RESUMEN
BACKGROUND: Reactive oxygen species (ROS) play a pivotal role in neointima hyperplasia after balloon injury. Molecular hydrogen has emerged as a novel antioxidant and has been proven effective in treating many diseases. OBJECTIVES: We aimed to determine the mechanism by which hydrogen affects neointima formation. METHODS: We assessed the influence of a hydrogen-rich saline solution (HRSS) by daily injection in rats. Rats were euthanized to evaluate the neointima. ROS, malondialdehyde (MDA) and superoxide dismutase (SOD) and reduced glutathione (GSH), were detected in the injured artery. Macrophage infiltration and the production of inflammatory factors (i.e., IL-6, TNF-α and NF-κB) were also observed. The in vitro effects of hydrogen on vascular smooth muscle cell (VSMC) proliferation were also measured. RESULTS: HRSS decreased the neointima area significantly. The neointima/media ratio was also reduced by HRSS. There was a decline in the number of PCNA-positive cells in the intima treated with HRSS. Meanwhile, HRSS ameliorated the ROS and MDA levels and increased SOD, reduced GSH levels in the injured carotid. In addition, the levels of inflammatory factors, such as IL-6, TNF-α and NF-κB p65, were attenuated by HRSS. In vitro studies also confirmed the anti-proliferative capability of the hydrogen solution and ROS generation in VSMCs induced by PDGF-BB. CONCLUSION: HRSS may have a protective role in the prevention of neointima hyperplasia and restenosis after angioplasty. HRSS may partially exert its role by neutralizing the local ROS and suppressing the TNF-α/NF-κB pathway.
Asunto(s)
Angioplastia de Balón , Antioxidantes/farmacología , Arterias Carótidas/efectos de los fármacos , Traumatismos de las Arterias Carótidas/tratamiento farmacológico , Proliferación Celular/efectos de los fármacos , Hidrógeno/farmacología , Mediadores de Inflamación/metabolismo , FN-kappa B/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Cloruro de Sodio/farmacología , Factor de Necrosis Tumoral alfa/metabolismo , Túnica Íntima/efectos de los fármacos , Animales , Antioxidantes/administración & dosificación , Becaplermina , Arterias Carótidas/inmunología , Arterias Carótidas/metabolismo , Arterias Carótidas/patología , Traumatismos de las Arterias Carótidas/etiología , Traumatismos de las Arterias Carótidas/inmunología , Traumatismos de las Arterias Carótidas/metabolismo , Traumatismos de las Arterias Carótidas/patología , Células Cultivadas , Modelos Animales de Enfermedad , Glutatión/metabolismo , Hidrógeno/administración & dosificación , Hidrógeno/sangre , Inyecciones Intraperitoneales , Interleucina-6/metabolismo , Malondialdehído/metabolismo , Músculo Liso Vascular/efectos de los fármacos , Músculo Liso Vascular/inmunología , Músculo Liso Vascular/lesiones , Músculo Liso Vascular/metabolismo , Miocitos del Músculo Liso/efectos de los fármacos , Miocitos del Músculo Liso/inmunología , Miocitos del Músculo Liso/metabolismo , Proteínas Proto-Oncogénicas c-sis/metabolismo , Ratas , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos , Cloruro de Sodio/administración & dosificación , Cloruro de Sodio/sangre , Superóxido Dismutasa/metabolismo , Factores de Tiempo , Túnica Íntima/inmunología , Túnica Íntima/metabolismo , Túnica Íntima/patologíaRESUMEN
The enhancement of re-endothelialisation is a critical therapeutic option for repairing injured blood vessels. Endothelial progenitor cells (EPCs) are the major source of cells that participate in endothelium repair and contribute to re-endothelialisation by reducing neointima formation after vascular injury. The over-expression of the inhibitor of differentiation or DNA binding 1 (Id1) significantly improved EPC proliferation. This study aimed to investigate the effects of Id1 on the phosphatidylinositol-3-kinase (PI3K)/Akt/nuclear factor kappa B (NFκB)/survivin signalling pathway and its significance in promoting EPC proliferation in vitro. Spleen-derived EPCs were cultured as previously described. Id1 was presented at low levels in EPCs, and was rapidly up-regulated by stimulation with vascular endothelial growth factor. We demonstrated that transient transfection of Id1 into EPCs activated the PI3K/Akt/NFκB/survivin signalling pathway and promoted EPC proliferation. The proliferation of EPCs was extensively inhibited by silencing of endogenous Id1, and knockdown of Id1 expression led to suppression of PI3K/Akt/NFκB/survivin signalling pathway in EPCs. In addition, blockade by the PI3K-specific inhibitor LY294002, Akt inhibitor, the NFκB inhibitor BAY 11-7082, the survivin inhibitor Curcumin, or the survivin inhibitor YM155 reduced the effects of Id1 transfection. These results suggest that the Id1/PI3K/Akt/NFκB/survivin signalling pathway plays a critical role in EPC proliferation. The Id1/PI3K/Akt/NFκB/survivin signalling pathway may represent a novel therapeutic target in the prevention of restenosis after vascular injury.
Asunto(s)
Proliferación Celular , Células Endoteliales/enzimología , Proteínas Inhibidoras de la Apoptosis/metabolismo , Proteína 1 Inhibidora de la Diferenciación/metabolismo , FN-kappa B/metabolismo , Fosfatidilinositol 3-Quinasa/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal , Células Madre/enzimología , Transporte Activo de Núcleo Celular , Animales , Núcleo Celular/enzimología , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Proteína 61 Rica en Cisteína/metabolismo , Células Endoteliales/efectos de los fármacos , Proteínas Inhibidoras de la Apoptosis/antagonistas & inhibidores , Proteína 1 Inhibidora de la Diferenciación/genética , FN-kappa B/antagonistas & inhibidores , Inhibidores de las Quinasa Fosfoinosítidos-3 , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Proto-Oncogénicas c-akt/antagonistas & inhibidores , Interferencia de ARN , Ratas , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos , Bazo/citología , Bazo/enzimología , Células Madre/efectos de los fármacos , Survivin , Transfección , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
The aggregation and adhesion of bacterial cells is a serious disadvantage for electrophoretic separations of bacteria. In this study, lipid-based liquid crystalline nanoparticles were used as a pseudostationary phase to minimise the bacterial aggregation and adsorption to the inner walls of microchannels. Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus and Lactobacillus rhamnosus were selected as analytes and were separated by microchip electrophoresis (MCE) with laser-induced fluorescence (LIF) detection using 4.5 mM tris(hydroxymethyl) aminomethane (TRIS)-4.5 mM boric acid-0.1 mM ethylenediaminetetraacetate (EDTA) (TBE) containing poly(ethylene oxide) (PEO) and lipid-based nanoparticles as the running buffer. The mechanism of lipid-based nanoparticles affecting bacterial adhesion and aggregation was discussed and supported by zeta potential experiments. Under the optimal conditions, the three species of bacteria were identified with patterned peaks. This proposed MCE method using lipid-based nanoparticles as running buffer additives was also used to analyse a real yogurt sample, and valuable bacterial information was obtained by the electropherograms.
Asunto(s)
Técnicas de Tipificación Bacteriana/métodos , Diglicéridos/química , Electroforesis por Microchip , Nanopartículas/química , Fosfatidilcolinas/química , Electroforesis por Microchip/instrumentación , Electroforesis por Microchip/métodos , Análisis de los Alimentos , Concentración de Iones de Hidrógeno , Lactobacillus/química , Polisorbatos/química , Streptococcus thermophilus/química , Yogur/análisis , Yogur/microbiologíaRESUMEN
OBJECTIVE: To observe the feasibility of measuring multiple circulating cytokines by protein chips in patients with acute coronary syndrome (ACS). METHODS: Circulating cytokines were determined by protein chips in 50 control cases and 104 ACS cases of unstable angina pectoris (UA, n = 50) and acute myocardial infarction (AMI, n = 54) cases. RESULTS: The levels of serum MMP-9, sCD40L, CRP, IL-6, H-FABP, cTnI and plasma IL-8, NT-proBNP, sVCAM-1, ET-1 in ACS group were significantly higher than those in the control group (all P < 0.01). Serum H-FABP and cTnI were significantly higher in AMI patients than in UA patients (P < 0.01). Serum MMP-9, sCD40L, CRP, IL-6 and plasma sVCAM-1, ET-1 tended to be higher in AMI patients as compared to UA patients (all P > 0.05). Serum MMP-9, sCD40L, and H-FABP were positively correlated with the severity of angina(r = 0.653, r = 0.745, r = 0.933, P < 0.01, respectively). Serum MMP-9, sCD40L, and H-FABP increased in proportion to the levels of Braunwald angina classes in UA patients (angina class I < angina class II < angina class III, P < 0.01). CONCLUSIONS: Circulating cytokines were significantly increased in ACS patients and positively correlated to Braunwald angina classes in UA patients.