A FABP4-PPARγ signaling axis regulates human monocyte responses to electrophilic fatty acid nitroalkenes.

Nitro-fatty acids (NO2-FA) are electrophilic lipid mediators derived from unsaturated fatty acid nitration. These species are produced endogenously by metabolic and inflammatory reactions and mediate anti-oxidative and anti-inflammatory responses. NO2-FA have been postulated as partial agonists of the Peroxisome Proliferator-Activated Receptor gamma (PPARγ), which is predominantly expressed in adipocytes and myeloid cells. Herein, we explored molecular and cellular events associated with PPARγ activation by NO2-FA in monocytes and macrophages. NO2-FA induced the expression of two PPARγ reporter genes, Fatty Acid Binding Protein 4 (FABP4) and the scavenger receptor CD36, at early stages of monocyte differentiation into macrophages. These responses were inhibited by the specific PPARγ inhibitor GW9662. Attenuated NO2-FA effects on PPARγ signaling were observed once cells were differentiated into macrophages, with a significant but lower FABP4 upregulation, and no induction of CD36. Using in vitro and in silico approaches, we demonstrated that NO2-FA bind to FABP4. Furthermore, the inhibition of monocyte FA binding by FABP4 diminished NO2-FA-induced upregulation of reporter genes that are transcriptionally regulated by PPARγ, Keap1/Nrf2 and HSF1, indicating that FABP4 inhibition mitigates NO2-FA signaling actions. Overall, our results affirm that NO2-FA activate PPARγ in monocytes and upregulate FABP4 expression, thus promoting a positive amplification loop for the downstream signaling actions of this mediator.

Density functional study of isomerization of fluoro- and chloroformaldehyde radical cations.

Ab initio and density functional (DFT) calculations were performed on radical cations with the formula HXCO(+˙) (X = H, F, and CI) and their isomers XCOH(+˙) . Hartree-Fock, Møller-Plesset at second order (MP2), and quadratic configuration interaction including singles and doubles (QCISD) methods were employed for geometry optimizations at the ab initio level. Becke's 1988 and three-parameter exchange potentials, together with Vosko-Wilk-Nusair (VWN) and Lee-Yang-Paar (LYP) correlation potentials (BVWN, BLYP, and B3LYP) were used for the DFT calculations. HF and MPn isomerization energies are severely in error, mostly due to a bad description of the XHCO(+˙) cation radicals at the Hartree-Fock level, leading to oscillatory behavior of the perturbation series. QCISD methods, at least, are needed to obtain accurate results at the conventional ab initio level, even using large extended basis sets. B3LYP results are most similar to QCISD results for the isomerization energies of the three cation radicals. Parent neutral species are also described to a high degree of accuracy. B3LYP methods are faster than QCISD (and as exact as them) for all the cases studied here. MP2 methods, although giving incorrect results, are faster than B3LYP up to about 80 basis functions. For larger problems, B3LYP methods are faster. The best theoretical results obtained indicate that fluoro- and chloroformaldehyde cation radicals are less stable than their hydroxyfluoro- and hydroxychloromethylene isomers, the reverse situation than for the formaldehyde cation radical. The best values found in this article for the isomerization energy of XHCO(+˙) are -26 ± 2 kJ/mol (X = H), +37 ± 2 kJ/mol (X = F), and +28 ± 2 kJ/mol (X = CI). Ionization energies of 10.9, 12.3, and 11.4 eV are predicted for the XHCO species (X = H, F, CI). © 1996 by John Wiley & Sons, Inc.