epidermal-growth-factor and linsidomine

Epidermal growth factor (EGF) has been considered to be a candidate for neurotrophic factors on the basis of the results of several in vitro studies. However, the in vivo effect of EGF on ischemic neurons as well as its mechanism of action have not been fully understood. In the present in vivo study using a gerbil ischemia-model, we examined the effects of EGF on ischemia-induced learning disability and hippocampal CA1 neuron damage. Cerebroventricular infusion of EGF (24 or 120 ng/d) for 7 days to gerbils starting 2 hours before or immediately after transient forebrain ischemia caused a significant prolongation of response latency time in a passive avoidance task in comparison with the response latency of vehicle-treated ischemic animals. Subsequent histologic examinations showed that EGF effectively prevented delayed neuronal death of CA1 neurons in the stratum pyramidale and preserved synapses intact within the strata moleculare, radiatum, and oriens of the hippocampal CA1 region. In situ detection of DNA fragmentation (TUNEL staining) revealed that ischemic animals infused with EGF contained fewer TUNEL-positive neurons in the hippocampal CA1 field than those infused with vehicle alone at the seventh day after ischemia. In primary hippocampal cultures, EGF (0.048 to 6.0 ng/mL) extended the survival of cultured neurons, facilitated neurite outgrowth, and prevented neuronal damage caused by the hydroxyl radical-producing agent FeSO4 and by the peroxynitrite-producing agent 3-morpholinosydnonimine in a dose-dependent manner. Moreover, EGF significantly attenuated FeSO4-induced lipid peroxidation of cultured neurons. These findings suggest that EGF has a neuroprotective effect on ischemic hippocampal neurons in vivo possibly through inhibition of free radical neurotoxicity and lipid peroxidation.

Topics: Animals; Apoptosis; Avoidance Learning; Brain Ischemia; Cells, Cultured; DNA Fragmentation; Epidermal Growth Factor; Ferrous Compounds; Free Radicals; Gerbillinae; Hippocampus; Lipid Peroxidation; Male; Molsidomine; Neurites; Neurons; Neuroprotective Agents; Neurotoxins; Oxidative Stress; Prosencephalon; Reaction Time; Synapses

Bovine retinal pigmented epithelial (RPE) cells express, after activation with interferon gamma (IFN-gamma) and lipopolysaccharide (LPS), an inducible nitric oxide synthase (NOS). Experiments were done to investigate the effects of the transforming growth factor beta 1, epidermal growth factor, and fibroblast growth factors (FGFs), which are abundant in the retina, on NOS activity. Transforming growth factor beta 1 slightly increases the production of nitrite, an oxidation product of NO, induced by LPS plus IFN-gamma, whereas acidic and basic FGFs markedly inhibit the nitrite release due to LPS/IFN-gamma in a concentration-dependent manner, and epidermal growth factor did not modify LPS/IFN-gamma-induced NOS activity. The growth factors alone did not stimulate nitrite release. We have attempted to elucidate the mechanism of FGF inhibition. Results with heparin, suramin, and tyrphostin suggest involvement of the high-affinity receptor for FGF in its inhibition of LPS/IFN-gamma-stimulated NOS activity. Continued stimulation of RPE cells with LPS/IFN-gamma was essential for the induction of NO synthesis, and maximal inhibition was obtained when FGF was present during stimulation with LPS/IFN-gamma, suggesting that FGF inhibits NOS induction. Furthermore, an antiproliferative action of NO was demonstrated by an inverse correlation between the amounts of nitrite or citrulline produced in response to different stimuli (LPS/IFN-gamma or LPS/IFN-gamma with growth factors) and the level of cellular proliferation. Similar inhibition of growth was obtained when RPE cells were incubated with an NO donor, sydnonimide. Because NO acts as a cytotoxic compound in the retina, FGF, by inhibiting the induction of NOS in RPE cells, may have beneficial effects in protecting the retina from cytokine and endotoxin-mediated tissue damage.

Topics: Amino Acid Oxidoreductases; Animals; Catechols; Cattle; Cells, Cultured; Enzyme Induction; Epidermal Growth Factor; Fibroblast Growth Factor 1; Fibroblast Growth Factor 2; Heparin; Humans; Interferon-gamma; Kinetics; Lipopolysaccharides; Molsidomine; Nitric Oxide Synthase; Nitriles; Nitrites; Pigment Epithelium of Eye; Recombinant Proteins; Suramin; Transforming Growth Factor beta; Tyrphostins; Vasodilator Agents