RESUMO
Oxidative damage has been implicated in the pathogenesis of Parkinson disease (PD) but the literature data are confusing. Using products of lipid and DNA oxidation measured by accurate methods, we assessed the extent of oxidative damage in PD patients. The levels of plasma F(2)-isoprostanes (F(2)-IsoPs), hydroxyeicosatetraenoic acid products (HETEs), cholesterol oxidation products, neuroprostanes (F(4)-NPs), phospholipase A(2) (PLA(2)) and platelet activating factor-acetylhydrolase (PAF-AH) activities, urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG), and serum high-sensitivity C-reactive protein were compared in 61 PD patients and 61 age-matched controls. The levels of plasma F(2)-IsoPs, HETEs, 7beta-and 27-hydroxycholesterol, 7-ketocholesterol, F(4)-NPs, and urinary 8-OHdG were elevated, whereas the levels of plasma PLA(2) and PAF-AH activities were lower, in PD patients compared to controls (p< 0.05). The levels of plasma F(2)-IsoPs, HETEs, and urinary 8-OHdG were higher in the early stages of PD (p trend< 0.05). There was a significant negative correlation between the cumulative intake of levodopa and urinary 8-OHdG (r= -0.305, p= 0.023) and plasma total HETEs (r= -0.285, p= 0.043). Oxidative damage markers are systemically elevated in PD, which may give clues about the relation of oxidative damage to the onset and progression of PD.
Assuntos
Biomarcadores/metabolismo , Estresse Oxidativo , Doença de Parkinson/sangue , Doença de Parkinson/diagnóstico , Idoso , Proteína C-Reativa/química , Estudos de Casos e Controles , Colesterol/química , DNA/química , Progressão da Doença , F2-Isoprostanos/química , Feminino , Humanos , Ácidos Hidroxieicosatetraenoicos/química , Levodopa/farmacologia , Lipídeos/química , Masculino , Pessoa de Meia-Idade , Oxigênio/química , Doença de Parkinson/patologiaRESUMO
Oxidative stress and oxidative modification of biomolecules are involved in several physiological and pathophysiological processes. We have previously reported that Notopterygium forbesii Boiss (NF), a traditional Chinese medicine, and its active constituents, including phenethyl ferulate (PF), bergaptol, and isoimperatorin, induced oxidative stress with increased levels of reactive species and heme oxygenase-1 in human fetal hepatocytes. The current study determined the effects of NF and PF on the inflammatory effects of lipopolysaccharide (LPS). Exposure of RAW 264.7 macrophages to LPS increased the expression of inducible nitric oxide synthase and cyclooxygenase 2 and stimulated the formation of reactive nitrogen species. In a coculture system, the LPS-activated macrophages also induced expression of cell adhesion molecules (including E-selectin, intercellular cell adhesion molecule 1, and vascular cell adhesion molecule 1) in human umbilical vein endothelial cells (HUVEC) and vascular smooth muscle cells (VSMC). Preincubation of macrophages with NF or PF attenuated the effects of LPS on macrophages as well as their effects on HUVEC and VSMC. These inhibitory effects of NF and PF were decreased in the presence of N-acetyl-l-cysteine (NAC). At the same time, NAC also reduced NF- or PF-induced increases in reactive oxygen species (ROS) and Hsp32 protein levels and the formation of protein carbonyls in the macrophages. These results suggest that NF- or PF-induced ROS generation and oxidative modifications of intracellular proteins may be responsible for the inhibitory actions of NF and PF on LPS-induced inflammatory responses. These data add to the growing literature that ROS may sometimes be anti-inflammatory.
Assuntos
Anti-Inflamatórios/farmacologia , Apiaceae/química , Ácidos Cumáricos/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Células Cultivadas , Ácidos Cumáricos/química , Ácidos Cumáricos/isolamento & purificação , Cisteína/metabolismo , Dinoprostona/farmacologia , Heme Oxigenase-1 , Hepatócitos/efeitos dos fármacos , Humanos , Macrófagos/metabolismo , Óxido Nítrico/biossíntese , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Óxido Nítrico Sintase Tipo II/farmacologia , Estresse Oxidativo/fisiologia , Extratos Vegetais/química , Extratos Vegetais/farmacologiaRESUMO
This study investigated the effect of a single dose of tomato sauce on healthy male volunteers in a randomized crossover study. Healthy male subjects (n = 10) were enrolled. Placebo (rice and olive oil) or tomato (tomato sauce, rice and olive oil) meals were provided to the volunteers. Blood and urine samples were taken before consumption of meal (0 h) and 2, 4, 6, 24 and 48 h after meal. Consumption of tomato sauce increased plasma lycopene level by 5-22%, with a maximum level at 24 h (p<0.01) after the meal. Levels of plasma F(2)-isoprostanes, hydroxyeicosatetraenoic acid products, allantoin and urinary 8-hydroxy-2'-deoxyguanosine did not change after either meal, but urinary F(2)-isoprostanes (p<0.05) significantly decreased at 48 h compared to 0 h after the tomato sauce meal. This study showed that a single dose of tomato sauce meal had only a limited antioxidant effect in vivo.
Assuntos
Antioxidantes/administração & dosagem , Biomarcadores/análise , Carotenoides/sangue , Solanum lycopersicum , Adulto , Alantoína/sangue , Cromatografia Líquida de Alta Pressão , Estudos Cross-Over , Desoxiadenosinas/urina , Dieta , F2-Isoprostanos/sangue , F2-Isoprostanos/urina , Humanos , Ácidos Hidroxieicosatetraenoicos/sangue , Licopeno , MasculinoRESUMO
Notopterygium forbesii Boiss (NF) has been used as a traditional Chinese medicine for the treatment of common cold and rheumatism. However, there has been limited research on the biological properties of NF, and the mechanisms of action remain unknown. Here, we aimed to study the mechanism of NF-induced heme oxygenase-1 (HO-1) in human fetal hepatocytes (HFHs) and to identify the constituents responsible. Exposure of HFHs to NF causes oxidative stress with the accumulation of reactive species, which in turn leads to the phosphorylation of p38 MAPK and nuclear accumulation of Nrf2 transcription factor, and eventually increased levels of HO-1 mRNA and protein. The increases in reactive species and HO-1 protein are inhibited by agonists of glucocorticoid receptors (GR), such as RU28362, prednisolone, and dexamethasone, as well as by N-acetyl-L-cysteine and SB203580 (a p38 inhibitor), suggesting a role of GR in NF-induced increases in reactive species and HO-1. Assay-guided fractionation of NF led to three active compounds, phenethyl ferulate, bergaptol, and isoimperatorin, that were found to increase oxidative stress and HO-1 protein levels in HFHs. The induction of HO-1 protein in response to moderate oxidative stress may explain some of the beneficial pharmacological effects of NF.
Assuntos
Medicamentos de Ervas Chinesas/química , Heme Oxigenase-1/biossíntese , Hepatócitos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Plantas Medicinais/química , Agonistas Adrenérgicos beta/farmacologia , Androstanóis/farmacologia , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Fracionamento Químico , Indução Enzimática , Expressão Gênica/efeitos dos fármacos , Inativação Gênica , Heme Oxigenase-1/antagonistas & inibidores , Heme Oxigenase-1/genética , Hepatócitos/enzimologia , Humanos , Fígado/citologia , Fígado/embriologia , Fator 2 Relacionado a NF-E2/genética , Estresse Oxidativo/efeitos dos fármacos , Fosforilação , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Soy sauce is a traditional fermented seasoning in Asian countries, that has high antioxidant activity in vitro and some antioxidant activity in vivo. We attempted to identify the major antioxidants present, using the 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay as a guide. 3-Hydroxy-2-methyl-4H-pyran-4-one (maltol) was one of several active compounds found in an ethyl acetate extract of dark soy sauce (DSS) and was present at millimolar concentrations in DSS. However, most of the antioxidant activity was present in colored fractions, two of which (CP1 and CP2) were obtained by gel filtration chromatography. Their structural characteristics based on nuclear magnetic resonance (NMR) and electrospray-ionization time-of-flight mass spectrometry (ESI-TOF-MS) analysis suggest that carbohydrate-containing pigments such as melanoidins are the major contributors to the high antioxidant capacity of DSS.
Assuntos
Antioxidantes/análise , Análise de Alimentos/métodos , Alimentos de Soja/análise , Acetatos/química , Antioxidantes/química , Cromatografia em Gel , Cromatografia Líquida de Alta Pressão , Sequestradores de Radicais Livres , Espectroscopia de Ressonância Magnética , Peptídeos/química , Polímeros/química , Pironas/química , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
Attack on DNA by some reactive nitrogen species results in deamination of adenine and guanine, leading to the formation of hypoxanthine and xanthine, respectively. Published levels of these products in cellular DNA have varied widely. Although these two deamination products are often measured by GC-MS analysis, the procedure of acid hydrolysis to release DNA bases for derivatization poses a risk of artifactual deamination of the DNA. In this study, we demonstrated the artifactual formation of these two deamination products during acid hydrolysis and hence developed a method for detecting and measuring 2'-deoxyinosine, the nucleoside of hypoxanthine. Our assay for 2'-deoxyinosine employs nuclease P1 and alkaline phosphatase to achieve release of the nucleosides from DNA, followed by HPLC prepurification with subsequent GC-MS analysis of the nucleosides. This assay detected an increase in the levels of 2'-deoxyinosine in DNA when commercial salmon testis DNA was treated with nitrous acid. We also used it to measure levels in various rat tissues of both normal and endotoxin-treated rats, but could not find increased 2'-deoxyinosine formation in tissues even though *NO production was substantially increased.
Assuntos
Artefatos , DNA/metabolismo , Aminação , Animais , Bovinos , Cromatografia Líquida de Alta Pressão , Modelos Animais de Doenças , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Hidrólise , Hipoxantina/metabolismo , Células Jurkat , Lipopolissacarídeos/farmacologia , Óxido Nítrico/biossíntese , Estresse Oxidativo , Ratos , Sepse/metabolismo , Xantina/metabolismoRESUMO
Dark soy sauce (DSS) is a powerful antioxidant in vitro. We investigated whether this effect could occur in vivo and improve vascular function. Healthy human subjects were given DSS or placebo meals in a randomized, crossover study. Blood and urine were sampled before and 1, 2, 3, and 4h after the meal for F(2)-isoprostanes (total, free, and esterified) and 8OHdG measurements. Blood pressure, vascular augmentation index (AIx), and heart rate (HR) were also measured. Plasma total F(2)-isoprostanes significantly decreased 3h after placebo and the decrease was greater for DSS. Plasma free and esterified F(2)-isoprostanes were also significantly decreased after DSS. Both placebo and DSS meals increased urinary F(2)-isoprostanes at 1h but not thereafter, and lowered urinary 8OHdG levels, DBP and AIx, and increased HR. We conclude that DSS decreases lipid peroxidation in vivo. However, oxidative damage biomarkers changed after the placebo meal, a phenomenon to consider when designing interventional studies.