RESUMO
Natural antioxidants and vitamins have potential to protect biological systems from peroxidative damage induced by peroxyl radicals, α-tocopherol (Vitamin E, lipid soluble) and ascorbic acid (vitamin C, water soluble), well known natural antioxidant molecules. In the present study we described the synthesis and biological evaluation of hybrid of these two natural antioxidants with each other via ammonium di-ethylether linker, Toc-As in gene delivery. Two control cationic lipids N14-As and Toc-NOH are designed in such a way that one is with ascorbic acid moiety and no tocopherol moiety; another is with tocopherol moiety and no ascorbic acid moiety respectively. All the three cationic lipids can form self-assembled aggregates. The antioxidant efficiencies of the three lipids were compared with free ascorbic acid. The cationic lipids (Toc-As, N14-As and Toc-NOH) were formulated individually with a well-known fusogenic co-lipid DOPE and characterization studies such as DNA binding, heparin displacement, size, charge, circular dichroism were performed. The biological characterization studies such as cell viability assay and in vitro transfection studies were carried out with the above formulations in HepG2, Neuro-2a, CHO andHEK-293T cell lines. The three formulations showed their transfection efficiencies with highest in Toc-As, moderate inN14-As and least in Toc-NOH. Interestingly, the transfection efficiency observed with the antioxidant based conjugated lipid Toc-As is found to be approximately two and half fold higher than the commercially available lipofectamine 2000 at 4:1 charge ratio in Hep G2 cell lines. In the other cell lines studied the efficiency of Toc-As is found to be either higher or similarly active compared to lipofectamine 2000. The physicochemical characterization results show that Toc-As lipid is showing maximum antioxidant potency, strong binding with pDNA, least size and optimal zeta potential. It is also found to be least toxic in all the cell lines studied especially in Neuro-2a cell lines when compared to other two lipids. In summary, the designed antioxidant lipid can be exploited as a delivering system for treating ROS related diseases such as malignancy, brain stroke, etc.
Assuntos
Ácido Ascórbico/farmacologia , DNA/química , Sequestradores de Radicais Livres/farmacologia , Lipossomos/farmacologia , Tensoativos/farmacologia , alfa-Tocoferol/farmacologia , Animais , Ácido Ascórbico/síntese química , Ácido Ascórbico/química , Ácido Ascórbico/toxicidade , Células CHO , Linhagem Celular Tumoral , Cricetulus , DNA/genética , Sequestradores de Radicais Livres/síntese química , Sequestradores de Radicais Livres/química , Sequestradores de Radicais Livres/toxicidade , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Humanos , Lipossomos/síntese química , Lipossomos/química , Lipossomos/toxicidade , Camundongos , Tensoativos/síntese química , Tensoativos/química , Tensoativos/toxicidade , Transfecção/métodos , alfa-Tocoferol/síntese química , alfa-Tocoferol/química , alfa-Tocoferol/toxicidadeRESUMO
The unique properties such as high optical clarity, stability and enhanced bioavailability of nanoemulsion make them useful for food, cosmetic and pharmaceutical industries. In this work, sodium stearoyl lactate and Tween 80 surfactants were collectively used to fabricate alpha tocopherol based oil in water nanoemulsion using high energy ultrasonication method. The spherical nature of pure and drug loaded nanoemulsion has been confirmed with transmission electron microscopy (TEM). The influence of pH, dilution, surfactant concentration and ionic strength on average particle size of pure and nutraceutical (benzylisothiocyanate and curcumin) encapsulated emulsion was examined. The prepared emulsion exhibited good stability up to 90days in salt solution (50-200mM) and different pH conditions. The cumulative release % of benzylisothiocyanate and curcumin was found to be 50.29% in 36h and 89.15% in 150h respectively. The antioxidant activity of pure, benzylisothiocyanate, curcumin and cocktail (benzylisothiocyanate and curcumin) nanoemulsion was calculated with 2,2-diphenyl-1-picrylhydrazyl radical. The IC50 value of different antioxidant showed that benzylisothiocyanate nanoemulsion acted as better antioxidant as compared to pure and curcumin encapsulated nanoemulsion. Also the cell viability of pure nanoemulsion was found to be 24% on hep G2 cell. The effect of UV light irradiation on curcumin and benzylisothiocyanate stability was carried out in different solvent conditions (water/ethanol and nanoemulsion). The degradation of curcumin by the impact of UV light was successfully controlled by trapping in NEm.
Assuntos
Fenômenos Químicos , Portadores de Fármacos/química , Nanoestruturas/química , Óleos/química , Estearatos/química , Água/química , alfa-Tocoferol/química , Compostos de Bifenilo/química , Sobrevivência Celular/efeitos dos fármacos , Curcumina/química , Portadores de Fármacos/toxicidade , Liberação Controlada de Fármacos , Emulsões , Células Hep G2 , Humanos , Concentração de Íons de Hidrogênio , Isotiocianatos/química , Concentração Osmolar , Tamanho da Partícula , Picratos/química , Polissorbatos/química , Sonicação , Tensoativos/química , alfa-Tocoferol/toxicidadeRESUMO
Oxidative stress affects bone turnover. Preventative effects of antioxidants such as vitamin E on reduced bone mineral density and fractures associated with aging, osteoporosis, and smoking have been examined in animals and humans. The effects of vitamin E (α-tocopherol; αT) on bone health have yielded conflicting and inconclusive results from animal studies. In this study, to determine the bone effects of αT, we investigated the in vivo effects of αT on the bone mineral density, bone mass, bone microstructure, bone resorption, and osteogenesis through peripheral quantitative computed tomography (pQCT) measurements, micro-computed tomography (micro-CT) analyses, and bone histomorphometry of lumbar vertebrae and femurs in normal female Wistar rats fed diets containing αT in different quantities (0, 30, 120, or 600 mg/kg diet) for 8 weeks. To validate our hypotheses regarding bone changes, we examined ovariectomized rats as an osteoporosis model and control sham-operated rats in parallel. As expected, ovariectomized rats had reduced bone mineral density in lumbar vertebrae and the distal metaphyses of their femurs, reduced bone mass and deteriorated microstructure of cancellous bones in the vertebral body and distal femur metaphyses, and reduced bone mass due to resorption-dominant enhanced bone turnover in secondary cancellous bones in these sites. In comparison, αT administered to normal rats, even at the highest dose, did not induce reduced bone mineral density of lumbar vertebrae and femurs or a reduced bone mass or fragile microstructure of cancellous bones of the vertebral body and distal femur metaphyses. Instead, αT-fed rats showed a tendency for an osteogenesis-dominant bone mass increase in secondary cancellous bones in the vertebral body, in which active bone remodeling occurs. Thus, αT consumption may have beneficial effects on bone health.
Assuntos
Antioxidantes/toxicidade , Doenças Ósseas Metabólicas/induzido quimicamente , alfa-Tocoferol/toxicidade , Animais , Antioxidantes/administração & dosagem , Peso Corporal/efeitos dos fármacos , Densidade Óssea/efeitos dos fármacos , Remodelação Óssea/efeitos dos fármacos , Reabsorção Óssea/induzido quimicamente , Suplementos Nutricionais , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Comportamento Alimentar/efeitos dos fármacos , Feminino , Fêmur/química , Fêmur/diagnóstico por imagem , Fêmur/ultraestrutura , Humanos , Imageamento Tridimensional , Vértebras Lombares/química , Vértebras Lombares/diagnóstico por imagem , Vértebras Lombares/ultraestrutura , Osteogênese/efeitos dos fármacos , Osteoporose Pós-Menopausa , Ovariectomia , Estresse Oxidativo , Ratos , Ratos Wistar , Microtomografia por Raio-X , alfa-Tocoferol/administração & dosagemRESUMO
The vitamin E family includes both tocopherols and tocotrienols, where α-tocopherol (αTOC) is the most bioavailable form. Clinical trials testing the therapeutic efficacy of high-dose αTOC against stroke have largely failed or reported negative outcomes when a "more is better" approach to supplementation (>400 IU/d) was used. This work addresses mechanisms by which supraphysiologic αTOC may contribute to stroke-induced brain injury. Ischemic stroke injury and the neuroinflammatory response were studied in tocopherol transfer protein-deficient mice maintained on a diet containing αTOC vitamin E at the equivalent human dose of 1680 IU/d. Ischemic stroke-induced brain injury was exacerbated in the presence of supraphysiologic brain αTOC levels. At 48 h after stroke, S100B and RAGE expression was increased in stroke-affected cortex of mice with elevated brain αTOC levels. Such increases were concomitant with aggravated microglial activation and neuroinflammatory signaling. A poststroke increase in markers of oxidative injury and neurodegeneration in the presence of elevated brain αTOC establish that at supraphysiologic levels, αTOC potentiates neuroinflammatory responses to acute ischemic stroke. Exacerbation of microglial activation by excessive αTOC likely depends on its unique cell signaling regulatory properties independent of antioxidant function. Against the background of clinical failure for high-dose αTOC, outcomes of this work identify risk for exacerbating stroke-induced brain injury as a result of supplementing diet with excessive levels of αTOC.
Assuntos
Antioxidantes/toxicidade , Lesões Encefálicas/induzido quimicamente , Inflamação/induzido quimicamente , Isquemia/complicações , Microglia/patologia , Acidente Vascular Cerebral/complicações , alfa-Tocoferol/toxicidade , Animais , Biomarcadores/metabolismo , Lesões Encefálicas/metabolismo , Lesões Encefálicas/patologia , Humanos , Técnicas Imunoenzimáticas , Inflamação/metabolismo , Inflamação/patologia , Isquemia/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microglia/efeitos dos fármacos , Microglia/metabolismo , Acidente Vascular Cerebral/patologia , Superóxidos/metabolismoRESUMO
PURPOSE: Dry eye (DE) includes a group of diseases related to tear film disorders. Current trends for DE therapy focus on providing lipid components to replace the damaged lipid layer. Formulations that contain aqueous and mucin-like compounds may have additional therapeutic benefits for DE patients. The aim of this work was to design and evaluate novel formulations having the potential to become topical treatment for DE. METHODS: Unpreserved liposomal formulations composed of phosphatidylcholine (PC), cholesterol, and α-tocopherol (vit E) were prepared by the thin-film hydration technique. Formulations were characterized in terms of liposome size, pH, surface tension, osmolarity, and viscosity. In vitro tolerance assays were performed on macrophage, human corneal, and conjunctival cell lines at short- and long-term exposures. In vivo ocular tolerance was studied after instillation of the formulation. RESULTS: The mean liposome size was less than 1 µm and surface tension < 30 mN/m for all formulations. The final liposomal formulation (PC-cholesterol-vit E in a ratio of 8:1:0.8) had physiological values of pH (6.45 ± 0.09), osmolarity (289.43 ± 3.28 mOsm), and viscosity (1.82 ± 0.02 mPa · s). Cell viability was greater than 80% in the corneal and conjunctival cells. This formulation was well tolerated by experimental animals. CONCLUSIONS: The unpreserved liposomal formulation has suitable properties to be administered by a topical ophthalmic route. The liposome-based artificial tear had good in vitro and in vivo tolerance responses. This formulation, composed of a combination of liposomes and bioadhesive polymers, may be used successfully as a tear film substitute in DE therapy.
Assuntos
Síndromes do Olho Seco/tratamento farmacológico , Lipossomos/administração & dosagem , Administração Tópica , Animais , Antioxidantes/administração & dosagem , Antioxidantes/toxicidade , Células Cultivadas , Colesterol/administração & dosagem , Colesterol/toxicidade , Túnica Conjuntiva/citologia , Córnea/citologia , Modelos Animais de Doenças , Humanos , Lipossomos/efeitos adversos , Lipossomos/toxicidade , Macrófagos/efeitos dos fármacos , Camundongos , Fosfatidilcolinas/administração & dosagem , Fosfatidilcolinas/toxicidade , Coelhos , Viscosidade , alfa-Tocoferol/administração & dosagem , alfa-Tocoferol/toxicidadeRESUMO
The safety and pharmacokinetic profile of liposomal formulations containing combinations of the antioxidants α-tocopherol, γ-tocopherol or N-acetylcysteine in beagle dogs was examined. Each group consisted of beagle dogs of both genders with a control group receiving empty dipalmitoylphosphatidylcholine (DPPC) liposomes (330 mg/kg DPPC, EL), and test groups receiving liposomes prepared from DPPC lipids with (i) N-acetylcysteine (NAC) (60 mg/kg NAC [L-NAC]); (ii) NAC and α-tocopherol (αT) (60 mg/kg NAC and 25 mg/kg α-tocopherol [L-αT-NAC]) and (iii) NAC and γ-tocopherol (60 mg/kg NAC and 25 mg/kg γ-tocopherol (γT) [L-γT-NAC]). The dogs in the control group (EL) and three test groups exhibited no signs of toxicity during the dosing period or day 15 post treatment. Weight gain, feed consumption and clinical pathology findings (hematology, coagulation, clinical chemistry, urinalysis) were unremarkable in all dogs and in all groups. Results from the pharmacokinetic study revealed that the inclusion of tocopherols in the liposomal formulation significantly increased the area under the curve (AUC) and ß-half life for NAC; the tocopherols had greater impact on the clearance of NAC, where reductions of central compartment clearance (CL) ranged from 56% to 60% and reductions of tissue clearance (CL2) ranged from 73% to 77%. In conclusion, there was no treatment-related toxicity in dogs at the maximum feasible dose level by a single bolus intravenous administration while the addition of tocopherols to the liposomal formulation prolonged the circulation of NAC in plasma largely due to a decreased clearance of NAC.
Assuntos
Acetilcisteína , Antioxidantes , alfa-Tocoferol , gama-Tocoferol , 1,2-Dipalmitoilfosfatidilcolina/química , Acetilcisteína/administração & dosagem , Acetilcisteína/farmacocinética , Acetilcisteína/toxicidade , Animais , Antioxidantes/administração & dosagem , Antioxidantes/farmacocinética , Antioxidantes/toxicidade , Química Farmacêutica , Cães , Relação Dose-Resposta a Droga , Estabilidade de Medicamentos , Feminino , Injeções Intravenosas , Lipossomos , Masculino , Taxa de Depuração Metabólica , Testes de Toxicidade , alfa-Tocoferol/administração & dosagem , alfa-Tocoferol/farmacocinética , alfa-Tocoferol/toxicidade , gama-Tocoferol/administração & dosagem , gama-Tocoferol/farmacocinética , gama-Tocoferol/toxicidadeRESUMO
Liposomes have been used for the delivery of antioxidants to different tissues and organs for the treatment of oxidative stress-induced injuries. In this study, the acute toxicity of a single dose of intravenously (i.v.) administered liposomal antioxidant formulation, containing N-acetylcysteine (NAC) with or without α-tocopherol (α-T) or γ-tocopherol (γ-T), in rats was examined. Each group consisted of 5 male and 5 female Sprague-Dawley rats, with a control group receiving empty dipalmitoylphosphatidylcholine (DPPC) liposomes (660 mg/kg) and test groups receiving DPPC liposomes (660 mg/kg) entrapped with 1) NAC (200 mg/kg), 2) NAC (200 mg/kg) and α-T (83.3 mg/kg), and 3) NAC (200 mg/kg) and γ-T (71.4 mg/kg). These dose levels were determined from the dose-range-finding study and were considered to be the maximum feasible dose (MFD) levels, based on the volume of 10 mL/kg and physical properties and viscosity of the test articles that could be safely administered to rats by an i.v. injection. Two weeks after treatment (day 15), rats in the control group and three test groups exhibited no clinical signs of toxicity during the dosing period or during the 14-day post-treatment period. Weight gain and food consumption in all animals was appropriate for the age and sex of animals. Clinical pathology findings (e.g., hematology, coagulation, clinical chemistry, and urinalysis) were unremarkable in all rats and in all groups. In conclusion, the results of this study showed no treatment-related toxicity in rats at the MFD level by a single bolus i.v. administration.
Assuntos
Acetilcisteína/administração & dosagem , Acetilcisteína/toxicidade , Antioxidantes/química , Lipossomos/química , Testes de Toxicidade Aguda , alfa-Tocoferol/toxicidade , gama-Tocoferol/toxicidade , Animais , Antioxidantes/administração & dosagem , Química Farmacêutica , Feminino , Injeções Intravenosas , Lipossomos/administração & dosagem , Masculino , Ratos , Ratos Sprague-Dawley , alfa-Tocoferol/administração & dosagem , gama-Tocoferol/administração & dosagemRESUMO
α-Tocopheryl succinate (α-TOS) inhibits oxidative phosphorylation at the level of mitochondrial complex I and II, thus promoting cancer cell death through mitochondrial reactive oxygen species (ROS) generation. Redox imbalance activates NF-E2 p45-related factor 2 (Nrf2), a transcription factor involved in cell protection and detoxification responses. Here we examined the involvement of heme oxygenase-1 (HO-1) in the regulation of nuclear factor κB (NF-κB) signaling by short exposure to α-TOS in prostate cancer cells. A short-term (4 h) exposure to α-TOS causes a significant reduction in cell viability (76%±9%) and a moderate rise in ROS production (113%±8%). α-TOS alters glutathione (GSH) homeostasis by inducing a biphasic effect, i.e., an early (1 h) decrease in intracellular GSH content (56%±20%) followed by a threefold rise at 4 h. α-TOS increases nuclear translocation and electrophile-responsive/antioxidant-responsive elements binding activity of Nrf2, resulting in up-regulation of downstream genes cystine-glutamic acid exchange transporter and HO-1, while decreasing NF-κB nuclear translocation. This effect is suppressed by the pharmacological inhibition of HO-1 and mimicked by the end-products of HO activity, i.e., bilirubin and carbon monoxide. Results suggest a little understood mechanism for α-TOS-induced inhibition of NF-κB nuclear translocation due to HO-1 up-regulation.
Assuntos
Heme Oxigenase-1/metabolismo , Proteínas de Membrana/metabolismo , NF-kappa B/metabolismo , alfa-Tocoferol/farmacologia , Animais , Antineoplásicos/farmacologia , Antineoplásicos/toxicidade , Antioxidantes/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Glutationa/metabolismo , Heme Oxigenase-1/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fator 2 Relacionado a NF-E2/metabolismo , NF-kappa B/antagonistas & inibidores , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Transporte Proteico/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , alfa-Tocoferol/toxicidadeRESUMO
Coal mining and incineration of solid residues of health services (SRHS) generate several contaminants that are delivered into the environment, such as heavy metals and dioxins. These xenobiotics can lead to oxidative stress overgeneration in organisms and cause different kinds of pathologies, including cancer. In the present study the concentrations of heavy metals such as lead, copper, iron, manganese and zinc in the urine, as well as several enzymatic and non-enzymatic biomarkers of oxidative stress in the blood (contents of lipoperoxidation = TBARS, protein carbonyls = PC, protein thiols = PT, α-tocopherol = AT, reduced glutathione = GSH, and the activities of glutathione S-transferase = GST, glutathione reductase = GR, glutathione peroxidase = GPx, catalase = CAT and superoxide dismutase = SOD), in the blood of six different groups (n = 20 each) of subjects exposed to airborne contamination related to coal mining as well as incineration of solid residues of health services (SRHS) after vitamin E (800 mg/day) and vitamin C (500 mg/day) supplementation during 6 months, which were compared to the situation before the antioxidant intervention (Ávila et al., Ecotoxicology 18:1150-1157, 2009; Possamai et al., Ecotoxicology 18:1158-1164, 2009). Except for the decreased manganese contents, heavy metal concentrations were elevated in all groups exposed to both sources of airborne contamination when compared to controls. TBARS and PC concentrations, which were elevated before the antioxidant intervention decreased after the antioxidant supplementation. Similarly, the contents of PC, AT and GSH, which were decreased before the antioxidant intervention, reached values near those found in controls, GPx activity was reestablished in underground miners, and SOD, CAT and GST activities were reestablished in all groups. The results showed that the oxidative stress condition detected previously to the antioxidant supplementation in both directly and indirectly subjects exposed to the airborne contamination from coal dusts and SRHS incineration, was attenuated after the antioxidant intervention.
Assuntos
Poluentes Ocupacionais do Ar/toxicidade , Antioxidantes/uso terapêutico , Minas de Carvão , Suplementos Nutricionais , Estresse Oxidativo , Ácido Ascórbico/administração & dosagem , Ácido Ascórbico/uso terapêutico , Biomarcadores/sangue , Estudos de Casos e Controles , Exposição Ambiental , Glutationa/sangue , Glutationa/toxicidade , Glutationa Redutase/sangue , Glutationa Redutase/toxicidade , Humanos , Incineração , Peroxidação de Lipídeos , Metais Pesados/toxicidade , Metais Pesados/urina , Carbonilação Proteica , Superóxido Dismutase/sangue , Superóxido Dismutase/toxicidade , Substâncias Reativas com Ácido Tiobarbitúrico/toxicidade , Vitamina E/administração & dosagem , Vitamina E/uso terapêutico , alfa-Tocoferol/sangue , alfa-Tocoferol/toxicidadeRESUMO
Mitocans are drugs selectively killing cancer cells by destabilizing mitochondria and many induce apoptosis via generation of reactive oxygen species (ROS). However, the molecular events by which ROS production leads to apoptosis has not been clearly defined. In this study with the mitocan alpha-tocopheryl succinate (alpha-TOS) the role of the Bcl-2 family proteins in the mechanism of malignant cell apoptosis has been determined. Exposure of several different cancer cell lines to alpha-TOS increased expression of the Noxa protein, but none of the other proteins of the Bcl-2 family, an event that was independent of the cellular p53 status. alpha-TOS caused a profound conformational change in the pro-apoptotic protein, Bak, involving oligomerization in all cell types, and this also applied to the Bax protein, but only in non-small cell lung cancer cells. Immunoprecipitation studies indicated that alpha-TOS activates the two BH1-3 proteins, Bak or Bax, to form high molecular weight complexes in the mitochondria. RNAi knockdown revealed that Noxa and Bak are required for alpha-TOS-induced apoptosis, and the role of Bak was confirmed using Bak- and/or Bax-deficient cells. We conclude that the major events induced by alpha-TOS in cancer cells downstream of ROS production leading to mitochondrial apoptosis involve the Noxa-Bak axis. It is proposed that this represents a common mechanism for mitochondrial destabilization activated by a variety of mitocans that induce accumulation of ROS in the early phases of apoptosis.
Assuntos
Antineoplásicos/toxicidade , Apoptose , Mitocôndrias/efeitos dos fármacos , alfa-Tocoferol/toxicidade , Proteína Killer-Antagonista Homóloga a bcl-2/metabolismo , Antineoplásicos/química , Humanos , Células Jurkat , Proteínas de Transporte da Membrana Mitocondrial/metabolismo , Membranas Mitocondriais/metabolismo , Poro de Transição de Permeabilidade Mitocondrial , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Proteína Supressora de Tumor p53/genética , alfa-Tocoferol/química , Proteína Killer-Antagonista Homóloga a bcl-2/química , Proteína Killer-Antagonista Homóloga a bcl-2/genéticaRESUMO
Excess hepatic iron generates reactive oxygen species that result in oxidative stress and oxidative damage to the liver. Vitamins have hitherto been considered to be a possible remedy. The aim of this study was to determine if high doses of delta-alpha-tocopherol supplementation in iron overload would ameliorate the oxidative stress. Four groups of 20 male Wistar albino rats were studied: group 1 (control) was fed normal diet, group 2 (Fe) 0.75% Ferrocene iron, group 3 (FV gp) 0.75% Ferrocene/delta-alpha-tocopherol (10x RDA), group 4 (V gp) normal diet/delta-alpha-tocopherol. After 12 months, serum iron, reduced glutathione, catalase, vitamin C, Oxygen Radical Absorbance Capacity, lipid peroxidation, 8-hydroxy-2'-deoxyguanosine (8-OHdG), aspartate transaminase (AST), and alanine transaminase (ALT) were measured. Vitamin C levels were: F gp = 5.04 +/- 0.09; FV gp = 5.85 +/- 0.13 (micromol/l) (p < 0.05). 8-hydroxy-2'-deoxyguanosine levels were: F gp = 143.6 +/- 6.4; FV gp = 179.2 +/- 18.2 (ng/ml) (p < 0.05). Oxidative liver damage, as determined by serum AST and ALT levels, was not attenuated by alpha-tocopherol. A positive correlation existed between vitamin C and 8-OHdG, suggesting possible delta-alpha-tocopherol toxicity.
Assuntos
Antioxidantes/toxicidade , Ácido Ascórbico/metabolismo , Desoxiguanosina/análogos & derivados , Sobrecarga de Ferro/induzido quimicamente , Fígado/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Vitaminas/toxicidade , alfa-Tocoferol/toxicidade , 8-Hidroxi-2'-Desoxiguanosina , Administração Oral , Alanina Transaminase/sangue , Animais , Antioxidantes/administração & dosagem , Ácido Ascórbico/sangue , Aspartato Aminotransferases/sangue , Biomarcadores/sangue , Catalase/sangue , Desoxiguanosina/metabolismo , Modelos Animais de Doenças , Compostos Ferrosos , Glutationa/sangue , Ferro/sangue , Sobrecarga de Ferro/metabolismo , Sobrecarga de Ferro/patologia , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Masculino , Metalocenos , Ratos , Ratos Wistar , Fatores de Tempo , Regulação para Cima , Vitaminas/administração & dosagem , alfa-Tocoferol/administração & dosagemRESUMO
Rats were fed diets containing 0%, 1%, 3%, or 5% mixed tocopheryl phosphates for 90 days. No abnormal clinical signs related to treatment appeared. Some statistically significant changes in hematology and clinical chemistry parameters appeared, but the majority were not dose dependent, occurred in only one sex or group, and/or remained within the historical control range for this strain of rat. A statistically significant apparent reduction in blood protein was observed in animals treated with the tocopheryl phosphates, but further investigation showed that the test substance interfered with the protein assay. Repeat analysis using a method unaffected by plasma test substance levels showed no difference in plasma proteins among all groups. Gross necropsy revealed no abnormalities; reduced relative heart and epididymal weights were observed, but were not dose dependent and were considered incidental. Histopathological changes occurred only in the mesenteric lymph node and small intestine. Foreign material in a crystal-like form appeared in macrophages in both organs, and increased in a dose-related fashion. In the lymph node, sinus histiocytosis increased with dose, but the severity was similar between the control and low-dose groups. Foreign-body granulomatous inflammation, associated with Maltese cross birefringence of the crystals was seen in the mid- and high-dose animals, but not the low-dose group. Similarly, the small intestine showed increasing amounts of foreign material and inflammation in the mid- and high-dose but not in the 1% diet. The 1% diet (equivalent to 587 and 643 mg mixed tocopheryl phosphates/kg body weight/day for male and female rats, respectively) was considered the no observed adverse effect level.
Assuntos
Testes de Toxicidade Crônica/métodos , alfa-Tocoferol/análogos & derivados , Administração Oral , Animais , Peso Corporal/efeitos dos fármacos , Química Clínica/métodos , Cristalização , Dieta , Relação Dose-Resposta a Droga , Ingestão de Líquidos/efeitos dos fármacos , Ingestão de Alimentos/efeitos dos fármacos , Epididimo/efeitos dos fármacos , Epididimo/patologia , Feminino , Coração/efeitos dos fármacos , Hematologia/métodos , Fígado/efeitos dos fármacos , Fígado/patologia , Linfonodos/efeitos dos fármacos , Linfonodos/patologia , Masculino , Mesentério/efeitos dos fármacos , Mesentério/patologia , Miocárdio/patologia , Nível de Efeito Adverso não Observado , Tamanho do Órgão/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Baço/efeitos dos fármacos , Baço/patologia , Urinálise/métodos , alfa-Tocoferol/administração & dosagem , alfa-Tocoferol/química , alfa-Tocoferol/toxicidadeRESUMO
Alpha-tocopheryl succinate (TOS) is a well-known potent and selective apoptotic agent. This apoptotic activity has been ascribed to its detergent-like property which is also shared by the structurally related compound, alpha-tocopheryl phosphate (TOP). TOP meets the structural requirements that have been described for the apoptotic activity of TO esters, i.e. the combination of three structural, one functional, one signalling and one hydrophobic domain. In this study, we have investigated the effect of TOP on the osteosarcoma cell line MG-63 using TOS as a reference compound. As compared with TOS, TOP showed a higher proliferative and apoptosis inducing activity on the MG-63 cancer cell line. The cytotoxic effect of TOP and TOS seems to be due to the effect of the intact compounds, since only a minor conversion into alpha-tocopheryl (TO) could be detected. EPR experiments showed that TOS and TOP reduced membrane fluidity, whereas TO had no effect. In addition, induction of erythrocyte hemolysis by TOP depended on the pH. These results suggest that the detergent-like activity of these compounds might be involved in their biological effect. Due to the potent biological activities, TOP might be clinically useful.
Assuntos
Apoptose , Inibidores do Crescimento/toxicidade , alfa-Tocoferol/análogos & derivados , Linhagem Celular Tumoral , Inibidores do Crescimento/química , Hemólise/efeitos dos fármacos , Humanos , Fluidez de Membrana/efeitos dos fármacos , Tocoferóis , Vitamina E/análogos & derivados , Vitamina E/química , Vitamina E/toxicidade , alfa-Tocoferol/química , alfa-Tocoferol/toxicidadeRESUMO
We have shown that phenolic antioxidant tocopherols are oxidized to nonarylating alpha-tocopheryl quinone (alpha-TQ) and arylating gamma- and delta-TQ electrophiles. The arylating quinones stimulate apoptosis and are highly cytotoxic in mammalian cells. Some xenobiotic phenolic antioxidants are mutagens, and it has been suggested that their arylating quinone metabolites are the active agents in mutagenesis related to carcinogenesis. We found that neither alpha- nor gamma-TQ was directly genotoxic in supercoiled-to-nicked circular DNA conversions, but these agents interacted with the cytomegalovirus reporter-driven plasmid and enhanced luciferase transfection, with gamma-TQ > alpha-TQ. The Ames test, using gamma-TQ and a number of Salmonella strains, showed no evidence of bacterial mutagenesis. gamma-TQ was highly cytotoxic and alpha-TQ slightly cytotoxic in eukaryocyte AS52 cells. A guanosine phosphoribosyltransferase gene assay showed that gamma-TQ was highly mutagenic and alpha-TQ slightly mutagenic in AS52 cells. A review of the literature identified associations where a decrease in dietary gamma-tocopherol (gamma-T) diminishes and an increase in dietary gamma-T and its quinone enhances carcinogenicity. Humans and other omnivores selectively accumulate alpha-tocopherol, even though gamma-T is their principal dietary tocopherol. We suggest that this selectivity confers an evolutionary advantage by limiting tissue gamma-T, a putative precursor of the mutagen gamma-TQ.
Assuntos
Antioxidantes/toxicidade , Mutagênese/fisiologia , Vitamina E/análogos & derivados , Vitamina E/toxicidade , alfa-Tocoferol/toxicidade , Análise Mutacional de DNA , Células Eucarióticas , Humanos , Luciferases/farmacologia , Testes de Mutagenicidade , Plasmídeos/genética , Fatores de Tempo , Transfecção , Células Tumorais CultivadasRESUMO
The effects of sodium nitrite (NaNO2), in combination with one of three antioxidants, tert-butylhydroquinone (TBHQ), alpha-tocopherol (alpha-Toc) and propyl gallate (PG), on N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) stomach carcinogenesis were investigated in F344 rats. Groups of 15 male rats were treated with an intragastric dose of 150 mg/kg body weight of MNNG, and starting 1 week later, were treated with 0.5% TBHQ, 1% alpha-Toc, 1% PG or basal diet with or without 0.2% NaNO2 in their drinking water until they were killed at the end of week 36. Macroscopically, in MNNG-treated animals, combined administration of alpha-Toc or PG with NaNO2 significantly increased the areas and numbers of forestomach nodules as compared with the respective antioxidant alone values. Microscopically, in MNNG-treated animals, treatment with TBHQ significantly increased the incidence and multiplicity of forestomach papillomas as compared with basal diet alone value. Combined administration of alpha-Toc with NaNO2 significantly raised the multiplicity of forestomach papillomas, with a tendency to elevation in the incidence as compared with the group given alpha-Toc alone. Incidences of forestomach moderate and/or severe hyperplasias were significantly higher in the TBHQ or PG plus NaNO2 groups than in the single compound groups. In rats without MNNG treatment, combined treatment of antioxidants with NaNO2 significantly increased the incidences of mild or moderate hyperplasia. In the glandular stomach, although the incidence of atypical hyperplasia showed a non-significant tendency for decrease with TBHQ treatment, additional administration of NaNO2 caused significant increase. These results indicate that co-administration of NaNO2 with alpha-Toc, TBHQ or PG and particularly the first, promotes forestomach carcinogenesis. Concurrent alpha-Toc, TBHQ or PG treatment with NaNO2 is likely to induce forestomach tumors in the long term.