RESUMEN
Chronic systemic inflammation and repetitive damage of vascular endothelia by incompatible dialysis system are probable causes of cardiovascular disease in patients on dialysis. The present study aimed to assess in vitro biocompatibility and anti-inflammatory effect of hemodialysis fluid supplemented with rosmarinic acid (RA) using human umbilical vein endothelial cells (HUVEC). HUVECs (5×106 cells/mL) were pre-exposed to 1 µg/mL of lipopolysaccharides (LPS) and incubated with RA-supplemented hemodialysis fluid (HDF). Cytotoxicity was assessed qualitatively by morphologic assessment and quantitatively by MTT assay. Expressions of proinflammatory mediators were assessed using quantitative real-time PCR and production of NO was quantified. Phosphorylation of AKT and nuclear localization of nuclear factor kappa B (NF-κB) were examined using western blotting. Exposure of HUVECs to RA-supplemented HDF had no influence on morphology and viability. Inhibition of proinflammatory mediator production in HUVECs by RA supplementation to HDF was significant in a dose-dependent manner. Exposure to RA-supplemented HDF resulted in a decrease in nitric oxide synthase expression and reduction of NO production in LPS-stimulated HUVECs. RA supplementation of HDF suppressed Akt activation in LPS-stimulated HUVECs. In addition, the level of cellular IκB was increased in parallel to a reduced nuclear translocation of NF-κB in LPS-induced endothelial cells. Our results suggest that RA-supplemented HDF is biocompatible and significantly suppressed inflammation induced in endothelial cells. In this respect, the use of HDF supplemented with RA could alleviate inflammation and improve long-term treatment of patients with renal failure on dialysis. Further clinical studies are required to confirm the effects.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Materiales Biocompatibles/farmacología , Cinamatos/farmacología , Depsidos/farmacología , Soluciones para Hemodiálisis/farmacología , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Inflamación/tratamiento farmacológico , Análisis de Varianza , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Citocinas/análisis , Citocinas/efectos de los fármacos , Formazáns , Soluciones para Hemodiálisis/química , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Immunoblotting , Inflamación/metabolismo , Lipopolisacáridos , FN-kappa B/análisis , Óxido Nítrico/análisis , Fosforilación , Reacción en Cadena en Tiempo Real de la Polimerasa , Reproducibilidad de los Resultados , Sales de Tetrazolio , Ácido RosmarínicoRESUMEN
Este trabalho mostra as vantagens de se determinar simultaneamente os metabólitos VMA, 5HIAA, HVA urinários. A quantificação destes metabólitos são muito importantes nos diagnósticos e tratamento de tumores que se originam na crista neural (VMA, HVA), e diagnóstico da síndrome e tumor carcinóide (5HIAA). A maioria das técnicas utilizadas, limitam-se a medir somente um ou dois setes metabólitos por análise, sendo que a ESA atualmente desenvolveu um procedimento direto, onde se obtém uma quantificação simultânea dos três metabólitos (VMA, 5HIAA, HVA) urinários usando o Sistema Coulochem Electrode Array CEAS
Asunto(s)
Humanos , Técnicas de Laboratorio Clínico , Técnicas y Procedimientos Diagnósticos , OrinaRESUMEN
We compared intermittent (8 hours/day) versus continuous (24 hours/day) isocaloric lipid infusion regimens in 28 neonates. The lipid dose was increased incrementally by 0.5 gm/kg/day to either 3 gm/kg/day or until fat contributed 40% of daily calories. Serum total triglycerides, free fatty acids, free fatty acids/albumin molar ratio, and total cholesterol levels were measured prior to the daily lipid infusion, at the end of the intermittent infusion, and at 8 hours during the continuous infusion. Neonates less than 32 weeks postconception had significant fluctuation of triglycerides, free fatty acids, and free fatty acids/albumin molar ratio during the intermittent regimen at all lipid doses, but not during the continuous regimen. Neonates greater than or equal to 32 weeks postconception had significant fluctuation of serum triglycerides, free fatty acids, and free fatty acids/albumin molar ratio during the intermittent regimen with a lipid dose greater than or equal to 2 gm/kg/day, but not during the continuous regimen at all lipid doses. Serum free fatty acids correlated closely with serum triglycerides during both regimens (r = 0.89, P less than 0.001). Serum total cholesterol rose with increasing lipid doses during both regimens (f = 8.16, P less than 0.05). We conclude that neonates less than 32 weeks postconception tolerate the continuous regimen better than the intermittent regimen at all lipid doses; neonates greater than or equal to 32 weeks postconception tolerate both regimens well at lipid dose less than 2 gm/kg/day, but tolerate a continuous regimen better with lipid dose greater than or equal to 2 gm/kg/day.