of 4-HHE and 4-HNE in microglia cells treated with DHA and/or LPS. a Effects of cPLA2 inhibitors. Cells were pre-treated with U0126 (5 μM) or ATK (5 μM) for 1 h and followed by LPS (100 ng/mL) for 6 h. 4-HHE and 4-HNE levels were measured using the LC-MS/MS analysis as described in the “Methods” section. Each value represents the mean ± SEM of three biological replicates with duplicate analysis and is normalized to per 10 μg of protein. b Effects of iPLA2 inhibitor. Cells were pre-treated with BEL (5 μM) and /or DHA (50 μM) for 1 h prior to LPS (100 ng/mL) for 6 h. Results are representation of data from one cell passage; experiments repeated with different passages showed similar profiles. Analysis using one-way ANOVA followed with Bonferroni post-tests. In a, “a” represents significant increase (p < 0.001) in 4-HNE due to LPS stimulation as compared with control. In b, “a” represents significant increase (p < 0.01) in 4-HHE due to DHA and DHA + LPS compared with control and “b” represents significant decrease (p < 0.001) in 4-HHE due to BEL