RESUMO
To take cancer survivorship research to the next level, it's important to gain insight in trajectories of changing patient-reported outcomes and impaired recovery after cancer. This is needed as the number of survivors is increasing and a large proportion is confronted with changing health after treatment. Mechanistic research can facilitate the development of personalized risk-stratified follow-up care and tailored interventions to promote healthy cancer survivorship. We describe how these trajectories can be studied by taking the recently extended Dutch population-based Patient Reported Outcomes Following Initial treatment and Long term Evaluation of Survivorship (PROFILES) registry as an example. PROFILES combines longitudinal assessment of patient-reported outcomes with novel, ambulatory and objective measures (eg, activity trackers, blood draws, hair samples, online food diaries, online cognitive tests, weighing scales, online symptoms assessment), and cancer registry and pharmacy databases. Furthermore, we discuss methods to optimize the use of a multidomain data collection-like return of individual results to participants, which may improve not only patient empowerment but also long-term cohort retention. Also, advanced statistical methods are needed to handle high-dimensional longitudinal data (with missing values) and provide insight into trajectories of changing patient-reported outcomes after cancer. Our coded data can be used by academic researchers around the world. Registries like PROFILES, which go beyond boundaries of disciplines and institutions, will contribute to better predictions of who will experience changes and why. This is needed to prevent and mitigate long-term and late effects of cancer treatment and to identify new interventions to promote health.
Assuntos
Sobreviventes de Câncer , Neoplasias , Promoção da Saúde , Humanos , Neoplasias/psicologia , Neoplasias/terapia , Medidas de Resultados Relatados pelo Paciente , Sistema de Registros , Sobreviventes/psicologiaRESUMO
Supplementation with nicotinamide adenine dinucleotide (NAD+) precursors including dietary nicotinamide has been found to boost tissue NAD+ levels and ameliorate oxidative stress-induced damage that contributes to aging and aging-related diseases. The association between dietary NAD+ precursors and patient-reported health-related outcomes in cancer survivors has not been investigated. This study aimed to determine associations of dietary nicotinamide intake with different patient-reported outcomes in colorectal cancer survivors, 2 to 10 years post-diagnosis. A total of 145 eligible participants were recruited into this cross-sectional study. Dietary nicotinamide intake level was calculated based on data from 7-day food diaries. Fatigue was assessed with the Checklist Individual Strength (CIS), which is a subscale of the cancer-specific European Organization for the Research and Treatment of Cancer Quality of Life Questionnaire-Core 30 (EORTC), and anxiety and depression were assessed with Hospital Anxiety and Depression Scale (HADS). Oxidative stress marker serum protein carbonyl contents and serum NAD+ levels were measured. A hierarchical linear regression model with confounder adjustment was performed to analyze the association of nicotinamide intake, serum protein carbonyl contents, and NAD+ levels with patient-reported outcomes. The median values of daily nicotinamide intake for male and female participants were 19.1 and 14.4 mg, respectively. Daily dietary nicotinamide intake was associated with a lower level of fatigue (ß: -14.85 (-28.14, -1.56)) and a lower level of anxiety and depression (ß: -4.69 (-8.55, -0.83)). Subgroup analyses by sex showed that a beneficial association between nicotinamide intake and patient-reported outcomes was mainly found in men. To conclude, our findings suggested that higher dietary NAD+ precursor nicotinamide intake was cross-sectionally associated with less patient-reported outcomes in CRC survivors.
Assuntos
Sobreviventes de Câncer , Neoplasias Colorretais/patologia , Dieta , Niacinamida/farmacologia , Medidas de Resultados Relatados pelo Paciente , Idoso , Ansiedade/complicações , Sobreviventes de Câncer/psicologia , Cognição , Neoplasias Colorretais/psicologia , Estudos Transversais , Depressão/complicações , Emoções , Fadiga/complicações , Feminino , Força da Mão , Humanos , MasculinoRESUMO
Vitamin B6 is a water-soluble vitamin that functions as a coenzyme in many reactions involved in amino acid, carbohydrates and lipid metabolism. Since 2014, >50 cases of sensory neuronal pain due to vitamin B6 supplementation were reported. Up to now, the mechanism of this toxicity is enigmatic and the contribution of the various B6 vitamers to this toxicity is largely unknown. In the present study, the neurotoxicity of the different forms of vitamin B6 is tested on SHSY5Y and CaCo-2 cells. Cells were exposed to pyridoxine, pyridoxamine, pyridoxal, pyridoxal-5-phosphate or pyridoxamine-5-phosphate for 24h, after which cell viability was measured using the MTT assay. The expression of Bax and caspase-8 was tested after the 24h exposure. The effect of the vitamers on two pyridoxal-5-phosphate dependent enzymes was also tested. Pyridoxine induced cell death in a concentration-dependent way in SHSY5Y cells. The other vitamers did not affect cell viability. Pyridoxine significantly increased the expression of Bax and caspase-8. Moreover, both pyridoxal-5-phosphate dependent enzymes were inhibited by pyridoxine. In conclusion, the present study indicates that the neuropathy observed after taking a relatively high dose of vitamin B6 supplements is due to pyridoxine. The inactive form pyridoxine competitively inhibits the active pyridoxal-5'-phosphate. Consequently, symptoms of vitamin B6 supplementation are similar to those of vitamin B6 deficiency.
Assuntos
Suplementos Nutricionais/toxicidade , Piridinas/toxicidade , Alanina Transaminase/metabolismo , Células CACO-2 , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Polineuropatias/induzido quimicamente , Tirosina Descarboxilase/metabolismo , Deficiência de Vitamina B 6 , Vitaminas/toxicidadeRESUMO
BACKGROUND: Accelerated aging has been proposed as a pathologic mechanism of various chronic diseases, including COPD. This concept has almost exclusively been approached by analyses of individual markers. We investigated whether COPD is associated with accelerated aging using a panel of markers representing various interconnected aspects of the aging process. METHODS: Lung function, leukocyte telomere length, lymphocyte gene expression of anti-aging (sirtuin 1, total klotho, and soluble klotho [Sklotho]), senescence (p16/21), and DNA repair (Ku70/80 and TERF2) proteins, and markers of systemic inflammation and oxidative stress were determined in 160 patients with COPD, 82 smoking subjects, and 38 never-smoking control subjects. RESULTS: Median levels for telomere length, Sklotho, Ku70, and sirtuin 1 gene expression were lower (respectively, 4.4, 4.6, and 4.7 kbp for telomere length; 74%, 82%, and 100% for Sklotho; 88%, 92%, and 100% for Ku70 and 70%, 92%, and 100% for sirtuin 1, all P < .05) in patients compared with the smoking and never-smoking control groups. P21 gene expression was higher in patients compared with smoking control subjects. Telomere length correlated with Ku70 gene expression (r = 0.15, P = .02). After correction for age, smoking history, systemic inflammation, and oxidative stress, telomere length and p21 were the only markers that remained independently associated with lung function. In separate groups, only telomere length remained associated with lung function parameters. CONCLUSIONS: Markers of the aging mechanism represent distinct molecular aspects of aging. Among them, different markers were altered in COPD, but only telomere length was consistently associated with lung function, and seems a useful marker for expressing accelerated aging in COPD.
Assuntos
Envelhecimento/fisiologia , Doença Pulmonar Obstrutiva Crônica/metabolismo , Homeostase do Telômero/fisiologia , Idoso , Biomarcadores/metabolismo , Estudos de Casos e Controles , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Reparo do DNA/fisiologia , Feminino , Glucuronidase/metabolismo , Humanos , Proteínas Klotho , Masculino , Pessoa de Meia-Idade , Estresse Oxidativo/fisiologia , Doença Pulmonar Obstrutiva Crônica/complicações , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Sirtuína 1/metabolismo , FumarRESUMO
Oxidative stress plays a major role in the pathophysiology of chronic inflammatory disease and it has also been linked to accelerated telomere shortening. Telomeres are specialized structures at the ends of linear chromosomes that protect these ends from degradation and fusion. Telomeres shorten with each cell division eventually leading to cellular senescence. Research has shown that poly(ADP-ribose) polymerase-1 (PARP-1) and subtelomeric methylation play a role in telomere stability. We hypothesized that PARP-1 plays a role in accelerated aging in chronic inflammatory diseases due to its role as coactivator of NF-κb and AP-1. Therefore we evaluated the effect of chronic PARP-1 inhibition (by fisetin and minocycline) in human fibroblasts (HF) cultured under normal conditions and under conditions of chronic oxidative stress, induced by tert-butyl hydroperoxide (t-BHP). Results showed that PARP-1 inhibition under normal culturing conditions accelerated the rate of telomere shortening. However, under conditions of chronic oxidative stress, PARP-1 inhibition did not show accelerated telomere shortening. We also observed a strong correlation between telomere length and subtelomeric methylation status of HF cells. We conclude that chronic PARP-1 inhibition appears to be beneficial in conditions of chronic oxidative stress but may be detrimental under relatively normal conditions.
Assuntos
Senescência Celular , Estresse Oxidativo , Poli(ADP-Ribose) Polimerases/metabolismo , Senescência Celular/efeitos dos fármacos , Cromossomos Humanos Par 2/metabolismo , Metilação de DNA/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Fibroblastos/enzimologia , Fibroblastos/patologia , Flavonoides/farmacologia , Flavonóis , Células HeLa , Humanos , Peróxido de Hidrogênio/farmacologia , Minociclina/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Poli(ADP-Ribose) Polimerase-1 , Poli Adenosina Difosfato Ribose/metabolismo , Telomerase/metabolismo , Telômero/metabolismo , Homeostase do Telômero/efeitos dos fármacos , terc-Butil Hidroperóxido/farmacologiaAssuntos
Envelhecimento/genética , Mortalidade , Fumar , Telômero/química , Feminino , Humanos , MasculinoRESUMO
BACKGROUND: Nuclear factor (NF)-κB activation and oxidative stress are physiologic responses of skeletal muscle to exercise but may be impaired in patients with COPD. Therefore, we investigated NF-κB activity and expression of NF-κB-regulated genes in muscle of patients with COPD and control subjects before and after exercise. METHODS: Quadriceps specimens were obtained before, immediately after, and 2 h after a submaximal cycle ergometry test from seven patients with COPD (50.6 ± 5.7 SEM FEV(1) of patients with COPD) and seven age-matched control subjects. NF-κB DNA-binding activity in muscle and peripheral blood mononuclear cells (PBMCs) was determined using electrophoretic mobility shift assay and enzyme-linked immunosorbent assay, respectively. mRNA expression and protein carbonylation were measured by real-time polymerase chain reaction and Western blot, respectively. RESULTS: In control subjects, IL-6, IκBα, tumor necrosis factor-α, IL-1ß, superoxide dismutase, thioredoxin, heme oxygenase 1, and heat shock protein-70 were upregulated in muscle after exercise, whereas in patients with COPD only IL-6 mRNA was increased. Exercise-induced antiapoptotic Bcl2 mRNA levels were attenuated in patients with COPD compared with control subjects. Basal muscle protein oxidation was higher in patients with COPD than in control subjects, but attenuated in response to exercise. No exercise-induced changes in NF-κB DNA-binding activity in muscle and PBMCs of either group were detected. CONCLUSIONS: Skeletal muscle of patients with COPD is characterized by an impaired response to exercise of NF-κB-regulated genes encoding inflammatory cytokines, antioxidants, stress proteins, and survival factors.
Assuntos
Teste de Esforço , Músculo Esquelético/metabolismo , NF-kappa B/metabolismo , Doença Pulmonar Obstrutiva Crônica/genética , Biópsia , Western Blotting , Estudos de Casos e Controles , Ensaio de Desvio de Mobilidade Eletroforética , Ensaio de Imunoadsorção Enzimática , Expressão Gênica , Proteínas de Choque Térmico HSP70/metabolismo , Heme Oxigenase-1/metabolismo , Humanos , Quinase I-kappa B/metabolismo , Interleucina-1/metabolismo , Interleucina-6/metabolismo , Masculino , Pessoa de Meia-Idade , Estresse Oxidativo , Doença Pulmonar Obstrutiva Crônica/metabolismo , RNA Mensageiro/metabolismo , Testes de Função Respiratória , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estatísticas não Paramétricas , Superóxido Dismutase/metabolismo , Tiorredoxinas/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Regulação para CimaRESUMO
Telomere shortening is a marker of aging and therefore telomere length might be related to disease progression and survival. To address these questions, we measured leukocyte telomere length (LTL) in male participants from the Zutphen Elderly Study. LTL was measured by quantitative polymerase chain reaction in 203 men: mean aged 78 years in 1993 and 75 surviving participants mean aged 83 years in 2000. During 7 years of follow-up, 105 men died. Cox proportional hazards models were used to estimate hazard ratios for all-cause and cause-specific mortality. We found that LTL declined with a mean of 40.2 bp/year, and LTL values measured in 1993 and 2000 correlated significantly (r = .51, p < .001). Longer telomeres at baseline were not predictive for all-cause mortality, cardiovascular mortality, or cancer mortality. These results suggest that LTL decreases with increasing age and that LTL is not related to mortality in men aged more than 70 years.
Assuntos
Mortalidade , Telômero , Idoso , Idoso de 80 Anos ou mais , Biomarcadores , Humanos , Leucócitos/metabolismo , Masculino , Modelos de Riscos Proporcionais , Estudos ProspectivosRESUMO
The nuclear enzyme poly(ADP-ribose) polymerse-1 (PARP-1) has previously been reported to play an important role in lipopolysaccharide (LPS)-induced pulmonary inflammation and is highly activated in COPD patients. In the present study, the anti-inflammatory efficacy of a previously identified poly(ADP-ribose) polymerase-1 (PARP-1) inhibiting caffeine metabolite, 1,7-dimethylxanthine, was both in vivo as well as ex vivo evaluated. Orally administered 1,7-dimethylxanthine significantly attenuated lung myeloperoxidase-levels, transcription of IL-6, TNF-alpha, MIP1alpha and MIP2 genes as well as PAR-polymer formation in a mouse model of intratracheally LPS-induced acute pulmonary inflammation. Serum amyloid P component and plasma IL-6 were also lowered in 1,7-dimethylxanthine treated mice, indicating a reduced systemic inflammatory response. In addition, at 24h after LPS administration anti-inflammatory effects of 1,7-dimethylxanthine appeared more pronounced than those of the orally administered PARP-1 inhibitor 3-aminobenzamide. In the second model, in blood of COPD-patients and healthy controls ex vivo pre-incubated with a physiological concentration of 1,7-dimethylxanthine (10microM), LPS-induced production of the cytokines IL-6 and TNF-alpha was significantly suppressed. 1,7-Dimethylxanthine exerts anti-inflammatory effects, both in vivo mouse as well as ex vivo human. These results suggest that the PARP-1 inhibiting caffeine metabolite 1,7-dimethylxanthine may have therapeutic potential in pulmonary inflammatory diseases such as COPD.
Assuntos
Anti-Inflamatórios/farmacologia , Inibidores Enzimáticos/farmacologia , Pneumonia/tratamento farmacológico , Teofilina/farmacologia , Administração Oral , Idoso , Animais , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/uso terapêutico , Benzamidas/farmacologia , Estudos de Casos e Controles , Citocinas/sangue , Citocinas/metabolismo , Inibidores Enzimáticos/administração & dosagem , Inibidores Enzimáticos/uso terapêutico , Feminino , Humanos , Interleucina-6/sangue , Lipopolissacarídeos/farmacologia , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Masculino , Camundongos , Pessoa de Meia-Idade , Peroxidase/metabolismo , Pneumonia/metabolismo , Pneumonia/patologia , Poli(ADP-Ribose) Polimerase-1 , Poli Adenosina Difosfato Ribose/biossíntese , Inibidores de Poli(ADP-Ribose) Polimerases , Poli(ADP-Ribose) Polimerases/metabolismo , Doença Pulmonar Obstrutiva Crônica/sangue , Doença Pulmonar Obstrutiva Crônica/induzido quimicamente , Doença Pulmonar Obstrutiva Crônica/metabolismo , Doença Pulmonar Obstrutiva Crônica/patologia , Teofilina/administração & dosagem , Teofilina/uso terapêutico , Transcrição Gênica/efeitos dos fármacosRESUMO
BACKGROUND: Our aim for this study was to disentangle the contribution of muscular vs pulmonary oxidative stress during endurance exercise in patients with COPD. METHODS: Fifteen COPD patients and 10 healthy age-matched control subjects performed a continuously submaximal single-leg ergometer test (40% of peak workload) for 20 min or until they stopped (muscle endurance [Tlim]). Venous blood, urine samples, and exhaled breath condensate were sampled before, immediately after, and 2 h after exercise. RESULTS: Tlim was lower in COPD patients than in control subjects (p < 0.01). No exercise-induced systemic inflammation (ie, no raised levels of interleukin-6 or tumor necrosis factor-alpha) was found in the groups. Urinary malondialdehyde and uric acid levels (p < 0.05) were increased in COPD patients, whereas erythrocyte oxidized glutathione/reduced glutathione levels tended to be increased in COPD patients compared with control subjects after exercise (p = 0.08). Despite the relatively low cardioventilatory response to this localized muscle exercise, hydrogen peroxide levels in breath condensate significantly increased in COPD patients (p < 0.01). Nuclear factor kappaB DNA-binding activity of p50 in peripheral blood monocytes was elevated after exercise in both COPD patients (p < 0.01) and control subjects (p < 0.05), whereas p65 protein levels were not altered. CONCLUSION: COPD patients showed increased pulmonary and systemic oxidative stress after localized leg muscle exercise compared with healthy control subjects, without evidence of increased levels of systemic inflammation.
Assuntos
Exercício Físico/fisiologia , Perna (Membro)/fisiologia , Pulmão/fisiopatologia , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Índice de Massa Corporal , Ergometria , Tolerância ao Exercício , Fadiga/fisiopatologia , Feminino , Volume Expiratório Forçado , Humanos , Inflamação/fisiopatologia , Masculino , Pessoa de Meia-Idade , Estresse Oxidativo/fisiologia , Resistência Física , Valores de ReferênciaRESUMO
BACKGROUND: Impaired skeletal muscle function contributes to exercise intolerance in patients with COPD. Exercise-induced oxidative stress may initiate or accelerate impaired muscle function. Dichloroacetate (DCA) activates muscle pyruvate dehydrogenase complex (PDC) at rest, reducing inertia in mitochondrial energy delivery at the onset of exercise and thereby diminishing anaerobic energy production. This study aimed to determine whether DCA infusion also may reduce exercise-induced systemic oxidative stress and inflammatory response in patients with COPD. METHODS: A randomized, double-blind crossover design was used in which 13 patients with COPD performed maximal cycle exercise after an IV infusion of DCA (50 mg/kg body mass) or saline solution (placebo). Venous blood was sampled before exercise, and immediately, 30 min, and 2 h after exercise. Urine samples were obtained before and 2 h after exercise. RESULTS: Peak workload improved significantly after DCA infusion compared to placebo (10%; p < 0.01). Urinary uric acid levels after exercise were significantly lower in the DCA condition than in the placebo condition, whereas no significant difference was observed for urinary malondialdehyde levels. Oxidized glutathione (GSSG) levels were significantly increased 2 h after exercise in the placebo condition (p < 0.02) but not after DCA infusion. No changes in reduced glutathione (GSH), GSSG/GSH ratio, and superoxide dismutase activity were observed. Plasma interleukin (IL)-6 levels significantly increased 2 h after exercise only in the DCA condition (p < 0.01). CONCLUSIONS: This study shows that improved performance after a pharmacologic intervention known to activate PDC was accompanied by an enhanced IL-6 response and a limited reduction in exercise-induced systemic oxidative stress.
Assuntos
Ácido Dicloroacético/farmacologia , Exercício Físico/fisiologia , Estresse Oxidativo/efeitos dos fármacos , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Idoso , Biomarcadores/sangue , Biomarcadores/urina , Composição Corporal , Estudos Cross-Over , Ácido Dicloroacético/administração & dosagem , Método Duplo-Cego , Teste de Esforço , Feminino , Glutationa/metabolismo , Humanos , Infusões Intravenosas , Interleucina-6/sangue , Masculino , Malondialdeído/urina , Placebos , Espirometria , Ácido Úrico/urinaRESUMO
Recently, we identified several flavonoids as inhibitors of the nuclear enzyme poly(ADP-ribose) polymerase (PARP)-1 in vitro and in vivo. PARP-1 is recognized as coactivator of nuclear factor-kappaB and plays a role in the pathophysiology of diseases with low-grade systemic inflammation, such as chronic obstructive pulmonary disease (COPD) and type 2 diabetes (T2D). In this study, we assessed the antiinflammatory effects of flavonoids with varying PARP-1-inhibiting effects in whole blood from male patients with COPD or T2D and healthy men. A total of 10 COPD, 10 T2D patients, and 10 healthy volunteers matched for age and BMI were recruited. Blood from each participant was exposed to 1 microg/L lipopolysaccharide (LPS) over 16 h with or without preincubation with 10 micromol/L of flavone, fisetin, morin, or tricetin. Concentrations of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, -8, and -10 were measured in the supernatant. Preincubation with fisetin and tricetin strongly attenuated LPS-induced increases in concentrations of TNFalpha in blood from COPD patients [mean (+/- SEM): -41 +/- 4% (fisetin) and -31 +/- 4% (tricetin); P < 0.001] and IL-6 in blood from T2D patients [-31 +/- 5% (fisetin) and -29 +/- 6% (tricetin); P < or = 0.001]. Moreover, LPS-induced changes in TNFalpha and IL-6 concentrations were positively correlated with the extent of reduction by fisetin and tricetin. The PARP-1-inhibiting flavonoids fisetin and tricetin were able to attenuate LPS-induced cytokine release from leukocytes of patients with chronic systemic inflammation, indicating a potential application as nutraceutical agents for these patient groups.
Assuntos
Citocinas/sangue , Diabetes Mellitus Tipo 2/sangue , Inibidores Enzimáticos/farmacologia , Flavonoides/farmacologia , Inibidores de Poli(ADP-Ribose) Polimerases , Doença Pulmonar Obstrutiva Crônica/sangue , Idoso , Cromonas/farmacologia , Flavonóis , Humanos , Interleucina-1/sangue , Interleucina-10/sangue , Interleucina-8/sangue , Lipopolissacarídeos/farmacologia , Masculino , Pessoa de Meia-Idade , Poli(ADP-Ribose) Polimerase-1 , Fator de Necrose Tumoral alfa/sangueRESUMO
In the present study, the anti-inflammatory effects of the flavonoids flavone, fisetin and tricetin were evaluated in a mouse model of LPS-induced acute pulmonary inflammation. The flavonoid fisetin significantly reduced lung myeloperoxidase-levels and gene-expression of inflammatory mediators such as IL-6, TNF-alpha, IL-1beta, MIP-1alpha and MIP-2. The LPS-induced gene transcription of HO-1 and SOD2 was also significantly reduced by fisetin. Overall, the anti-inflammatory effects of fisetin in this in vivo model were much more pronounced as compared to the observed effects of flavone or tricetin and the anti-inflammatory glucocorticoid dexamethasone. The results of this study indicate that flavonoids such as fisetin might be potential candidates as pharmaceuticals or nutraceuticals in the treatment of pulmonary inflammatory diseases.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Cromonas/uso terapêutico , Flavonoides/uso terapêutico , Pneumonia/tratamento farmacológico , Animais , Flavonas , Flavonóis , Expressão Gênica , Heme Oxigenase-1/genética , Mediadores da Inflamação/antagonistas & inibidores , Mediadores da Inflamação/metabolismo , Interleucina-6/sangue , Lipopolissacarídeos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Infiltração de Neutrófilos , Peroxidase/antagonistas & inibidores , Pneumonia/imunologia , Poli(ADP-Ribose) Polimerase-1 , Poli(ADP-Ribose) Polimerases/metabolismo , Superóxido Dismutase/genéticaRESUMO
Chronic oxidative stress and systemic inflammation contribute to the pathology of several chronic diseases, one among which is chronic obstructive pulmonary disease (COPD). In addition, increased oxidative stress and inflammation have been observed to be negatively associated with telomere length (TL). Our aim was to investigate the TL in COPD patients in relation to pulmonary and extrapulmonary disease severity. Furthermore, based on experimental evidence suggesting the effects of oxidative stress on telomere shortening, we studied the association of TL with the antioxidant enzyme superoxide dismutase (SOD). One hundred and two COPD patients with moderate to severe COPD were studied and compared with 19 healthy age-matched controls. Patients were characterized by elevated levels of inflammatory markers (CRP, sTNF-receptors) and lower SOD-activity than the controls (p<0.001), irrespective of the SOD genotype. TL was negatively associated with age (p<0.01) and was significantly shorter in COPD patients than controls (p<0.05). Within the patient group age-adjusted TL variability could not be explained by lung function and smoking history but a modest association was found with the percentage of fat mass (p<0.05). These data provide evidence for a relationship between a disturbed oxidant/antioxidant balance and telomere shortening and indicate that preservation of fat mass may be protective in delaying telomere shortening in COPD patients.
Assuntos
Catalase/metabolismo , Leucócitos/metabolismo , Estresse Oxidativo/genética , Doença Pulmonar Obstrutiva Crônica/genética , Superóxido Dismutase/metabolismo , Telômero , Composição Corporal/fisiologia , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Estresse Oxidativo/fisiologia , Reação em Cadeia da Polimerase , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Testes de Função Respiratória , Fumar/metabolismoRESUMO
The nuclear enzyme poly(ADP-ribose) polymerase-1 (PARP-1), which was initially known to be highly activated by oxidative stress-induced DNA strand breaks, has been shown to be involved in the pathophysiology of acute and chronic inflammatory diseases. PARP-1 deficiency in mice led to the discovery of its coactivating function in the nuclear factor-kappa B-mediated gene expression and in addition, pharmaceutical inhibition of PARP-1 was shown to reduce the production of inflammatory mediators. In this study, the in vitro PARP-1-inhibiting effect of various flavonoids was investigated. The flavonoids myricetin, tricetin, gossypetin, delphinidin, quercetin, and fisetin were identified as significant inhibitors of the purified enzyme. Further evaluation of these compounds in N-methyl-N'-nitro-N-nitrosoguanidine-treated human pulmonary epithelial cells showed that the formation of the poly(ADP-ribose) polymers, as well as the decreased NAD(+) levels, was reduced by quercetin, fisetin, and tricetin. Finally, IL-8 production of LPS-stimulated human pulmonary epithelial cells could be significantly reduced by these flavonoids. The results of this study indicate that specific flavonoids have PARP-1-inhibiting activity in addition to the earlier described antioxidant effects. PARP-1 inhibition and preservation of cellular NAD(+) and energy production could play a role in the antiinflammatory activity of these specific flavonoids. In addition, these results indicate additional mechanisms by which flavonoids can exert antiinflammatory activity. Furthermore, these results indicate possibilities to use food-derived flavonoids in the treatment of chronic inflammatory diseases.
Assuntos
Flavonas/farmacologia , Flavonóis/farmacologia , Inibidores de Poli(ADP-Ribose) Polimerases , Mucosa Respiratória/citologia , Mucosa Respiratória/efeitos dos fármacos , Linhagem Celular Tumoral , Dieta , Humanos , Interleucina-8/metabolismo , Pulmão/citologia , Pulmão/metabolismo , Metilnitronitrosoguanidina/farmacologia , NAD/metabolismo , NF-kappa B/metabolismo , Poli(ADP-Ribose) Polimerase-1 , Mucosa Respiratória/metabolismoRESUMO
The nuclear enzyme poly(ADP-ribose) polymerase-1 (PARP-1) which was initially known for its role in the repair of oxidative stress-induced DNA damage, has also been reported to play a mediating role in the inflammatory response. Studies with PARP-1 knockout models have shown that PARP-1 is a co-activator of Nuclear Factor-kappa B (NF-kappaB), although this appears not to require its enzyme activity. In addition, drug-induced inhibition of the enzyme activity of PARP-1 was observed to reduce the production of pro-inflammatory mediators. In this study, the flavonoid compound flavone was demonstrated to significantly inhibit the enzyme activity of PARP-1. Further evaluation of flavone in N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-treated human pulmonary epithelial and vascular endothelial cells revealed that both the decrease in NAD(+) levels, as well as the formation of PAR-polymers was dose-dependently attenuated by flavone. In addition, flavone was found to reduce the lipopolysaccharide (LPS)-induced interleukin (IL)-8 production in pulmonary epithelial cells, which was confirmed by transcription analysis. Furthermore, the transcription Inhibitor kappa B alpha (of IkappaBalpha) was significantly increased by flavone. The results of the present study indicate that the flavonoid flavone could be a potential candidate for application in treatment of chronic inflammatory diseases. PARP-1 inhibition could have beneficial effects in such diseases as Chronic Obstructive Pulmonary Disease (COPD) and diabetes, by preservation of cellular NAD(+) levels and attenuating inflammatory conditions.
Assuntos
Flavonoides/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Inibidores de Poli(ADP-Ribose) Polimerases , Oxirredutases do Álcool/antagonistas & inibidores , Oxirredutases do Álcool/metabolismo , Antioxidantes/química , Antioxidantes/metabolismo , Benzamidas/farmacologia , Linhagem Celular Tumoral , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Óxidos N-Cíclicos/metabolismo , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Compostos Ferrosos/metabolismo , Compostos Ferrosos/farmacologia , Flavonas , Flavonoides/química , Humanos , Peróxido de Hidrogênio/metabolismo , Peróxido de Hidrogênio/farmacologia , Interleucina-8/genética , Interleucina-8/metabolismo , Metilnitronitrosoguanidina/farmacologia , Estrutura Molecular , NAD/metabolismo , NF-kappa B/genética , NF-kappa B/metabolismo , Nucleotidases/farmacologia , Fenantrenos/farmacologia , Poli(ADP-Ribose) Polimerase-1 , Poli(ADP-Ribose) Polimerases/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Detecção de Spin/métodos , Transcrição Gênica/efeitos dos fármacosRESUMO
The activity of the nuclear enzyme poly(ADP-ribose)polymerase-1 (E.C.2.4.2.30), which is highly activated by DNA strand breaks, is associated with the pathophysiology of both acute as well as chronic inflammatory diseases. PARP-1 overactivation and the subsequent extensive turnover of its substrate NAD+ put a large demand on mitochondrial ATP-production. Furthermore, due to its reported role in NF-kappaB and AP-1 mediated production of pro-inflammatory cytokines, PARP-1 is considered an interesting target in the treatment of these diseases. In this study the PARP-1 inhibiting capacity of caffeine and several metabolites as well as other (methyl)xanthines was tested using an ELISA-assay with purified human PARP-1. Caffeine itself showed only weak PARP-1 inhibiting activity, whereas the caffeine metabolites 1,7-dimethylxanthine, 3-methylxanthine and 1-methylxanthine, as well as theobromine and theophylline showed significant PARP-1 inhibiting activity. Further evaluation of these compounds in H2O2-treated A549 lung epithelial and RF24 vascular endothelial cells revealed that the decrease in NAD+-levels as well as the formation of the poly(ADP-ribose)polymer was significantly prevented by the major caffeine metabolite 1,7-dimethylxanthine. Furthermore, H2O2-induced necrosis could be prevented by a high dose of 1,7-dimethylxanthine. Finally, antioxidant effects of the methylxanthines could be ruled out with ESR and measurement of the TEAC. Concluding, caffeine metabolites are inhibitors of PARP-1 and the major caffeine metabolite 1,7-dimethylxanthine has significant PARP-1 inhibiting activity in cultured epithelial and endothelial cells at physiological concentrations. This inhibition could have important implications for nutritional treatment of acute and chronic inflammatory pathologies, like prevention of ischemia-reperfusion injury or vascular complications in diabetes.
Assuntos
Inibidores Enzimáticos/farmacologia , Inibidores de Poli(ADP-Ribose) Polimerases , Teofilina/farmacologia , Xantinas/farmacologia , Difosfato de Adenosina/metabolismo , Trifosfato de Adenosina/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Óxidos N-Cíclicos/farmacologia , Relação Dose-Resposta a Droga , Espectroscopia de Ressonância de Spin Eletrônica/métodos , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Inibidores Enzimáticos/química , Ensaio de Imunoadsorção Enzimática/métodos , Humanos , Peróxido de Hidrogênio/farmacologia , Concentração Inibidora 50 , Estrutura Molecular , NAD/química , NAD/metabolismo , Proteínas Nucleares/antagonistas & inibidores , Proteínas Nucleares/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Poli(ADP-Ribose) Polimerase-1 , Poli(ADP-Ribose) Polimerases/metabolismo , Detecção de Spin/métodos , Relação Estrutura-Atividade , Teofilina/química , Xantinas/químicaRESUMO
Oxidative DNA damage, as occurs during exacerbations in chronic obstructive pulmonary disease (COPD), highly activates the nuclear enzyme poly(ADP-ribose)polymerase-1 (PARP-1). This can lead to cellular depletion of its substrate NAD+, resulting in an energy crisis and ultimately in cell death. Inhibition of PARP-1 results in preservation of the intracellular NAD+ pool, and of NAD+-dependent cellular processes. In this study, PARP-1 activation by hydrogen peroxide decreased intracellular NAD+ levels in human pulmonary epithelial cells, which was found to be prevented in a dose-dependent manner by theophylline, a widely used compound in the treatment of COPD. This enzyme inhibition by theophylline was confirmed in an ELISA using purified human PARP-1 and was found to be competitive by nature. These findings provide new mechanistic insights into the therapeutic effect of theophylline in oxidative stress-induced lung pathologies.