kt-5720 and Glioma

The CNS neurotoxic effects of lead (Pb) are well documented; however, the molecular toxicity targets have not been clearly delineated. Astroglial cells, which are the most abundant cells in the brain and provide critical support to the neurons, are known to accumulate Pb. Although NO generated by inducible NO synthase (iNOS) in glial cells has been associated with many neurotoxic events, it can also serve to protect by modulating blood flow, increase antimicrobial and tumoricidal activities, and promote immune responses following injury or insult. The present investigations were designed to test the hypothesis that Pb exposure may perturb cytokine signal transduction pathways leading to NO production by astroglial cells. Pretreatment with Pb acetate (500 nM-10 microM) attenuated the generation of NO in a concentration-dependent manner up to 90%, and suppressed iNOS protein expression, as well as interfered with the homeostatic functions of calcium in the cytokine-induced NO signal transduction pathway. In addition, pretreatment with staurosporine, a serine-threonine kinase inhibitor, or KT5720, a specific protein kinase A inhibitor (PKA), inhibited cytokine-induced NO production in a concentration-dependent manner with IC(50) values of 26.3 and 346.7 nM, respectively. Therefore, Pb may impede events within the PKA signal transduction pathway; although, based on results from a gel shift assay, Pb does not directly affect PKA enzyme activity. Taken together, these results suggest the possibility that the suppressive effect of Pb acetate on cytokine-induced NO production in glial cells may be implicated in the neurophysiologic changes noted following occupational or environmental exposure to Pb.

Topics: Animals; Carbazoles; Cytokines; Dose-Response Relationship, Drug; Drug Combinations; Enzyme Inhibitors; Glioma; Indoles; Neuroglia; Nitric Oxide; Organometallic Compounds; Protein Kinase Inhibitors; Pyrroles; Rats; Signal Transduction; Staurosporine; Tumor Cells, Cultured