RESUMEN
BACKGROUND: We assessed the effect of hemodialysis (HD) and chronic kidney disease (CKD) on the serum levels of metalloproteinase-2 (MMP-2), MMP-9 and metalloproteinase tissue inhibitors (TIMP-1) and TIMP-2. METHODS: 18 patients on regular HD treatment with low-flux, cuprophane membrane, 15 non-dialyzed patients with CKD and 15 healthy controls were sampled. The serum MMP and TIMP concentrations were determined by ELISA assays. RESULTS: MMP-9, TIMP-1, and TIMP-2 serum levels were significantly decreased in HD patients to 32.7 ± 20.1 ng/ml, 178.8 ± 73.0 ng/ml, and 103.4 ± 55.3 ng/ml compared with 482.3 ± 139.5, 367.6 ± 75.5 ng/ml, and 299.7 ± 63.2 ng/ml in patients with CKD and 594.6 ± 154.7 ng/ml, 354.5 ± 81.2 ng/ml, and 272.4 ± 91.8 ng/ml in healthy controls, respectively, (P < 0.001 vs. HD patients). MMP-2 was lower in patients with CKD: 405.6 ± 106.1 ng/ml compared with 516.9 ± 81.7 ng/ml in controls (P = 0.02). The MMP-2/TIMP-2 ratio was increased in HD patients compared with both patients with CKD and controls. In the course of an HD session, MMP-2 and TIMP-1 serum levels were significantly decreased from pre-HD 570.0 ± 256.5 and 178.8 ± 66.9 ng/ml to post-HD 492.6 ± 212.5 and 144.6 ± 44.2 ng/ml (P = 0.004 and 0.013, respectively). However, the MMP-9/TIMP-1 ratio increased from pre-HD 0.15 (2.19) (median, range) to 0.23 (0.33) after a HD session (P = 0.03). CRP was positively correlated with MMP-9 and MMP-9/TIMP-1 ratio in HD patients and patients with CKD (r = 0.67; P = 0.03). CONCLUSIONS: The MMP-9/TIMP-1 ratio increased during HD sessions, although their absolute levels were lowered. This change may represent a chronic state of enhanced fibrosis in patients undergoing HD.
Asunto(s)
Metaloproteinasa 2 de la Matriz/sangre , Metaloproteinasa 9 de la Matriz/sangre , Diálisis Renal , Insuficiencia Renal Crónica/sangre , Insuficiencia Renal Crónica/terapia , Inhibidor Tisular de Metaloproteinasa-1/sangre , Inhibidor Tisular de Metaloproteinasa-2/sangre , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Renal replacement therapy (RRT) may differentially affect systemic generation of reactive oxygen species and depletion of antioxidant pools of low molecular weight molecules and proteins. This study was designed to assess the magnitude of the impairment of serum total antioxidant capacity (TAC) in relation to different RRT modalities. The study included patients on continuous ambulatory peritoneal dialysis (CAPD, N = 21), hemodialysis (HD, N = 21), hemodiafiltration (HDF, N = 20), and healthy controls (N = 33). TAC was assessed by the ferric reducing ability of plasma (FRAP) and with the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) assay. In CAPD patients, predialysis FRAP and DPPH were increased: 1.46 mM and 10.5% vs. control 1.19 mM and 7.2%, respectively (P < 0.001 in each). In HD and HDF patients, the FRAP and DPPH were significantly increased before and lowered after the RRT session (P < 0.05) if compared with healthy controls. During an HD session, FRAP was decreased from pre-HD 1.71 +/- 0.29 mM to post-HD 0.85 +/- 0.20 mM (P = 0.0001). The decrease of FRAP was lower during HDF (P < 0.05 vs. HD), it decreased from pre-HDF 1.41 +/- 0.43 mM to post-HDF 0.87 +/- 0.23 mM (P = 0.0001 vs. pre-HDF). The HD session decreased DPPH from the pre-HD median 10.3%, interquartile range (IR) 9.3-12.0% to post-HD 2.6% IR 2.3-3.1% (P < 0.0001). The adjustment of either urate or bilirubin up to pre-HD levels did not restore lowered post-HD levels of TAC. TAC remains preserved in CAPD, whereas the robust depletion of TAC, lower after HDF than HD sessions, cannot be attributed solely to the washout of dialyzable compounds.
Asunto(s)
Antioxidantes/metabolismo , Hemodiafiltración/efectos adversos , Diálisis Peritoneal Ambulatoria Continua/efectos adversos , Diálisis Renal/efectos adversos , Adulto , Anciano , Compuestos de Bifenilo , Estudios de Casos y Controles , Femenino , Compuestos Férricos/química , Hemodiafiltración/métodos , Humanos , Masculino , Persona de Mediana Edad , Diálisis Peritoneal Ambulatoria Continua/métodos , Picratos , Especies Reactivas de Oxígeno/metabolismo , Diálisis Renal/métodosRESUMEN
PURPOSE: This study was designed to assess exhaled hydrogen peroxide (H(2)O(2)), blood serum antioxidant capacity, and tumor necrosis factor-alpha (TNFalpha) in primary breast cancer (PBC). METHODS: The study included 34 consecutive, non-smoking PBC patients (aged 62.5 +/- 13.5 at surgery) prior to the treatments, qualified for modified radical mastectomy and not undergoing any adjuvant systemic therapy, and 33 healthy controls. The post surgery pathological assessment included tissue expression of estrogen (ER) and progesterone (PR) receptors, and epidermal growth factor receptor type 2 (HER-2/neu). Exhaled H(2)O(2) was determined fluorometrically in the exhaled breath condensate (EBC). Blood serum antioxidant capacity and TNFalpha levels were assessed with ferric reducing ability of plasma (FRAP) and ELISA immunoassay, respectively. RESULTS: In PBC patients, 10 ER, 11 PR, and 9 HER-2/neu positive tumors were identified and HER-2/neu score was 2+ in 20% of all tumors. Median (Me) H(2)O(2) was increased up to 0.44 microM (interquartile range IR: 0.20-1.25 microM) compared with healthy control of 0.36 microM (IR: 0.12-0.48 microM; p < 0.05). The H(2)O(2) concentration in EBC was significantly correlated (tau = 0.27; p = 0.03) and increased in cases with nodal metastases (n = 12; p = 0.04). Serum TNFalpha was increased up to 51.7 +/- 21.0 pg/ml compared with controls 17.2 +/- 3.65 pg/ml (p < 0.05). FRAP was increased to 1.41 +/- 0.37 mM Fe(2+) compared with control 1.19 +/- 0.17 mM Fe(2+); (p = 0.006). CONCLUSIONS: This is the first study to demonstrate increased H(2)O(2) in exhaled breath condensate in patients with localized breast malignancy and its relation with clinical severity.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Neoplasias de la Mama/metabolismo , Pruebas Respiratorias , Peróxido de Hidrógeno/metabolismo , Adulto , Anciano , Antioxidantes/metabolismo , Axila , Neoplasias de la Mama/patología , Estudios de Casos y Controles , FMN Reductasa/sangre , Femenino , Humanos , Inmunohistoquímica , Metástasis Linfática , Persona de Mediana Edad , Plasma/enzimología , Receptor ErbB-2/metabolismo , Factor de Necrosis Tumoral alfa/sangreRESUMEN
BACKGROUND: The regulation of mesangial extracellular matrix (ECM) turnover engages a number of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). High glucose concentration affects ECM degradation and the activities of MMPs and TIMPs. ECM accumulation is involved in the pathogenesis of diabetic nephropathy. METHODS: Serum MMP-9, MMP-2, TIMP-2 and TIMP-1 were measured with ELISA in patients with either chronic renal failure (CRF, n=20), type 2 diabetes mellitus (DM2, n=16) or diabetic nephropathy (DM2+CRF, n=14), and healthy controls (n=20). RESULTS: Diabetic nephropathy was related with profound decrease of serum TIMP-2 (122.2 +/- 47.2 vs. 263.0 +/- 89.2 ng/mL), TIMP-1 (242.5 +/- 96.9 vs. 347.4 +/- 87.2 ng/mL) and MMP-2 (385.4 +/- 42.6 vs. 517.2 +/- 75.4 ng/mL) (p<0.001). Both TIMP-1 and TIMP-2 were reduced in diabetic nephropathy in comparison with either diabetes alone (p<0.01 and p<0.001; respectively) or CRF alone (p<0.001 for both). An approximately 2-fold increase of MMP-9/TIMP-1 and MMP-2/TIMP-2 ratio was found in diabetic nephropathy when compared with diabetes with normal renal function (p<0.01). Further, in DM2 patients, TIMP-2 was decreased when compared with CRF alone (219.2 +/- 71.8 vs. 296.8 +/- 58.4 ng/mL). MMP-2 was lowered in both groups of DM2 and CRF patients (413.8 +/- 59.0 ng/mL and 409.7 +/- 93.1 ng/mL, vs. normal control value of 517.2 +/- 75.4 ng/mL; p<0.001). CONCLUSIONS: These data indicate that circulating TIMP-1, TIMP-2 and MMP-2 are decreased in patients with diabetic nephropathy when compared with either CRF or diabetes.
Asunto(s)
Nefropatías Diabéticas/diagnóstico , Metaloproteinasa 2 de la Matriz/sangre , Metaloproteinasa 9 de la Matriz/sangre , Inhibidor Tisular de Metaloproteinasa-1/sangre , Inhibidor Tisular de Metaloproteinasa-2/sangre , Anciano , Diabetes Mellitus Tipo 2/complicaciones , Femenino , Humanos , Fallo Renal Crónico/diagnóstico , Fallo Renal Crónico/etiología , Masculino , Persona de Mediana Edad , Factor de Necrosis Tumoral alfa/sangreRESUMEN
BACKGROUND: The oxidative burden in the airways is a hallmark of chronic obstructive pulmonary disease (COPD). AIMS: This prospective, cross-over, placebo (PL)-controlled study was designed to investigate the effect of N-acetyl-l-cysteine (NAC) on hydrogen peroxide (H(2)O(2)), nitrites and nitrates (NO(2)(-)+NO(3)(-)), and thiol (RSH) concentrations in exhaled breath condensate (EBC) in stable COPD patients (n=19, aged 52.6+/-15.6 years, 10 females, mean FEV(1) 95.2+/-23.8%, FEV(1)/FVC 69.1+/-11.4%). METHODS: H(2)O(2), NO(2)(-)+NO(3)(-) and RSH concentrations in EBC were determined with homovanillic acid, NADPH-nitrite reductase assays and Ellman's reaction, respectively. RESULTS: Thirty minutes after nebulization, H(2)O(2) concentration increased if levels after NAC (0.45+/-0.25microM) and PL (0.17+/-0.17microM) were compared in COPD patients (p=0.002). This increased H(2)O(2) level in EBC was no longer observed either after 90min: 0.16+/-0.09microM (PL 0.17+/-0.15microM) or 3h: 0.12+/-0.07microM (PL 0.21+/-0.23microM) (p=0.5 and 0.2, respectively). The levels of NO(2)(-) and NO(3)(-) did not differ between NAC and PL. There was no significant difference in RSH levels between nebulized NAC and PL. After nebulized NAC, however, exhaled RSH increased from 1.42+/-1.69microM (0min) to 2.49+/-2.00microM (30min), and 1.71+/-1.83microM (180min) (p=0.009 and 0.03, respectively, compared with 0min). CONCLUSIONS: These data demonstrate that nebulized NAC transiently increases exhaled H(2)O(2) level, whereas it has no effect on other oxidative parameters.
Asunto(s)
Acetilcisteína/uso terapéutico , Volumen de Reserva Espiratoria/efectos de los fármacos , Peróxido de Hidrógeno/metabolismo , Enfermedad Pulmonar Obstructiva Crónica/tratamiento farmacológico , Acetilcisteína/administración & dosificación , Anciano , Análisis de Varianza , Pruebas Respiratorias/métodos , Estudios Cruzados , Método Doble Ciego , Espiración , Expectorantes/administración & dosificación , Expectorantes/uso terapéutico , Femenino , Flujo Espiratorio Forzado/efectos de los fármacos , Volumen Espiratorio Forzado/efectos de los fármacos , Humanos , Peróxido de Hidrógeno/análisis , Masculino , Persona de Mediana Edad , Nebulizadores y Vaporizadores , Nitratos/análisis , Nitratos/metabolismo , Nitritos/análisis , Nitritos/metabolismo , Estudios Prospectivos , Enfermedad Pulmonar Obstructiva Crónica/metabolismo , Enfermedad Pulmonar Obstructiva Crónica/fisiopatología , Compuestos de Sulfhidrilo/análisis , Compuestos de Sulfhidrilo/metabolismoRESUMEN
BACKGROUND: The study aimed to differentiate the effects of hemodialysis (HD) and chronic renal failure (CRF) on the levels of circulating tumor necrosis factor-alpha (TNF-alpha) and TNF-alpha receptors p55 and p75, soluble vascular cell adhesion molecule-1 (sVCAM-1), intercellular adhesion molecule-1 (sICAM-1), soluble endothelial-leukocyte adhesion molecule-1 (sE-selectin) and sP-selectin in 18 patients on regular HD treatment with cuprophane membrane in relation to 15 non-dialyzed CRF patients and 15 healthy controls. METHODS: The serum concentrations were determined with standard ELISA assays. RESULTS: Blood serum p75 and p55 were approximately tenfold increased in CRF (36.7 +/- 6.2 and 27.1 +/- 5.6 ng/ml) and HD patients (45.6 +/- 18.4 and 28.7 +/- 5.9 ng/ml) before the HD session (HD 0), during (HD 20) the session (45.7 +/- 18.4 and 28.5 +/- 7.3 ng/ml) and after (HD 240) the HD session (52.1 +/- 17.4 and 30.9 +/- 8.2 ng/ml) in comparison to control values (5.6 +/- 1.3 and 2.4 +/- 0.8 ng/ml, respectively) (p < 0.01). The highest increment of p75 at the end of HD session (HD 240) was also significantly higher than at preceding time points (HD 0 and 20) (p < 0.05). However, the remaining study parameters did not change during an HD session. Also, there were no relevant changes in TNF-alpha levels if (HD 0) 22.7 +/- 21.5 ng/ml and (HD 240) 21.1 +/- 18.9 ng/ml were compared. Chronic HD status was related to the increase of sVCAM-1 and sICAM-1 levels. Prior to HD, T0 sVCAM-1 and sICAM-1 concentrations were 2,180.4 +/- 761.8 and 567.3 +/- 218.8 ng/ml, during HD (T20): 2,172.7 +/- 759.2 and 602.3 +/- 379.9 ng/ml, and after HD (T240): 2,401.6 +/- 756.4 and 648.3 +/- 183.5 ng/ml, respectively (p < 0.05 vs. controls and CRF patients). sVCAM-1 and sICAM-1 serum levels (1,262.2 +/- 472.9 and 165.6 +/- 50.4 ng/ml) were similar in CRF patients and healthy controls (854.4 +/- 241.5 and 217.6 +/- 74.2 ng/ml, respectively). Even though serum sE- and sP-selectin in CRF patients did not differ from the control (39.8 +/- 21.3 vs. 42.1 +/- 18.9 ng/ml and 187.9 +/- 66.9 vs. 198.8 +/- 62.2 ng/ml, respectively), their levels were increased in HD patients up to 111.9 +/- 54.6 and 453.2 +/- 231.1 ng/ml in patients prior to HD, 118.7 +/- 66.2 and 350.8 +/- 114.8 ng/ml during the HD session and then 132.3 +/- 61.1 and 368.3 +/- 126.6 ng/ml, respectively, after its completion (p < 0.05 in comparison with CRF patients and controls). CONCLUSIONS: The increased circulating TNF-alpha receptors appear more associated with the uremic milieu than HD-related systemic inflammation, whereas increased soluble cellular adhesion molecules in patients undergoing bioincompatible HD may be related to the enhanced systemic inflammation specifically due to maintenance HD.
Asunto(s)
Moléculas de Adhesión Celular/sangre , Receptores Tipo II del Factor de Necrosis Tumoral/sangre , Receptores Tipo I de Factores de Necrosis Tumoral/sangre , Diálisis Renal , Receptores Señuelo del Factor de Necrosis Tumoral/sangre , Adulto , Anciano , Anciano de 80 o más Años , Materiales Biocompatibles/metabolismo , Moléculas de Adhesión Celular/biosíntesis , Femenino , Humanos , Masculino , Persona de Mediana Edad , Diálisis Renal/efectos adversos , Factor de Necrosis Tumoral alfa/biosíntesis , Factor de Necrosis Tumoral alfa/sangreRESUMEN
INTRODUCTION: The study aimed to assess reactive oxygen species generation and the expressions of some surface antigens on polymorphonuclear leukocytes (PMNs) in patients on regular hemodialysis (HD) treatment. MATERIALS AND METHODS: The respiratory burst of PMNs was determined with luminol-dependent chemiluminescence (CL) in resting cells and following N-formyl-methionyl-leucyl-phenylalanine (fMLP), phorbol 12-myristate 13-acetate (PMA), or opsonized zymosan (OZ) stimulation and expressed in arbitrary CL units times assay-time (aU x min). The expressions of CD11b/CD18, CD10, and CD13 receptors were determined with flow cytometry. RESULTS: Basal PMN CL was increased in HD patients to up to 1285 +/- 129 aU x min compared with 895 +/- 88 aU x min in healthy controls (p < 0.05). The CL of unprimed PMNs increased after fMLP stimulation from 3085 +/- 746 to 4529 +/- 808 aU x min, and after OZ stimulation from 12945 +/- 1296 to 14678 +/- 1355 aU x min. PMA-stimulated CL of PMNs was similar to control values. The oxidative burst in PMNs from HD patients and healthy controls was similar in response to TNF-alpha alone. The CL of TNF-alpha-primed PMNs in HD patients was significantly lower than CL measured in healthy controls (p < 0.05). The expressions of CD10 and CD13 metalloproteinase receptors were also increased (p < 0.05). Although CD11b expression was significantly increased at rest and after fMLP stimulation, the expression of another beta-integrin heterodimer compound, CD18, was not increased. CONCLUSIONS: These results provide evidence that TNF-alpha priming of PMNs is down-regulated in HD patients despite constitutive up-regulation of resting cytotoxicity and enhanced expression of adhesion and metalloproteinase receptors.
Asunto(s)
Antígenos CD13/biosíntesis , Moléculas de Adhesión Celular/biosíntesis , Neprilisina/biosíntesis , Neutrófilos/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Diálisis Renal , Factor de Necrosis Tumoral alfa/fisiología , Adulto , Antígeno CD11b , Antígenos CD18/biosíntesis , Femenino , Humanos , Masculino , Neutrófilos/inmunología , Estallido Respiratorio , Uremia/inmunología , Uremia/metabolismoRESUMEN
This study was designed to investigate the direction of redox reactions of spermine and spermidine in the presence of iron and copper. The redox activity of spermine and spermidine was assessed using a variety of methods, including their ability to: (1) reduce Fe(3+) to Fe(2+) ions; (2) protect deoxyribose from oxidation by Fe(2+)-ethylene diaminetetraacetic acid, Fe(3+)-ethylene diaminetetraacetic acid systems with and without H(2)O(2); (3) protect DNA from damage caused by Cu(2+)-H(2)O(2), and Fe(2+)-H(2)O(2) with and without ascorbic acid; (4) inhibit H(2)O(2)-peroxidase-induced luminol dependent chemiluminescence; (5) scavenge diphenyl-picryl-hydrazyl radical. Spermine and spermidine at concentration 1mM reduced 1.8+/-0.3 and 2.5+/-0.1 nmol of Fe(3+) ions during 20 min incubation. Both polyamines enhanced deoxyribose oxidation. The highest enhancement of 7.6-fold in deoxyribose degradation was found for combination of spermine with Fe(3+)-ethylene diaminetetraacetic acid. An 10mM spermine and spermidine decreased CuSO(4)-H(2)O(2)-ascorbic acid- and FeSO(4)-H(2)O(2)-ascorbic-induced DNA damage by 73+/-6, 69+/-4% and 90+/-5, 53+/-4%, respectively. They did not protect DNA from CuSO(4)-H(2)O(2) and FeSO(4)-H(2)O(2). Spermine apparently increased the CuSO(4)-H(2)O(2)-dependent injury to DNA. Polyamines attenuated H(2)O(2)-peroxidase-induced luminol dependent chemiluminescence. Total light emission from specimens containing 10mM spermine or spermidine was attenuated by 85.3+/-1.5 and 87+/-3.6%. During 20 min incubation 1mM spermine or spermidine decomposed 8.1+/-1.4 and 9.2+/-1.8% of diphenyl-picryl-hydrazyl radical. These results demonstrate that polyamines of well known anti-oxidant properties may act as pro-oxidants and enhance oxidative damage to DNA components in the presence of free iron ions and H(2)O(2).