dieldrin and Inflammation

Factors released from injured dopaminergic (DA) neurons may trigger microglial activation and set in motion a vicious cycle of neuronal injury and inflammation that fuels progressive DA neurodegeneration in Parkinson's disease. In this study, using proteomic and immunoblotting analysis, we detected elevated levels of cystatin C in conditioned media (CM) from 1-methyl-4-phenylpyridinium and dieldrin-injured rat DA neuronal cells. Immunodepletion of cystatin C significantly reduced the ability of DA neuronal CM to induce activation of rat microglial cells as determined by up-regulation of inducible nitric oxide synthase, production of free radicals and release of proinflammatory cytokines as well as activated microglia-mediated DA neurotoxicity. Treatment of the cystatin C-containing CM with enzymes that remove O- and sialic acid-, but not N-linked carbohydrate moieties markedly reduced the ability of the DA neuronal CM to activate microglia. Taken together, these results suggest that DA neuronal cystatin C plays a role in the neuronal injury-induced microglial activation and neurotoxicity. These findings from the rat DA neuron-microglia in vitro model may help guide continued investigation to define the precise role of cystatin C in the complex interplay among neurons and glia in the pathogenesis of Parkinson's disease.

Topics: Animals; Blotting, Western; Cell Survival; Cells, Cultured; Coloring Agents; Culture Media, Conditioned; Cystatin C; Cytokines; Dieldrin; Dopaminergic Neurons; Fluorescent Antibody Technique; Glycosylation; Inflammation; Macrophage Activation; Microglia; MPTP Poisoning; Mutagens; Neurons; Neurotoxicity Syndromes; Nitrites; Proteomics; Rats; Reactive Oxygen Species; Tetrazolium Salts; Thiazoles

We have recently found that dieldrin is a potent human neutrophil agonist in vitro and induces neutrophilic inflammation in vivo. Among the responses observed in vitro, dieldrin was found to induce superoxide (O2-) production by a yet unknown mechanism. In the present study, dieldrin- and phorbol 12-myristate 13-acetate (PMA)-induced O2- responses were compared. For this purpose, cells were preincubated with a panel of signal transduction inhibitors including genistein, H-7, HA-1077, pertussis toxin, staurosporine, calphostin C, SB203580, PD098059, and wortmannin. Dieldrin-induced O2- response was significantly reduced with treatment with genistein, H-7, HA-1077, staurosporine, and calphostin C, whereas PMA-induced response was significantly reduced by treatment with H-7, HA-1077, and staurosporine. This indicates that dieldrin mediates its effect via protein kinases C (PKCs) and tyrosine kinases. Involvement of tyrosine kinases in dieldrin-induced human neutrophils was further demonstrated by an increase in tyrosine phosphorylated protein level expression. Finally, we found that treatment with the mitochondrial stabilizer bongkrekic acid and with the inhibitor of vesicular transport brefeldin A did not reverse dieldrin-induced O2- response.

Topics: Cell Culture Techniques; Dieldrin; Humans; Inflammation; Insecticides; Neutrophils; Protein Kinase C; Protein-Tyrosine Kinases; Signal Transduction; Superoxides

Many chemicals of environmental concern are known to alter the immune system and are considered toxic molecules because they affect immune cell functions. Inflammation related to environmental chemical exposure, however, is poorly documented, except that from air pollutants. In this study, we found that the organochlorine insecticide dieldrin could not alter the ability of human neutrophils to phagocytose opsonized sheep red blood cells at nonnecrotic concentrations (0.1, 1, 10, and 50 microM). However, dieldrin was found to increase human neutrophil superoxide production, RNA synthesis, and proinflammatory cytokine interleukin-8 production. The normal apoptotic rate of neutrophils evaluated by both cytology and flow cytometry (CD-16 staining) was not altered by dieldrin treatments, and this was correlated with its inability to inhibit spreading of neutrophils onto glass. Using the murine air pouch model, we found that dieldrin induces a neutrophilic inflammation. Taken together, these results demonstrated that dieldrin is a proinflammatory contaminant. To our knowledge, this is the first report establishing that dieldrin is a contaminant exhibiting proinflammatory properties. In addition, it is the first time that the murine air pouch model has been successfully used to confirm that a chemical of environmental concern can induce an inflammatory response in vivo.

Topics: Animals; Apoptosis; Cell Adhesion; Cells, Cultured; Dieldrin; Humans; Inflammation; Interleukin-8; Mice; Mice, Inbred C57BL; Neutrophil Activation; Neutrophil Infiltration; Neutrophils; Phagocytosis; RNA; Superoxides