Arginine deficiency augments inflammatory mediator production by airway epithelial cells in vitro

<p>Abstract</p> <p>Background</p> <p>Previously we showed that reduced availability of the essential amino acid tryptophan per se attenuates post-transcriptional control of interleukin (IL)-6 and IL-8 leading to hyperresponsive production of these inflammatory mediators by airway epithelial cells. Availability of the non-essential amino acid arginine in the inflamed airway mucosa of patients with asthma is reduced markedly, but it is not known whether this can also lead to an exaggerated production of IL-6 and IL-8.</p> <p>Methods</p> <p>IL-6 and IL-8 were determined by ELISA in culture supernatants of NCI-H292 airway epithelial-like cells and normal bronchial epithelial (NHBE) cells that were exposed to TNF-α, LPS or no stimulus, in medium with or without arginine. Arginine deficiency may also result from exposure to poly-L-arginine or major basic protein (MBP), which can block arginine uptake. Epithelial cells were exposed to these polycationic proteins and L-¹⁴C-arginine uptake was assessed as well as IL-6 and IL-8 production. To determine the mode of action, IL-6 and IL-8 mRNA profiles over time were assessed as were gene transcription and post-transcriptional mRNA degradation.</p> <p>Results</p> <p>For both NCI-H292 and NHBE cells, low arginine concentrations enhanced basal epithelial IL-6 and IL-8 production and synergized with TNF-α-induced IL-6 and IL-8 production. Poly-L-arginine enhanced the stimulus-induced IL-6 and IL-8 production, however, blocking arginine uptake and the enhanced IL-6 and IL-8 production appeared unrelated. The exaggerated IL-6 and IL-8 production due to arginine deficiency and to poly-L-arginine depend on a post-transcriptional and a transcriptional process, respectively.</p> <p>Conclusion</p> <p>We conclude that both reduced arginine availability per se and the presence of polycationic proteins may promote airway inflammation by enhanced pro-inflammatory mediator production in airway epithelial cells, but due to distinct mechanisms.</p

O papel do óxido nítrico na pressão anal esfincteriana de ratos submetidos à colite experimental The role of nitric oxide in sphincteric anal pressure of rats with experimental colitis

O óxido nítrico (NO) é um radical livre sintetizado endogenamente por várias células do nosso organismo. Apresenta um amplo espectro de ações fisiológicas, sendo as mais importantes o seu mecanismo de ação parácrino no relaxamento da musculatura lisa, sua atividade neurotransmissora em vários sistemas e seu envolvimento no processo inflamatório. O NO é sintetizado em diferentes tecidos através da conversão da L-arginina em L-citrulina pela ação da enzima óxido nítrico sintase (NOS). OBJETIVOS: Este estudo tem por objetivo demonstrar o envolvimento do óxido nítrico no processo intestinal inflamatório de ratos Wistar submetidos à colite experimental com ácido acético. MATERIAL E MÉTODOS: Foram utilizados 20 ratos machos Wistar, com peso entre 250 e 350 gramas divididos em dois grupos de 10 animais. Os animais do grupo em estudo foram submetidos à administração intracolônica, por enema, de uma solução de ácido acético diluído a 7% e com volume de 3 ml. O grupo controle recebeu apenas enema de solução salina. Foram avaliados os índices histológicos, a expressão da enzima óxido nítrico sintase (iNOS) e a pressão anal esfincteriana. RESULTADOS: Os índices histológicos apresentaram uma significativa elevação no grupo colite quando comparados ao grupo controle, tanto na avaliação macroscópica quanto na microscópica. A expressão da enzima iNOS também foi significativamente maior no grupo colite quando comparada ao grupo controle. A pressão anal esfincteriana foi significativamente mais baixa no grupo colite na comparação ao grupo controle. CONCLUSÃO: Os animais submetidos à colite experimental apresentam um aumento da expressão da enzima óxido nítrico sintase induzível (i-NOS). Este aumento, associado ao conseqüente aumento do nível de óxido nítrico, ocasiona uma diminuição dos níveis de pressão anal esfincteriana.<br>The nitric oxide (NO) is a free radical synthesized from some cells of our organism. It presents with an ample specter of physiological actions being the most important its mechanism of action in the relaxation of the smooth musculature, its neurotransmissor activity in some systems and its involvement in the inflammatory process. The NO is synthesized in different tissues by the conversion of the L-arginine in L-citruline with the action of the enzyme nitric oxide sintase(NOS). OBJECTIVES: the aim of this study is to demonstrate the involvement of nitric oxide in the inflammatory intestinal process of Wistar rats submitted to experimental colitis with ascetic acid. MATERIAL AND METHODS: 20 male Wistar rats had been used with weight between 250 and 350 g divided in two groups of 10 animals. The animals of the group in study had been submitted to intracolonic administration, by enema, of a solution with acid ascetic diluted to 7% - 3 ml. The control group received only enema with saline solution. The histological scores, the expression of the enzyme nitric oxide sintase (iNOS) and the sphincteric anal pressure had been evaluated. RESULTS: The histological scores had presented a significant rise in the group colitis when compared with the control group in the macroscopic as well as in the microscopical evaluation. The expression of the enzyme iNOS was also significantly higher in the colitis group when compared to the control group. The sphincteric anal pressure was significantly lower in the group colitis when compared to control group. CONCLUSION: The animals submitted to the experimental colitis presented an increase of the iNOS expression. This increase, associated with the consequent increase in nitric oxide level, causes a reduction of the sphincteric anal pressure levels

Regulation of Nitric Oxide Production by δ-Opioid Receptors during Glaucomatous Injury

To determine the roles of nitric oxide in glaucomatous injury and its regulation by δ-opioid-receptor activation, animals were treated with: 1) a selective inducible nitric oxide synthase (iNOS) inhibitor (aminoguanidine; AG; 25 mg/kg, i.p.); 2) δ-opioid-receptor agonist (SNC-121; 1 mg/kg, i.p.); or 3) with both drugs simultaneously for 7 days, once daily. The loss in retinal ganglion cell (RGC) numbers and their function in glaucomatous eyes were significantly improved in the presence of AG or SNC-121; however, we did not see any significant additive or synergistic effects when animals were treated with both drugs simultaneously. The levels of nitrate-nitrite were significantly increased in the glaucomatous retina when compared with normal retina (normal retina 86±9 vs. glaucomatous retina 174±10 mM/mg protein), which was reduced significantly when animals were treated either with SNC-121 (121±7 mM/mg protein; P<0.05) or AG (128±10 mM/mg protein; P<0.05). Additionally, SNC-121-mediated reduction in nitrate-nitrite levels was not only blocked by naltrindole (a δ-opioid-receptor antagonist), but naltrindole treatment potentiated the nitrate-nitrite production in glaucomatous retina (235±4 mM/mg protein; P<0.001). As expected, naltrindole treatment also fully-blocked SNC-121-mediated retina neuroprotection. The nitrotyrosine level in the glaucomatous retina was also increased, which was significantly reduced in the SNC-121-treated animals. Additionally, the expression level of iNOS was clearly increased over the control levels in the glaucomatous retina and optic nerves, which was also reduced by SNC-121 treatment. In conclusion, our data support the notion that nitric oxide plays a detrimental role during glaucomatous injury and inhibition of nitric oxide production provided RGC neuroprotection. Furthermore, δ-opioid receptor activation regulates the production of nitric oxide via inhibiting the activity of iNOS in the retina and optic nerve