RESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Boldoa purpurascens Cav. (Nyctaginaceae) is a plant species used in traditional medicine in Cuba as a diuretic. AIM OF THE STUDY: The aim of the present investigation was to evaluate the safety profile of a hydroalcoholic extract from leaves of Boldoa purpurascens. MATERIALS AND METHODS: First, an experimental study to assess the oral acute toxicity at a dose of 2000mg/kg body weight of the extract was carried out. Potential genotoxicity of the extract was evaluated using the Ames test and the micronucleus induction assay in mouse bone marrow. In the Ames test a concentration range of 50, 100, 150, 300 and 500µg/plate was tested. In the micronucleus induction assay, doses of 500, 1000 and 2000mg/kg of body weight were tested. For completeness, since the extract contains saponins, the evaluation of the hemolytic activity, ocular and skin irritation were included. RESULTS: No signs or symptoms of toxicity were observed in the oral acute toxicity test (body weight at baseline, seven days and end of the experiment of 236.41±20.07, 256.81±30.44 and 240.02±26.16 respectively for the treated group). The hydroalcoholic extract from the leaves was not mutagenic in the Ames test, and no genotoxicity was observed in the micronucleus assay. A hemolysis test at concentration of 1mg/mL confirmed hemolytic activity, which is not a safety concern since saponins are not absorbed after oral administration. In order to evaluate the percentage of protein denaturation, the ocular irritability index was calculated. The extract was found to be irritating. Finally, skin irritability was evaluated and the irritation index was equal to zero. CONCLUSIONS: Based on the toxicological evaluation of a traditionally used hydroalcoholic extract from the leaves of Boldoa purpurascens we can confirm the safety of its oral use.
Asunto(s)
Nyctaginaceae/toxicidad , Extractos Vegetales/toxicidad , Hojas de la Planta/toxicidad , Administración Oral , Animales , Células de la Médula Ósea/efectos de los fármacos , Células de la Médula Ósea/patología , ADN Bacteriano/efectos de los fármacos , ADN Bacteriano/genética , Dermatitis por Contacto/etiología , Dermatitis por Contacto/patología , Ojo/efectos de los fármacos , Ojo/metabolismo , Proteínas del Ojo/metabolismo , Femenino , Hemólisis/efectos de los fármacos , Masculino , Ratones Endogámicos BALB C , Micronúcleos con Defecto Cromosómico/inducido químicamente , Pruebas de Micronúcleos , Mutación , Nyctaginaceae/química , Fitoterapia , Extractos Vegetales/administración & dosificación , Extractos Vegetales/aislamiento & purificación , Hojas de la Planta/química , Plantas Medicinales , Desnaturalización Proteica , Conejos , Ratas Sprague-Dawley , Ratas Wistar , Medición de Riesgo , Pruebas de Irritación de la Piel , Solventes/química , Factores de Tiempo , Pruebas de Toxicidad AgudaRESUMEN
Cancer cells are particularly vulnerable to treatments impairing redox homeostasis. Reactive oxygen species (ROS) can indeed play an important role in the initiation and progression of cancer, and advanced stage tumors frequently exhibit high basal levels of ROS that stimulate cell proliferation and promote genetic instability. In addition, an inverse correlation between histological grade and antioxidant enzyme activities is frequently observed in human tumors, further supporting the existence of a redox dysregulation in cancer cells. This biochemical property can be exploited by using redox-modulating compounds, which represent an interesting approach to induce cancer cell death. Thus, we have developed a new strategy based on the use of pharmacologic concentrations of ascorbate and redox-active quinones. Ascorbate-driven quinone redox cycling leads to ROS formation and provoke an oxidative stress that preferentially kill cancer cells and spare healthy tissues. Cancer cell death occurs through necrosis and the underlying mechanism implies an energetic impairment (ATP depletion) that is likely due to glycolysis inhibition. Additional mechanisms that participate to cell death include calcium equilibrium impairment and oxidative cleavage of protein chaperone Hsp90. Given the low systemic toxicity of ascorbate and the impairment of crucial survival pathways when associated with redox-active quinones, these combinations could represent an original approach that could be combined to standard cancer therapy.
Asunto(s)
Ácido Ascórbico/farmacología , Neoplasias/tratamiento farmacológico , Estrés Oxidativo/efectos de los fármacos , Quinonas/farmacología , Antineoplásicos , Antioxidantes/uso terapéutico , Ácido Ascórbico/uso terapéutico , Humanos , Neoplasias/metabolismo , Quinonas/uso terapéuticoRESUMEN
Cancer cells show an up-regulation of glycolysis, they readily take up vitamin C, and they appear more susceptible to an oxidative stress than the surrounding normal cells. Here we compare, analyse and discuss these particular hallmarks by performing experiments in murine hepatomas (TLT cells) and freshly isolated mouse hepatocytes. The results show that rates of lactate formation are higher in TLT cells as compared to mouse hepatocytes, but their ATP content represents less than 25% of that in normal cells. The uptake of vitamin C is more important in hepatoma cells as compared to normal hepatocytes. This uptake mainly occurs through GLUT1 transporters. Hepatoma cells have less than 10% of antioxidant enzyme activities as compared to normal hepatocytes. This decrease includes not only the major antioxidant enzymes, namely catalase, superoxide dismutase and glutathione peroxidase, but also the GSH content. Moreover, catalase is almost not expressed in hepatoma cells as shown by western blot analysis. We explored therefore a selective exposure of cancer cells to an oxidative stress induced by pro-oxidant mixtures containing pharmacological doses of vitamin C and a redox active compound such as menadione (vitamin K(3)). Indeed, the combination of vitamin C (which accumulates in hepatoma cells) and a quinone undergoing a redox cycling (vitamin K(3)) leads to an oxidative stress that kills cancer cells in a selective manner. This differential sensitivity between cancer cells and normal cells may have important clinical applications, as it has been observed with other pro-oxidants like Arsenic trioxide, isothiocyanates, Adaphostin.
Asunto(s)
Neoplasias/patología , Estrés Oxidativo , Animales , Línea Celular Tumoral , Humanos , Ratones , Neoplasias/metabolismoRESUMEN
In 2008, we celebrate the 80th anniversary of the discovery of vitamin C. Since then, we know that vitamin C possesses few pharmacological actions although it is still perceived by the public as a "miracle-pill" capable to heal a variety of illnesses. Cancer is one of the most common diseases for which a beneficial role of vitamin C has been claimed. Thus, its dietary use has been proposed in cancer prevention for several years. Apart from this nutritional aspect, an extensive and often confusing literature exists about the use of vitamin C in cancer that has considerably discredited its use. Nevertheless, recent pharmacokinetic data suggest that pharmacologic concentrations of vitamin C can be achieved by intravenous injections. Since these concentrations exhibit anticancer activities in vitro, this raises the controversial question of the re-evaluation of vitamin C in cancer treatment. Therefore, the purpose of this commentary is to make a critical review of our current knowledge of vitamin C, focusing on the rationale that could support its use in cancer therapy.
Asunto(s)
Ácido Ascórbico/uso terapéutico , Neoplasias/tratamiento farmacológico , Ácido Ascórbico/farmacocinética , Ácido Ascórbico/farmacología , Interacciones Farmacológicas , Humanos , Neoplasias/metabolismo , Estrés Oxidativo/efectos de los fármacosRESUMEN
Ageing is an aggravating factor leading to alterations in the biotransformation of drugs, and therefore their therapeutic efficacy and safety. In this review we discuss the influence of ageing on drug metabolizing enzymes in male Wistar rats. We report that drug metabolizing enzymes can be affected by ageing either by post-translational modifications or by transcriptional modifications. The post-translational modifications could be due to an increase of oxidative stress during ageing. Although it is now well established that transcriptional modifications are due to a change in the GH secretion profile in senescent rats, the intracellular mechanisms underlying these modifications are still unclear. In addition to the strong decrease in the activity of the main CYPs of male rats, we discuss the potential consequences on human drug metabolism in the elderly.
Asunto(s)
Sistema Enzimático del Citocromo P-450/metabolismo , Hígado/enzimología , Preparaciones Farmacéuticas/metabolismo , Factores de Edad , Anciano , Animales , Biotransformación , Humanos , Isoenzimas , Hígado/metabolismo , Factores SexualesRESUMEN
The decision of stressed cells to die or to survive is made by integrating signals at different levels through multiple check points. However, initiation and continued progression toward cell death by apoptosis in cancer cells may be blocked by mutation of the tumor suppressor p53 or overexpression of members of the bcl-2 family of proteins. The existence of such mechanisms indicates that cancer cells lose the controls regulating their cell cycle. Therefore, the activation of their programmed cell death appears as a major therapeutic target. Oxidative stress can stimulate growth, trigger apoptosis, or cause necrosis depending upon the dose and the exposure time of the oxidizing agent. A large body of evidence supports the idea that oxidative stress induced by redox cycling of vitamins C and K(3) in association surpasses cancer cellular defense systems and results in cell death. The molecular mechanisms underlying such a process are, however, still unknown. Indeed, several types of cell death may be produced, namely autoschizis, apoptosis and necrosis. Combined vitamin C and K(3) administration in vitro and in vivo produced tumor growth inhibition and increased the life-span of tumor-bearing mice. CK(3)-treatment selectively potentiated tumor chemotherapy, produced sensitization of tumors resistant to some drugs, potentiated cancer radiotherapy and caused inhibition of the development of cancer metastases without inducing toxicity in the host. We propose the association of vitamins C and K(3) as an adjuvant cancer therapy which may be introduced into human cancer therapy without any change in the classical anticancer protocols, and without any supplementary risk for patients.
Asunto(s)
Antioxidantes/uso terapéutico , Ácido Ascórbico/uso terapéutico , Neoplasias/tratamiento farmacológico , Vitamina K 3/uso terapéutico , Animales , Muerte Celular/efectos de los fármacos , Quimioterapia Adyuvante , Sinergismo Farmacológico , Ratones , Neoplasias/patología , Neoplasias/fisiopatología , Estrés Oxidativo/efectos de los fármacos , Células Tumorales CultivadasRESUMEN
The effect of bacterial endotoxin (lipopolysaccharides from Escherichia coli, LPS) on cellular metabolism and drug biotransformation was studied in precision-cut rat liver slices (PCLS). Xenobiotic metabolism by PCLS was assessed by measuring phase I (midazolam hydroxylation) and phase II (paracetamol conjugates) enzyme activities. Nitrites formation was used as an indirect way to assess LPS-mediated activation of nitric oxide synthase (iNOS, type 2). PCLS incubation with various LPS doses results in a dose-dependent formation of nitrites. Such a nitrite formation is decreased by dexamethasone. After incubation of PCLS for 24 h LPS addition did not increase the basal nitrite formation, indicating that cells are not responsive any more. Paracetamol conjugation was unaffected by LPS treatment but midazolam hydroxylation was reduced by more than 50%. Such a loss is not due to cell impairment since neither survival (LDH leakage) nor cellular metabolism (protein synthesis or ATP content) were modified by LPS. Indeed, under defined conditions, namely Williams' medium E and O(2)/CO(2) (95:5), PCLS maintained both ATP and GSH levels and the capacity of hepatocytes to synthesize proteins. In conclusion, the in vitro model of PCLS reproduces the inhibitory effect of LPS on a CYP3A-dependent activity, allowing a mechanistic approach to study cell-cell interactions.