Physiological Concentration of Exogenous Lactate Reduces Antimycin A Triggered Oxidative Stress in Intestinal Epithelial Cell Line IPEC-1 and IPEC-J2 In Vitro (original) (raw)
Fig 1
Normalised Anti A triggered ROS generation in IPEC cells after lactate and glucose treatment.
(A) IPEC-1 cells were cultured on glass (7 d), incubated for 1 h in buffer (HBSS), placed on microscopic stage and challenged with Anti A in the presence of DHE. Mean fluorescence of nuclei (regions of interest, white circles) was monitored over time. (B) Quantification of nucleic fluorescence of ethidium cation (ethidium cation is the oxidation product of DHE) binding to nuclei. Anti A (10 / 100 μM) or control buffer was applied in presence of DHE 1.5 min after starting the measurement. (C) Anti A (100 μM) triggered superoxide generation rate (DHE oxidation) of IPEC-1 and IPEC-J2 cells after 1 h treatment (39°C). Cells were treated with lactate (lac, 25 mM); lac + inhibitor of monocarboxylate transporter (MCT inhibitor, ARC155858, 10 μM); increased glucose (gluc, 4.5 g/L) or glycolysis inhibitor 2-Deoxyglucose (2DG, 10 mM). Glucose concentration was 1 g/L if not otherwise indicated. Slope of fluorescence increase of individual cell nuclei was calculated and normalised to the control. Statistical analysis was done by Kruskal-Wallis and Dunn's post-hoc test. Different superscripts indicate statistically significant (p<0.05) differences.