Figure 6.

TMAO induces oxidative stress. Hippocampal slices were incubated in ACSF with either 0.03% DMSO (vehicle) or TMAO (50 μM) for 4 h and the lysates were analyzed for CCAAT-enhance-binding protein homologous protein (CHOP) protein expression and oxidative markers. (A) Representative western blot showing CHOP levels normalized to α-tubulin. (B) Hydrogen peroxide generation in the lysates was measured fluorometrically at Ex/Em 535/587 nm. (C) ROS generation was assayed using DCF dye and measured with a spectrophotometer. (D) Nitrite content was measured using Griess reagent and the azo product formed was measured spectrophotometrically at 545 nm. (E) Lipid peroxidation was estimated by measuring the malondialdehyde (MDA) content in the form of thiobarbituric acid reactive substances (TBARS method) spectrophotometrically. For (A) symbols/bars represent mean ± SEM; *indicates a significant difference between vehicle vs. TMAO treated slices, n = 3 mice per group, *p < 0.05. For (B–E) results are expressed as (%) change as compared to the control, Mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001.