RESUMEN
Biphosphonate (BPN) are widely used in clinics to treat metastatic cancer and osteoporosis thus representing a problem not only for patients but also for workers involved in their preparation and administration. A similar exposure occurred years ago in match-making workers undergoing bone alterations similar to those consequent to BPN exposure. Osteonecrosis of the jaw (ONJ) is a main adverse effect related to BPN administration, which is performed in millions of patients worldwide for osteoporosis and cancer therapy, thus representing an emerging problem in public health. In susceptible patients, BPN induce severe, progressive, and irreversible degeneration of facial bones, resulting in avascular ONJ often triggered by dental surgery. BPN induced ONJ occurs in subjects depending on lifestyle factors of both environmental and endogenous origins. Exogenous risk factors include cigarette smoke, alcohol consumption, bacterial infections, and cyclosporine therapy. Endogenous risk factors include systemic diseases such as diabetes or hypertension and adverse polymorphisms of genes involved in metabolism (CYPs, MTHFR), thrombosis (Factor V, Prothrombin), and detoxification (MDR). Available molecular findings provide evidence that ONJ is related to risk-factors associated with environmental mutagenesis and gene-environment interactions. This issues may be useful to identify susceptible subjects by molecular analyses in order to prevent ONJ occurrence.
Asunto(s)
Osteonecrosis de los Maxilares Asociada a Difosfonatos/etiología , Interacción Gen-Ambiente , Biomarcadores/sangre , Osteonecrosis de los Maxilares Asociada a Difosfonatos/diagnóstico , Colágeno Tipo I/sangre , Ciclosporina/efectos adversos , Daño del ADN , Difosfonatos/efectos adversos , Inhibidores Enzimáticos/efectos adversos , Humanos , Procedimientos Quirúrgicos Orales/efectos adversos , Péptidos/sangre , Polimorfismo de Nucleótido Simple , Radiación Ionizante , Factores de Riesgo , Fumar/efectos adversosRESUMEN
Intracellular RNAses are involved in various functions, including microRNA maturation and turnover. Mutations occurring in genes encoding RNAses cause Aicardi-Goutiéres syndrome (AGS). AGS mutations silence RNAse activity, thus inducing accumulation of endogenous RNAs, mainly consisting of short RNAs and microRNAs. Overload of intracellular RNA triggers Toll like receptor-dependent interferon-alpha production in the brain, which in turn activates neurotoxic lymphocytes and inhibits angiogenesis thus inducing the typical clinical phenotype of AGS. However, these pathogenic mechanisms are attenuated after three years of age by the endogenous production of DNAJP58IPK and Cystatin F, which arrest AGS progression. Because RNAses are involved in microRNA turnover, we evaluated the expression of 957 microRNAs in lymphocytes from AGS patients and control patients. Our results indicate that microRNA overload occurs in AGS patients. This upregulation inhibits microRNA turnover impeding the synthesis of the novel microRNAs required for the differentiation and myelination of the brain during the initial period of postnatal life. These pathogenic mechanisms result in AGS, a neurological syndrome characterized by irritability, mild hyperpyrexia, pyramidal and extrapyramidal signs, and spastic-dystonic tetraplegia. Typical cerebrospinal fluid alterations include lymphocytosis and elevated interferon-alpha levels. Brain imaging demonstrates cerebral calcifications, white matter abnormalities, and progressive cerebral atrophy.Thus, evidence exists that mutations silencing intracellular RNases affect microRNA turnover resulting in the severe clinical consequences in the brain characterizing the clinical feature of AGS.
Asunto(s)
Enfermedades Autoinmunes del Sistema Nervioso/enzimología , Enfermedades Autoinmunes del Sistema Nervioso/genética , Isoenzimas/deficiencia , MicroARNs/metabolismo , Malformaciones del Sistema Nervioso/enzimología , Malformaciones del Sistema Nervioso/genética , Ribonucleasas/deficiencia , Animales , Enfermedades Autoinmunes del Sistema Nervioso/patología , Enfermedades Autoinmunes del Sistema Nervioso/fisiopatología , Niño , ADN/metabolismo , Femenino , Humanos , Isoenzimas/química , Isoenzimas/genética , Masculino , Modelos Moleculares , Malformaciones del Sistema Nervioso/patología , Malformaciones del Sistema Nervioso/fisiopatología , Estructura Terciaria de Proteína , ARN/metabolismo , Ribonucleasas/química , Ribonucleasas/genéticaRESUMEN
In the field of prevention, Internet websites and their related instruments constitute valuable tools for healthcare facilities, and particularly for Local Healthcare Authorities (LHA). As yet, however, their undoubted potential remains largely unexploited. Many LHA websites currently operating in Italy are organized in such a way that they fail to make adequate use of this precious resource. Indeed, communication regarding prevention is all too often limited to the simple reproduction of information and indications in a static and heterogeneous manner, so much so that it resembles a mere "online notice-board". The aim of the present research was to analyze the current situation and the various innovative proposals that have been made, in order to construct a more effective website model that could be used nationwide. To this end, the research was carried out through a two-pronged approach: on the one hand, all 190 LHA websites in Italy were analyzed; on the other, a questionnaire was administered to a sample of habitual users of the most modern and widespread social network, Facebook. Analysis and elaboration of the data gathered led to the creation of the model "@Prevention". This project is intended to introduce an innovative perspective into the field of online communication for healthcare prevention by providing a highly useful tool for the LHA, healthcare workers and, obviously, citizens.
Asunto(s)
Educación en Salud/estadística & datos numéricos , Promoción de la Salud/estadística & datos numéricos , Internet/estadística & datos numéricos , Comercialización de los Servicios de Salud/estadística & datos numéricos , Educación del Paciente como Asunto/estadística & datos numéricos , Humanos , Difusión de la Información/métodos , Relaciones Interpersonales , Italia , Evaluación de Programas y Proyectos de SaludRESUMEN
As the only nourishment and scavenging source for most of the anterior and posterior chamber tissues in the eye, the aqueous humor represents one of the target for glaucoma. The aim of this study is to investigate the yet unexplored relationship between aqueous humor protein content and open-angle glaucoma (POAG) pathogenesis. Aqueous humor was collected from 10 POAG patients (cases) and 14 senile cataract patients (controls), matched for age and gender, undergoing surgery for trabeculectomy and cataract, respectively. Protein samples were cyanine-labeled and hybridized with antibody microarrays. Microarray signals were revealed by laser scanner, quantified, and compared by statistical analyses. Total protein amounts were not significantly different in patients versus controls. Conversely, a proteome cluster significantly modified in patients as compared to controls was identified as highly predictive for disease status. Selected proteins underwent dramatic variation, which was correlated to pathogenetic events characterizing POAG, including oxidative damage, mitochondrial damage, neural degeneration, and apoptosis. The results obtained indicate that proteomic analysis of aqueous humor is a new tool for POAG diagnosis in the case of otherwise uncertain disease recognition. Furthermore, this study allows a better understanding of mechanisms involved in the pathogenesis of POAG, the main cause of irreversible blindness worldwide.
Asunto(s)
Humor Acuoso/química , Proteínas del Ojo/química , Glaucoma de Ángulo Abierto/metabolismo , Proteoma/química , Proteómica/métodos , Anciano , Anciano de 80 o más Años , Análisis de Varianza , Estudios de Casos y Controles , Análisis por Conglomerados , Proteínas del Ojo/clasificación , Proteínas del Ojo/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Análisis de Componente Principal , Análisis por Matrices de Proteínas , Estadísticas no ParamétricasRESUMEN
7,12-Dimethylbenz(a)anthracene (DMBA) is a prototype carcinogen that induces a high yield of mammary tumors in rats after a single feeding. We investigated the induction and chemoprevention of DNA adducts in female Sprague Dawley rats receiving DMBA by gavage according to a variety of treatment schedules. The patterns of 32P-postlabeled DNA adducts in liver and mammary epithelial cells were similar to those produced by the in vitro reaction of metabolically activated DMBA with calf thymus DNA. There was a high and statistically significant correlation between dose of DMBA administered to rats (0, 0.6, 2.4, and 12 mg/kg body weight) and levels of DNA adducts in both types of cells. The regression lines relating DMBA doses to total DNA adduct levels were significantly divergent and crossed at 1.5 mg/kg body weight, indicating that, at lower doses, the formation of DNA adducts is more intense in target mammary cells, whereas at higher doses, DNA adduct levels are more elevated in liver cells, presumably due to the greater metabolic capacity of this organ. When the rats were sacrificed 7 days rather than 2 days after DMBA administration, DNA adduct levels were approximately halved in both liver and mammary cells. The observed patterns can be interpreted based on toxicokinetic factors, local and distant metabolism, removal of DNA adducts by excision repair, and cell proliferation rate. Of three chemopreventive agents given with the diet to rats treated with 12 mg of DMBA, 5,6-benzoflavone (1650 ppm) was the most effective, inhibiting DNA adduct formation in liver and mammary cells by 96.5 and 83.5%, respectively. Feeding of 1,2-dithiole-3-thione (600 ppm) inhibited this biomarker by 68.5 and 50.2%, whereas butyl hydroxyanisole (BHA; 5000 ppm) showed a significant inhibition in the liver (46.5%) but was ineffective in mammary cells (29.0%, not significant). These data correlate nicely with the results of a parallel study in which 5,6-benzoflavone, 1,2-dithiole-3-thione, and BHA inhibited formation of hemoglobin adducts by 80.0, 44.0, and 0%, respectively; the incidence of mammary tumors by 82.4, 47.1, and 5.9%, respectively; and their multiplicity by 92.6, 80.0, and 7.4%, respectively. Therefore, biomarkers of biologically effective dose are highly predictive of the efficacy of chemopreventive agents in the DMBA rat mammary model. The selective inhibition by BHA of DNA adducts in the liver but not in mammary cells is consistent with the finding that this phenolic antioxidant stimulated phase II activities in the liver but not in the mammary gland (L. L. Song et al., manuscript in preparation). In any case, the broad-spectrum inducer 5,6-BF appears to be more effective than the two monofunctional phase II inducers, presumably because an enhanced activation of DMBA to reactive metabolites is coordinated with their blocking, detoxification, and excretion.
Asunto(s)
9,10-Dimetil-1,2-benzantraceno/metabolismo , Anticarcinógenos/farmacología , Carcinógenos/metabolismo , Aductos de ADN/metabolismo , Hígado/efectos de los fármacos , Glándulas Mamarias Animales/efectos de los fármacos , Animales , Hidroxianisol Butilado/farmacología , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Femenino , Glutatión/metabolismo , Hígado/metabolismo , Glándulas Mamarias Animales/metabolismo , Ratas , Ratas Sprague-Dawley , Tionas/farmacología , Tiofenos/farmacologíaRESUMEN
A Phase II chemoprevention trial was carried out in Qidong, Jiangsu Province, People's Republic of China. The recruited subjects, all of whom were positive for serum aflatoxin-albumin adducts, were divided into three treatment arms: placebo; oltipraz ([5-(2-pyrazinyl)-4-methyl-1,2-dithiol-3-thione]) given daily at 125 mg p.o.; and oltipraz given once per week at 500 mg p.o. Besides biomarkers related to aflatoxin B(1) exposure, the genotoxicity of blind-coded urine XAD-2 concentrates was evaluated in 201 subjects on the fifth and seventh week of intervention. Genotoxicity was assessed both in the Ames reversion test in strain YG1024 of Salmonella typhimurium, in the presence of an exogenous metabolic system (S9 mix), with or without beta-glucuronidase, and in a DNA repair test in Escherichia coli. Heating of concentrated urine samples or of cigarette smoke condensates was discovered to result in a significant enhancement of their mutagenicity. It was also found that the mutagenicity of condensates from the most extensively used brands of cigarettes in Qidong was much lower than that of Western cigarette brands. Urine mutagenicity was unrelated to treatment with oltipraz, intervention time, gender, and supplement of S9 mix with beta-glucuronidase. Mutagenicity was significantly but variably higher in cigarette smokers than in nonsmokers, which suggests that the urinary excretion of mutagens in the examined population was not exclusively attributable to smoking. Nevertheless, within smokers (28% of the recruited subjects; 67% of all males), the mutagenic potency was significantly correlated with the self-reported number of cigarettes smoked per day and, even more sharply, with the cotinine concentrations in urines. In conclusion, this study demonstrated the validity of urine mutagenicity assays as a biomarker of tobacco smoke exposure that can be investigated on a relatively large scale in chemoprevention trials and provided evidence that oltipraz treatment had no influence on this parameter in the examined population.
Asunto(s)
Biomarcadores/análisis , Pirazinas/farmacología , Fumar/efectos adversos , Administración Oral , Adulto , Quimioprevención , Escherichia coli/efectos de los fármacos , Escherichia coli/genética , Femenino , Humanos , Masculino , Pruebas de Mutagenicidad , Mutágenos/análisis , Neoplasias/prevención & control , Pirazinas/administración & dosificación , Reproducibilidad de los Resultados , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/genética , Tionas , Tiofenos , OrinaRESUMEN
Reactive oxygen species are involved in the pathogenesis of cancer and other chronic degenerative diseases through a variety of mechanisms, including DNA damage. We investigated by 32p and 33P postlabeling analyses the nucleotidic modifications induced in vitro by treating calf thymus DNA with H2O2 and CuSO4, interacting in a Fenton type reaction. Six different enrichment procedures and three chromatographic systems were comparatively assayed. The chromatographic system using phosphate/urea, which is more suitable for detecting bulky DNA adducts, was rather insensitive. In contrast, the system using acetic acid/ammonium formate revealed high levels of mononucleotidic modifications. In terms of ratio of adduct levels in treated and untreated DNA, the enrichment procedures ranked as follows: nuclease P1 (19.6), no enrichment (18.3), digestion to trinucleotides (17.6), digestion to monophosphate mononucleotides (8.4), digestion to dinucleotides (3.4), and extraction with butanol (<1.0). The system using formic acid/ammonium formate was quite efficient in detecting 8-hydroxy-2'-deoxyguanosine. Labeling with 33p further enhanced the sensitivity of the method. The oxidative damage was so intense to produce a strong DNA fragmentation detectable by agarose gel electrophoresis, and nucleotidic modifications were more intense when DNA fragmentation was greater. The DNA alterations produced by H2O2 alone were significantly lower than those produced following reaction of H2O2 with CuSO4. The thiol N-acetylcysteine (NAC) was quite efficient in inhibiting both nucleotidic modifications and DNA fragmentation produced in vitro by either H2O2 or the .OH generating system. These results support at a molecular level the findings of previous studies showing the ability of NAC to inhibit the genotoxicity of peroxides and of reactive oxygen species generated by electron transfer reactions.
Asunto(s)
Acetilcisteína/farmacología , Aductos de ADN/análisis , Daño del ADN , Depuradores de Radicales Libres/farmacología , Peróxido de Hidrógeno/farmacología , Radical Hidroxilo/farmacología , Especies Reactivas de Oxígeno , Animales , Bovinos , Cromatografía en Capa Delgada , Sulfato de Cobre/farmacología , ADN/efectos de los fármacos , Fragmentación del ADN , Marcaje Isotópico/métodos , Oxidación-Reducción , Radioisótopos de FósforoRESUMEN
Our previous studies showed that nucleotide alterations, evaluated by (32)P postlabeling, are systematically detected in smooth muscle cells of atherosclerotic lesions localized in the aorta of surgical patients. The level of these molecular lesions was correlated with the occurrence of known atherogenic risk factors, among which the number of currently smoked cigarettes, and was significantly enhanced in individuals having a null GSTM1 genotype as compared to individuals carrying the GSTM1 genotype. The present study had the dual objective of evaluating the formation of DNA adducts in the whole thoracic aorta of Sprague-Dawley rats, exposed whole-body to cigarette smoke for 28 consecutive days, and of investigating the effects of chemopreventive agents given orally during the same period. High levels of (32)P postlabeled DNA adducts were formed in the aorta of smoke-exposed rats, with an overall 11 times increase over the total levels observed in sham-exposed rats, and with increases ranging between three and 63 times for seven individual DNA adducts. Supplement of the diet with either 1,2-dithiole-3-thione, phenethyl isothiocyanate or 5,6-benzoflavone had no or poor effects on the smoke-related formation of nucleotide alterations in the aorta. In contrast, oltipraz, given with the diet, N-acetyl-L-cysteine, given with drinking water and, even more potently, their combination exerted remarkable protective effects. The results of this experimental study, together with the previous findings in humans, suggest that DNA alterations may contribute to the atherogenic process, clarify a possible mechanism of cigarette smoke, a well known atherogen, and show the potential protective effects of certain drugs towards these alterations.
Asunto(s)
Aorta Torácica/química , Arteriosclerosis/prevención & control , Aductos de ADN/análisis , Fumar/efectos adversos , Acetilcisteína/uso terapéutico , Animales , Quimioprevención , Ingestión de Alimentos , Masculino , Pirazinas/uso terapéutico , Ratas , Ratas Sprague-Dawley , Tionas , Tiofenos , Aumento de PesoRESUMEN
Experimental data suggest a possible role of DNA damage in aging, mainly related to oxidative lesions. With the objective of evaluating DNA lesions as molecular biomarkers of aging, we measured 8-hydroxy-2'-deoxyguanosine (8-OH-dG) and DNA-protein crosslinks (DPXL) levels in different organs of mice aged 12 and 24 months. 8-OH-dG was detected by 32P postlabelling after removing unmodified dG by trifluoracetic acid, which prevented the artificial formation of 8-OH-dG during 32P labelling procedures. Appreciable 8-OH-dG amounts were detected in 12-month-old mice in liver (1.8 +/- 0.7 8-OH-dG/10(5) normal nucleotides), brain (1.6 +/- 0.5) and heart (2.3 +/- 0.5). In 24-month-old mice these values were higher in all examined organs (liver, 2.7 +/- 0.4; brain, 3.6 +/- 1.1; heart, 6.8 +/- 2.2 8-OH-dG/10(5) normal nucleotides). This accounted for a 1.5-fold increase in liver (not significant), 2.3-fold increase in brain (P < 0.01), and 3.0-fold increase in heart (P < 0.001). A similar trend was observed for DPXL levels, which were the 1.8 +/- 0.3%, 1.2 +/- 0.2%, and 2.2 +/- 0.3% of total DNA in liver, brain, and heart of 12-month-old mice and 1.9 +/- 0.4%, 2.0 +/- 0.4%, and 3.4 +/- 0.5% in 24-month-old mice, with ratios of 1.0, 1.7 (P < 0.01), and 1.5 (P < 0.001), respectively. Highly significant correlations between 8-OH-dG and DPXL levels were recorded in brain (r = 0.619, P < 0.001) and heart (r = 0.800, P < 0.0001), but not in liver (r = 0.201, not significant). These data suggest that brain and heart are more severely affected by the monitored age-related DNA lesions than liver, which can be ascribed to certain characteristics of these postmitotic organs, including the low detoxifying capacities, the high oxygen consumption, and the impossibility to replace damaged cells by mitosis. The strong correlation between 8-OH-dG and DPXL supports a possible contribution of oxidative mechanisms to formation of DPXL in those organs, such as brain and heart, which play a primary role in the aging of the whole organism.
Asunto(s)
Envejecimiento/fisiología , Aductos de ADN/análisis , Desoxiguanosina/análogos & derivados , Proteínas/metabolismo , 8-Hidroxi-2'-Desoxicoguanosina , Factores de Edad , Animales , Biomarcadores , Encéfalo/fisiología , Reactivos de Enlaces Cruzados/análisis , Reactivos de Enlaces Cruzados/metabolismo , Aductos de ADN/química , Aductos de ADN/metabolismo , Daño del ADN/fisiología , Desoxiguanosina/análisis , Desoxiguanosina/metabolismo , Femenino , Corazón/fisiología , Marcaje Isotópico/métodos , Hígado/fisiología , Pulmón/fisiología , Ratones , Ratones Endogámicos , Radioisótopos de Fósforo , Proteínas/química , Ácido Trifluoroacético/químicaRESUMEN
Oxidative damage plays a pathogenic role in various chronic degenerative diseases. Oxidative damage targeting trabecular meshwork (TM) cells as a consequence of mitochondrial damage is a pathogenic mechanism for glaucoma, the most common cause of irreversible blindness worldwide. Consequences of oxidative damage are attenuated by endocellular activities involved in scavenging reactive oxidative species and DNA repair. Selected bacterial genes are highly efficient at protecting cells from oxidative DNA damage. This situation occurs for Escherichia coli formamidopyrimidine DNA glycosylase (FPG), a major DNA glycosylase that repairs oxidatively damaged DNA. Accordingly, this study was aimed at transfecting human TM cells (HTMC) with Fpg in order to increase their resistance to oxidative damage. This study demonstrates that it is feasible to increase resistance of HTMC to endogenous oxidative damage by gene transfection. These findings bear relevance for primary and secondary prevention of degenerative glaucomas and other degenerative diseases where oxidative damage plays a pathogenic role.
Asunto(s)
Daño del ADN , ADN-Formamidopirimidina Glicosilasa/genética , Proteínas de Escherichia coli/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Malla Trabecular/metabolismo , 8-Hidroxi-2'-Desoxicoguanosina , Línea Celular Tumoral , Desoxiguanosina/análogos & derivados , Desoxiguanosina/análisis , Células Endoteliales/metabolismo , Expresión Génica , Terapia Genética , Glaucoma/genética , Glaucoma/prevención & control , Glaucoma/terapia , Humanos , Mitocondrias/metabolismo , Oxidación-Reducción , Estrés Oxidativo/genética , Especies Reactivas de Oxígeno/metabolismo , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/genética , Malla Trabecular/citología , TransfecciónAsunto(s)
Carcinógenos Ambientales/efectos adversos , Predisposición Genética a la Enfermedad/epidemiología , Predisposición Genética a la Enfermedad/genética , ADN/efectos de los fármacos , ADN/genética , Frecuencia de los Genes , Genotipo , Humanos , Biología Molecular , Epidemiología Molecular , Neoplasias/inducido químicamente , Neoplasias/epidemiología , Neoplasias/genética , Neoplasias/prevención & control , Fenotipo , Prevención PrimariaRESUMEN
Reduced glutathione (GSH) plays a critical role as an intracellular defense system providing detoxification of a broad spectrum of reactive species and their excretion as water-soluble conjugates. Conjugation of GSH with electrophiles is catalyzed by GSH S-transferases (GST), which constitute a broad family of phase II isoenzymes. Two of the GST encoding genes, GSTM1 (mu) and GSTT1 (theta), have a null genotype due to their homozygous deletion that results in lack of active protein. Polymorphisms within GSTT1 and especially GSTM1 have often been associated with cancer in various organs as well as with elevated levels of DNA adducts in various cell types. We recently demonstrated that DNA adducts are consistently detectable in smooth muscle cells (SMC) of human abdominal aorta affected by atherosclerotic lesions. Here we provide evidence that levels of adducts to SMC DNA from atherosclerotic lesions are consistently increased in individuals having the null GSTM1 genotype, whereas no association was established with the GSTT1 polymorphism. The influence of GSTM1 deletion was better expressed in never-smokers and ex-smokers than in current smokers. These findings bear relevance to the epidemiology of atherosclerosis and suggest that metabolic polymorphisms may contribute to the interindividual variability in susceptibility not only to carcinogens, but also to DNA binding atherogens.
Asunto(s)
Arteriosclerosis/genética , Aductos de ADN/metabolismo , Desoxiguanosina/análogos & derivados , Glutatión Transferasa/genética , Músculo Liso Vascular/metabolismo , Polimorfismo Genético/genética , 8-Hidroxi-2'-Desoxicoguanosina , Aorta/patología , Arteriosclerosis/metabolismo , Arteriosclerosis/patología , Daño del ADN , Desoxiguanosina/metabolismo , Genotipo , Glutatión Transferasa/metabolismo , Humanos , Isoenzimas/genética , Isoenzimas/metabolismo , Músculo Liso Vascular/patología , Fumar/fisiopatologíaRESUMEN
Multiple drug resistance (MDR) mechanisms are known to limit the effectiveness of some cancer chemotherapies, probably through enhancing P-glycoprotein-mediated drug efflux from mammalian cells. Similar mechanisms appear to act in other organisms, including bacteria, and may affect not only the toxicity but also the mutagenicity of certain chemicals. At least in some experimental situations, MDR can be overcome through concomitant treatment of the cells with various types of inhibitors. Two MDR inhibitors, verapamil, a calcium channel blocker, and trifluoperazine, a calmodulin inhibitor, were assayed for their ability to modulate the potency of nine mutagens with varying mechanisms of action in various Salmonella typhimurium his- strains. Neither verapamil nor trifluoperazine affected the direct mutagenicity of sodium dichromate and 2-methoxy-6-chloro-9[3-(2-chloroethyl)amino-propyl-amino] dihydrochloride (ICR 191) or the S9-mediated mutagenicity of benzo[a]pyrene and 2-amino-3,4-dimethyl-amidazo[4,5-f]quinoline (MeIQ). Both modulators enhanced the direct mutagenicity of doxorubicin. Moreover, trifluoperazine sharply increased the S9-mediated mutagenicity of cyclophosphamide and 2-aminofluorene, while it consistently decreased the mutagenicity of 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2). The contrasting effect towards the aromatic amine 2-aminofluorene and the heterocyclic amine Trp-P-2, representative of important chemical families responsible for the bacterial mutagenicity of cigarette smoke, may explain the observed lack of influence of trifluoperazine on the mutagenicity of a cigarette smoke condensate. These observations extend the known range of chemical types whose mutagenicity can be modulated by inhibitors of MDR and suggest that there may be value in adding MDR inhibitors, especially trifluoperazine, to optimize the detection of mutagenicity by certain types of chemicals in the Salmonella/mammalian microsome mutagenicity test.
Asunto(s)
Bloqueadores de los Canales de Calcio/farmacología , Antagonistas de Dopamina/farmacología , Resistencia a Múltiples Medicamentos/fisiología , Mutagénesis/efectos de los fármacos , Mutágenos/toxicidad , Salmonella typhimurium/efectos de los fármacos , Trifluoperazina/farmacología , Verapamilo/farmacología , Acridinas/toxicidad , Aminacrina/análogos & derivados , Aminacrina/toxicidad , Benzo(a)pireno/antagonistas & inhibidores , Benzo(a)pireno/toxicidad , Cromatos/antagonistas & inhibidores , Cromatos/toxicidad , Ciclofosfamida/antagonistas & inhibidores , Ciclofosfamida/toxicidad , Relación Dosis-Respuesta a Droga , Doxorrubicina/antagonistas & inhibidores , Doxorrubicina/toxicidad , Fluorenos/antagonistas & inhibidores , Fluorenos/toxicidad , Indoles/antagonistas & inhibidores , Indoles/toxicidad , Pruebas de Mutagenicidad , Mutación/efectos de los fármacos , Nicotina/antagonistas & inhibidores , Nicotina/toxicidad , Compuestos de Mostaza Nitrogenada/toxicidad , Quinolinas/antagonistas & inhibidores , Quinolinas/toxicidadRESUMEN
The assessment of pathological effects produced by environmental tobacco smoke in humans is controversial in epidemiological studies. On the other hand, animal models are poorly sensitive to smoke carcinogenicity. We designed an experimental study assessing the tissueselective formation and persistence of DNA adducts in smoke-exposed rats. Sprague-Dawley rats were exposed for 6 h per day, 5 days per week, to environmental smoke resulting from a mixture of sidestream and mainstream smoke generated from Kentucky 2R1 reference cigarettes. The total particulate matter was in the range of 73-93 mg/m(3). DNA adducts were measured by (32)P-post-labelling in rat organs (lung, heart, liver, bladder and testis), tissues (dissected tracheal epithelium) and cells [isolated bronchoalveolar lavage (BAL) cells]. A time-related increase of (32)P-post-labelled DNA modifications was detectable by autoradiography, in the form of massive diagonal radioactive zones and individual spots. Top levels were reached after 4-5 weeks of exposure. The ratio of smoke-induced DNA adducts to the background levels detected in sham-exposed rats was 11.2 in the tracheal epithelium, 10.4 in BAL cells, 7.3 in the heart, 6.3 in the lung, 5.1 in the bladder, 1.9 in the testis and 1. 1 in the liver. Appearance of DNA adducts in the lung was also revealed by synchronous fluorescence spectrophotometry. Smoke-related oxidative damage was demonstrated by a significant enhancement of 8-hydroxy-2'-deoxyguanosine in lung DNA. In parallel, there was a time-related induction of lung microsomal arylhydrocarbon hydroxylase activity, an elevation in cytosolic glutathione S-transferase activity, and a moderate but progressive and significant depletion of reduced glutathione. After discontinuing exposure to environmental cigarette smoke for 1 week, DNA adduct levels significantly dropped in the lung, tracheal epithelium, heart and bladder. The decrease was evident but not statistically significant in BAL cells, and was negligible in the heart. The selective localization and the differential persistence of these promutagenic nucleotide modifications in rat organs, tissues and cells suggest that exposure to environmental cigarette smoke, at least under the high exposure regimens used in experimental studies, may pose a potential risk of developing mutation-related diseases.
Asunto(s)
Aductos de ADN/metabolismo , Pulmón/metabolismo , Nucleótidos/metabolismo , Contaminación por Humo de Tabaco/efectos adversos , 8-Hidroxi-2'-Desoxicoguanosina , Animales , Hidrocarburo de Aril Hidroxilasas/metabolismo , Cromatografía en Capa Delgada , Aductos de ADN/análisis , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , Glutatión Transferasa/metabolismo , Masculino , Especificidad de Órganos , Ratas , Ratas Sprague-Dawley , Factores de TiempoRESUMEN
In spite of the epidemiological evidence supporting a synergism between alcohol consumption and cigarette smoking in the pathogenesis of cancers of the aerodigestive tract, there is a paucity of experimental studies evaluating the effects of these agents under well-controlled conditions and exploring the mechanisms involved. We exposed groups of female BD6 rats, aged 8 months, to ethanol (5% in drinking water for 8 consecutive months) and/or whole-body to mainstream cigarette smoke (1 h/day, 5 days/week for 8 months). DNA was purified from different organs and analyzed for the presence of DNA-protein crosslinks and 32P-postlabeled DNA adducts after butanol enrichment. No significant increase of DNA-protein crosslinks, compared to untreated controls, was induced by any treatment in liver, lung, or heart. 'Spontaneous' nucleotidic modifications were detected by 32P-postlabeling in organs of untreated rats, with the highest levels occurring in the heart. Ingestion of ethanol did not affect DNA adduct levels in any of the organs examined: esophagus, liver, lung, and heart. Exposure to cigarette smoke induced formation of DNA adducts in the lung and heart, but not in the esophagus or liver. The combined ingestion of ethanol resulted in a significant formation of smoke-related DNA adducts in the esophagus and in their further, dramatic increase in the heart. It thus appears that ethanol consumption increases the bioavailability of DNA binding smoke components in the upper digestive tract and favors their systemic distribution. The mechanisms responsible for the interaction between ethanol and smoke and for the selective localization of DNA alterations in different organs are discussed. Formation of DNA adducts in the organs examined may be relevant in the pathogenesis of lung and esophageal cancers as well as in the pathogenesis of other types of chronic degenerative diseases, such as chronic obstructive pulmonary diseases and cardiomyopathies.