RESUMO
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.
Assuntos
Humanos , Anti-Inflamatórios não Esteroides/farmacologia , Materiais Biocompatíveis/farmacologia , Cinamatos/farmacologia , Depsídeos/farmacologia , Soluções para Hemodiálise/farmacologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Inflamação/tratamento farmacológico , Análise de Variância , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Citocinas/análise , Citocinas/efeitos dos fármacos , Formazans , Soluções para Hemodiálise/química , Células Endoteliais da Veia Umbilical Humana/metabolismo , Immunoblotting , Inflamação/metabolismo , Lipopolissacarídeos , NF-kappa B/análise , Óxido Nítrico/análise , Fosforilação , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Sais de TetrazólioRESUMO
OBJECTIVE To search for the reasons of unqualified level of bacteria in exit-entrance fluid of hemodialyzer and draft the measures of adjustment to meet the evaluation criteria for exit-entrance fluid of hemodialyzer.METHODS According to MOH′s Hospital Infection Management Standards to detect level and species of bacteria.RESULTS The average numbers of bacteria in exit-entrance fluid of hemodialyzer decreased from 2152.95?826.45 CFU/ml before adjustment to 579?541.04 CFU/ml after adjustment,checking with chi square test,the entrance fluid of hemodialyzer was ?2 =15.92,P