Fig. 2

Effects of DHA, 4-HHE, and 4-HNE on LPS-induced NO production in BV-2 microglial cells. BV-2 microglial cells (10⁶) were subculture in 96-well plates to 80% confluent. At the time of experiment, cells were serum-starved for 3 h and pre-treated with DHA (12.5–100 μM), 4-HHE (1.25–10 μM), and 4-HNE (1.25–10 μM) for 1 h, and followed by stimulation with LPS (100 ng/mL) for 16 h. Data in a–c represent inhibition of LPS-induced NO production by DHA, 4-HHE, and 4-HNE using the Griess reagent. Results were obtained from triplicate assay from each cell passage. Concentrations of NO in μM ± SE (n = 5) were as follows: DHA 9.75 ± 1.8, 4-HHE 9.30 ± 0.10, and 4-HNE 10.43 ± 0. 52. Data in d–f represent cell viability after 16-h incubation using the WST-1 assay. Results are expressed as the mean ± SEM (n = 3–5) and analyzed by one-way ANOVA followed by Bonferroni post-tests; “a” represents significant differences (p < 0.05) comparing test compounds with control (Ctrl) with LPS treatment alone. IC₅₀ values for each test compound were determined using the formula for regression analysis in Microsoft Excel 2016