Modulation of ROS production in human leukocytes by ganglioside micelles

Recent studies have reported that exogenous gangliosides, the sialic acid-containing glycosphingolipids, are able to modulate many cellular functions. We examined the effect of micelles of mono- and trisialoganglioside GM1 and GT1b on the production of reactive oxygen species by stimulated human polymorphonuclear neutrophils using different spectroscopic methods. The results indicated that exogenous gangliosides did not influence extracellular superoxide anion (O2.-) generation by polymorphonuclear neutrophils activated by receptor-dependent formyl-methionyl-leucyl-phenylalanine. However, when neutrophils were stimulated by receptor-bypassing phorbol 12-myristate 13-acetate (PMA), gangliosides above their critical micellar concentrations prolonged the lag time preceding the production in a concentration-dependent way, without affecting total extracellular O2.- generation detected by superoxide dismutase-inhibitable cytochrome c reduction. The effect of ganglioside GT1b (100 µM) on the increase in lag time was shown to be significant by means of both superoxide dismutase-inhibitable cytochrome c reduction assay and electron paramagnetic resonance spectroscopy (P < 0.0001 and P < 0.005, respectively). The observed phenomena can be attributed to the ability of ganglioside micelles attached to the cell surface to slow down PMA uptake, thus increasing the diffusion barrier and consequently delaying membrane events responsible for PMA-stimulated O2.- production.

Superoxide release and cellular gluthatione peroxidase activity in leukocytes from children with persistent asthma

Asthma is an inflammatory condition characterized by the involvement of several mediators, including reactive oxygen species. The aim of the present study was to investigate the superoxide release and cellular glutathione peroxidase (cGPx) activity in peripheral blood granulocytes and monocytes from children and adolescents with atopic asthma. Forty-four patients were selected and classified as having intermittent or persistent asthma (mild, moderate or severe). The spontaneous or phorbol myristate acetate (PMA, 30 nM)-induced superoxide release by granulocytes and monocytes was determined at 0, 5, 15, and 25 min. cGPx activity was assayed spectrophotometrically. The spontaneous superoxide release by granulocytes from patients with mild (N = 15), moderate (N = 12) or severe (N = 6) asthma was higher at 25 min compared to healthy individuals (N = 28, P < 0.05, Duncan test). The PMA-induced superoxide release by granulocytes from patients with moderate (N = 12) or severe (N = 6) asthma was higher at 15 and 25 min compared to healthy individuals (N = 28, P < 0.05 in both times of incubation, Duncan test). The spontaneous or PMA-induced superoxide release by monocytes from asthmatic patients was similar to healthy individuals (P > 0.05 in all times of incubation, Duncan test). cGPx activity of granulocytes and monocytes from patients with persistent asthma (N = 20) was also similar to healthy individuals (N = 10, P > 0.05, Kruskal-Wallis test). We conclude that, under specific circumstances, granulocytes from children with persistent asthma present a higher respiratory burst activity compared to healthy individuals. These findings indicate a risk of oxidative stress, phagocyte auto-oxidation, and the subsequent release of intracellular toxic oxidants and enzymes, leading to additional inflammation and lung damage in asthmatic children.

Effect of DEHP [di-(2-ethyl hexyl) phthalate] on lipid peroxidation in liver in rats and in primary cultures of rat hepatocytes.

The effect of DEHP [di-(2-ethly hexyl) phthalate] on lipid peroxidation in the liver in rats and in primary cultures of rat hepatocytes incubated with it was studied. The doses of DEHP used in this study corresponded to the low levels of this substance leaching into blood stored in DEHP plasticised PVC bags. Increased activity of superoxide dismutase (SOD) and catalase, increased concentration of malondialdehyde (MDA) and conjugated dienes and decrease in the concentration of glutathione and vitamin E have been observed in the liver of rats administered DEHP. Primary cultures of rat hepatocytes incubated with DEHP also showed increase in the activity of these enzymes, increase in the concentration of MDA and decrease in vitamin E. These results indicate that DEHP promotes lipid peroxidation. Incorporation of vitamin E along with DEHP into the culture medium containing hepatocytes counteracted these effects.

Action of the 4-nitro-2-phenoximethanesulphonanilide (nimesulide) on neutrophil chemotaxis and superoxide production

O 4-nitro-2-fenoximetanosulfonanilide (nimesulide), antiinflamatório näo esteróide, vem sendo utilizado no tratamento de quadros inflamatórios devido à sua atividade farmacológica nestes mecanismos de resposta as injúrias flogísticas. O objetivo do presente trabalho é estudar a açäo deste fármaco sobre duas importantes funçöes neutrófílicas: quimiotaxia e produçäo de ânion superóxido após pré incubaçäo com doses crescentes da droga (0,1; 0,3 e 0,5mM). Os resultados obtidos demonstram que a pré-incubaçäo de PMNs coletados do sangue periférico de indivíduos normais com o 4-nitro-2-fenoximetanosulfonanilide reduz significativamente a produçäo de superóxido por estimulaçäo de PMN com acetato de forbol miristato (PMA 50 ng/ml). formil metionil leucil fenilalanina (FMLP10-**7M e Zimozan opsonizado (1mg/ml). Adicionalmente o fármaco se mostrou igualmente efetivo em reduzir a habilidade quimiotáxica de PMNS, frente ao fator C5a(2 por cento de soro ativado por zimozan). FMLP (10-**5M) e leucotrieno B4(3x10-10**7M). Os resultados obtidos sugerem que além da sua atividade a nível do metabolismo do ácido araquidônico, o 4-nitro-2-fenoximetanosulfonanilide possa interferir diretamente sobre a funçäo neutrofílica, o que provavelmente contribui para sua açäo antiinflamatória

Xanthine oxidase, superoxide dismutase, catalase and lipid peroxidation in Mastomys natalensis: effect of Dipetalonema viteae infection.

Status of xanthine oxidase, superoxide dismutase, catalase and lipid peroxidation, the enzymes metabolizing reactive oxygen intermediates in liver, lungs and spleen of M. natalensis during D. viteae infection was investigated. Xanthine oxidase and lipid peroxidation exhibited stimulation, while superoxide dismutase and catalase showed depression in liver and spleen of the infected animals. The filarial infection therefore appears to create O2 toxicity in these tissues. Lungs, on the other hand was found safe as it possessed elevated xanthine oxidase, superoxide dismutase and catalase. Lipid peroxidation in lungs operated below the control level. The impact of these changes in the establishment and development of the infection has been discussed.