Figure 1.
Effect of TFA on endothelial NF-κB signaling.
Human endothelial cells were treated with increasing concentrations of palmitate, linoleic acid, transvaccenic (trans-C18:1 (11 trans)), linoelaidic (trans-C18:2 (9 trans, 12 trans)), and elaidic (trans-C18:1 (9 trans)) for 3 h. All fatty acids were initially complexed with BSA and in the control condition cells were treated with BSA alone for 3 h. A. Phospho-IκBα levels were measured using an ELISA assay. Fold-increase over the control (BSA alone) condition was calculated. (n = 3, *p<0.05). B. IL-6 levels as measured by ELISA. (n = 3, * p<0.05). C. Expression levels of TNFα, ICAM, and iNOS in response to 100 µM of fat. (n = 3, * p<0.05).
Figure 2.
Effect of TFA on endothelial insulin signaling.
Human endothelial cells were treated with either 100 µM of palmitate, linoleic acid, transvaccenic (trans-C18:1 (11 trans)), linoelaidic (trans-C18:2 (9 trans, 12 trans)), and elaidic (trans-C18:1 (9 trans)) for 3 h and then stimulated with insulin 100 nM or vehicle for 15 min. A. Cell lysates were made and were analyzed for insulin-mediated phosphorylation of serine 473 Akt and total Akt by ELISA. Fold increase over vehicle condition was calculated. (n = 3, *p<0.05) B. Cell lysates were analyzed for insulin-mediated phosphorylation of serine 1177 eNOS and total eNOS by Western blot. Representative phospho-eNOS Western blots are shown. Fold increase over vehicle condition was calculated (n = 3, *p<0.05). C. Insulin-mediated nitric oxide production was measured by ESR using the spin trap Fe(DETC)2 (n = 3, *p<0.05).
Figure 3.
Effect of TFA on endothelial superoxide production.
Human endothelial cells were treated with either 100 µM of palmitate, linoleic acid, transvaccenic (trans-C18:1 (11 trans)), linoelaidic (trans-C18:2 (9 trans, 12 trans)), and elaidic (trans-C18:1 (9 trans)) for 3 h. A. Superoxide levels were measured by ESR and the spin trap CMH. Fold increase over the control condition (BSA alone) was calculated. (n = 3, *p<0.05, compared to BSA control). B. Cells were pretreated with either of two inhibitors of superoxide (SOD, 100 Units/ml or DPI 25 µM) prior to stimulation with Elaidic or Linoelaidic acid. IL-6 levels as measured by ELISA.