dihydropyridines and acetovanillone

Platelet-activating factor (PAF) is produced by macrophages during inflammation and infections. We evaluated whether PAF is able to modulate the infection of human macrophages by Leishmania braziliensis, the main Leishmania sp. in Brazil. Monocyte-derived macrophages were incubated with promastigote forms in absence or presence of exogenous PAF. We observed that the treatment of macrophages with low concentrations of PAF prior to infection increased the phagocytosis of L. braziliensis. More importantly, exogenous PAF reduced the parasitism when it was added before, during or after infection. In addition, treatment with a PAF antagonist (PCA 4248) resulted in a significant increase of macrophage infection in a concentration-dependent manner, suggesting that endogenous PAF is important to control L. braziliensis infection. Mechanistically, while exogenous PAF increased production of reactive oxygen species (ROS) treatment with PCA 4248 reduced oxidative burst during L. braziliensis infection. The microbicidal effects of exogenous PAF were abolished when macrophages were treated with apocynin, an NADPH oxidase inhibitor. The data show that PAF promotes the production of ROS induced by L. braziliensis, suggesting that this lipid mediator may be relevant to control L. braziliensis infection in human macrophages.

Topics: Acetophenones; Dihydropyridines; Enzyme Inhibitors; Gene Expression; Humans; Leishmania braziliensis; Macrophages; NADPH Oxidases; Phagocytosis; Platelet Activating Factor; Primary Cell Culture; Reactive Oxygen Species; Respiratory Burst

Insulin resistance combined with hyperinsulinemia is involved in the generation of oxidative stress. There is known to be a relationship between increased production of reactive oxygen species and the diverse pathogenic mechanisms involved in diabetic vascular complications including nephropathy. The present study found that high doses of insulin affect mesangial cell proliferation through the generation of intracellular reactive oxygen species and the activation of cell signaling pathways. We also examined whether azelnidipine, a dihydropyridine-based calcium antagonist with established antioxidant activity, has the potential to inhibit mesangial cell proliferation. Cell proliferation was increased in a dose-dependent manner by high doses of insulin (0.1-10 microM), but was inhibited by 0.1 microM azelnidipine. Phosphorylation of extracellular signal-regulated kinase (ERK)-1/2 was found to be increased by insulin in a dose-dependent manner (0.1-10 microM). This increased phosphorylation of ERK-1/2 was inhibited by treatment with 0.1 microM azelnidipine. Intracellular oxidative stress was also increased by insulin stimulation in a dose-dependent manner (0.01-10 microM), and 0.1 microM azelnidipine was found to block intracellular reactive oxygen species production more effectively than 0.1 microM nifedipine. The NAD(P)H oxidase inhibitor, apocynin (0.01-0.1 microM), prevented insulin-induced mesangial cell proliferation. Taken together, these results suggest that azelnidipine inhibits insulin-induced mesangial cell proliferation by inhibiting the production of reactive oxygen species. Given these pharmacological characteristics, azelnidipine may have the potential to protect against the onset of diabetic nephropathy and slow its progression.

Topics: Acetophenones; Animals; Antioxidants; Azetidinecarboxylic Acid; Blotting, Western; Calcium Channel Blockers; Cell Proliferation; Dihydropyridines; DNA; Extracellular Signal-Regulated MAP Kinases; Hypoglycemic Agents; Insulin; Mesangial Cells; Mitogen-Activated Protein Kinase 1; Oxidative Stress; Phosphorylation; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species