RESUMEN
Intake of antioxidants from the diet has been recognized to have beneficial health effects, but the potential benefit of taking antioxidants such as ß-carotene as supplements is controversial. The aim of the present study was to evaluate the potential protective effects of a physiologically relevant concentration (2 µM) of ß-carotene on the DNA damaging effects of catechol in mouse lymphoma L5178Y cells. Two different exposure protocols were used: simultaneous exposure to ß-carotene and catechol for 3 h; and exposure to catechol for 3 h after 18 h pre-treatment with the vitamin. DNA damage was evaluated using the comet assay (employing one procedure for general damage, and another procedure, which also included oxidative DNA damage). Independent of exposure protocol and procedure for comet assay, ß-carotene did not increase the basal level of DNA damage. However, at the highest concentration of catechol (1 mM), ß-carotene was found to clearly increase the level of catechol-induced DNA damage, especially in the pre-treated cells. Interestingly, an opposite effect was observed at lower concentrations of catechol, but the ß-carotene related reduction of catechol-induced genotoxicity was significant (P < 0.05) only for the procedure including oxidative damage induced by 0.5 mM catechol. Taken together our results indicate that ß- carotene can both reduce and enhance the DNA damaging effects of a genotoxic agent such as catechol. This indicates that it is the level of catechol-induced DNA damage that seems to determine whether ß-carotene should be regarded as a beneficial or detrimental agent when it comes to its use as a dietary supplement.
Asunto(s)
Antioxidantes/farmacología , Daño del ADN/efectos de los fármacos , Estrés Oxidativo , beta Caroteno/farmacología , Animales , Catecoles/farmacología , Suplementos Dietéticos , Humanos , Leucemia L5178 , Ratones , Pruebas de MutagenicidadRESUMEN
AIM OF THE STUDY: Although traditional herbal medicines are widely used in Ethiopia, no information is available on their potential genotoxicity. In the present study, hydroalcoholic extracts of Glinus lotoides, Plumbago zeylanica, Rumex steudelii and Thymus schimperi were evaluated for their DNA damaging effects using the comet assay. MATERIAL AND METHODS: Mouse lymphoma L5178Y cells were exposed to different concentrations of the extracts for 3h with and without metabolic activation (S9-mix) using 4-nitroquinoline-N-oxide and benzo(a)pyrene as positive controls, and vehicles as negative controls. RESULTS: In the absence of S9, all extracts were found to induce significant DNA damage without affecting the cell viability. T. schimperi and R. steudelii were the most potent DNA-damaging extracts, and G. lotoides and P. zeylanica the least potent. The addition of S9 had different effects on the DNA damage induced by the extracts: it lowered the DNA damaging effect of P. zeylanica, did not affect the DNA damaging effect of T. schimperi, and increased the DNA damaging effects of R. steudelii and G. lotoides. CONCLUSION: The findings of the present study suggest that all extracts evaluated have a genotoxic potential in vitro which needs to be substantiated by further studies.
Asunto(s)
Supervivencia Celular/efectos de los fármacos , Daño del ADN , Magnoliopsida , Mutágenos/toxicidad , Extractos Vegetales/toxicidad , 4-Nitroquinolina-1-Óxido/farmacología , Animales , Benzo(a)pireno/farmacología , Línea Celular , Relación Dosis-Respuesta a Droga , Etiopía , Linfoma/patología , Medicinas Tradicionales Africanas , Ratones , Molluginaceae , Pruebas de Mutagenicidad , Extractos Vegetales/química , Plumbaginaceae , Rumex , Thymus (Planta)RESUMEN
The DNA damaging effect of chlorobenzene was investigated in peripheral lymphocytes and bone marrow cells from C57BL/6 female mice using a gel electrophoresis assay for DNA from single cells ('the single cell gel electrophoresis assay') under alkaline conditions. The effect of chlorobenzene was studied both after single and repeated intraperitoneal injections of 750 mg/kg body weight. The cytostatic agent cyclophosphamide (150 mg/kg, i.p.) was used as a reference substance, and vehicle-treated mice as controls. DNA damage was recorded 16 h after the (last) injection, using an automated computerized image analysis system specifically designed for the single cell gel electrophoresis assay. There was evidence of chlorobenzene-induced DNA damage after 3 days of repeated exposure in peripheral lymphocytes, but no indications of such an effect in bone marrow cells. Cyclophosphamide induced significant damage to DNA both in bone marrow cells and lymphocytes, the effect being most pronounced in the latter cells. It is concluded that high-dose exposure to chlorobenzene is associated with genotoxicity to peripheral lymphocytes. However, this solvent is apparently not a major hazard to bone marrow cells, even after repeated high-dose exposure.
Asunto(s)
Clorobencenos/toxicidad , Daño del ADN , Linfocitos T/efectos de los fármacos , Análisis de Varianza , Animales , Médula Ósea/efectos de los fármacos , Células de la Médula Ósea , Recuento de Células/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Clorobencenos/administración & dosificación , Ciclofosfamida/administración & dosificación , Ciclofosfamida/toxicidad , Relación Dosis-Respuesta a Droga , Electroforesis en Gel de Agar , Femenino , Procesamiento de Imagen Asistido por Computador , Inyecciones Intraperitoneales , Ratones , Ratones Endogámicos C57BL , Aceite de Oliva , Aceites de Plantas/administración & dosificación , Estándares de ReferenciaRESUMEN
beta-Cell-rich pancreatic islets from ob/ob mice were taken for measurements of insulin release in response to glucose after culture in RPMI 1640 medium. The stimulatory effect of 20 mmol/l glucose was converted into an inhibition when the medium was supplemented with 400 mumol/l diazoxide. Glucose inhibition of insulin release was observed when the islets had been cultured in the presence of 1 or 20 mmol/l glucose in media either containing or lacking Ca2+. The data provide further evidence for an inhibitory component in the action of glucose on insulin release, suggesting that glucose stimulation of the Ca2+ efflux is essential for the appearance of this inhibition.
Asunto(s)
Diazóxido/farmacología , Glucosa/farmacología , Insulina/metabolismo , Islotes Pancreáticos/metabolismo , Animales , Calcio/metabolismo , Células Cultivadas , Medios de Cultivo , Secreción de Insulina , Ratones , Ratones ObesosRESUMEN
1. Exposure to media deprived of Ca2+ resulted in prompt and transient stimulation of 45Ca efflux from beta cell-rich pancreatic islets microdissected from ob/ob-mice and to some extent also from the isolated neurohypophysis. 2. Particular high efflux rates were reached when the Ca2+-deficient medium contained EGTA, but there was no effect of the chelator on the total amount of radioactivity mobilized from the islets. 3. The removal of extracellular Ca2+ was less effective in promoting the 45Ca efflux in the absence of Na+ and no stimulatory response was seen in the presence of 1 mM-La3+. 4. The 45Ca washout was stimulated whether or not the media used for the loading or subsequent perifusion of the islets were supplemented with 20 mM-D-glucose. However, there was no response to a second exposure to a Ca2+-deficient medium even subsequent to redistribution of intracellular calcium induced by temporary lowering of the temperature. 5. It is suggested that the islet 45Ca released by the removal of extracellular Ca2+ originates from a distinct plasma membrane pool which is exchanged slowly compared to most of the calcium at the beta cell periphery.
Asunto(s)
Calcio/metabolismo , Islotes Pancreáticos/metabolismo , Animales , Transporte Biológico/efectos de los fármacos , Calcio/farmacología , Membrana Celular/metabolismo , Técnicas In Vitro , Ratones , Ratones Obesos , Neurohipófisis/metabolismoRESUMEN
An aqueous extract from the unripe fruits of the tropical plant Momordica charantia was found to be a potent stimulator of insulin release from beta-cell-rich pancreatic islets isolated from obese-hyperglycemic mice. The stimulation of insulin release was partially reversible. It differed from that of D-glucose and other commonly employed insulin secretagogues in not being suppressed by L-epinephrine and in even being potentiated by the removal of Ca2+. This anomalous behaviour was not associated with general effects on the metabolism of the beta-cells as indicated by an unaltered oxidation of D-glucose. Studies of 45Ca fluxes suggest that the insulin-releasing action is the result of perturbations of membrane functions. In support for the idea of direct effects on membrane lipids, the action of the extract was found to mimic that of saponin in inhibiting the Ca2+/H+ exchange mediated by the ionophore A23187 in isolated chromaffin granules and release Ca2+ from preloaded liposomes.