RESUMEN
During recent years, significant development has been achieved in carbon nanotube conjugated with polymer system for drug delivery system (DDS). In the present study, we have prepared functionalized single walled carbon nanotube conjugated with chitooligosaccharide (f-SWNT-COS) as a Drug Delivery System. In addition, drug Gliotoxin (GTX) and targeting molecules (Lysozyme, p53 and Folic acid) have been incorporated into f-SWNT-COS. f-SWNTs-COS-GTX-p53, f-SWNTs-COS-GTX-lysozyme, f-SWNTs-COS-GTX-FA have been physiochemically characterized for DDS. FT-IR, SEM and TEM analysis confirmed the formation of chemical interaction and polymer coating. FT-IR result clearly confirmed the interaction between f-SWNT and COS. The effective drug release was monitored against cervical cancer (HeLa) cells and Breast Cancer (MCF-7) cells and it was found that all the three drug delivery systems showed significant cytotoxicity. f-SWNTs-COS-GTX-p53 delivery vehicle and its effective cytotoxicity on HeLa cells was further checked with fluorescent activated cell sorter analysis. Our results suggest that the f-SWNTs-COS-GTX-p53 is the most effective delivery vehicle with a controlled release and enhanced cytotoxicity rendered through apoptosis in human cervical cancer (HeLa) cells. These systems can further be used for the delivery of other commercially available anti cancer drugs as well.
Asunto(s)
Antineoplásicos/administración & dosificación , Portadores de Fármacos/química , Gliotoxina/administración & dosificación , Nanotubos de Carbono/química , Neoplasias/tratamiento farmacológico , Polímeros/química , Antineoplásicos/farmacocinética , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Quitina/química , Portadores de Fármacos/síntesis química , Portadores de Fármacos/farmacocinética , Evaluación Preclínica de Medicamentos , Gliotoxina/farmacocinética , Células HeLa , Humanos , Células MCF-7 , Neoplasias/metabolismo , Oligosacáridos/químicaRESUMEN
Gliotoxin has been shown to promote a reversal of liver fibrosis in an animal model of the disease although its mechanism of action in the liver is poorly defined. The effects of gliotoxin on activated hepatic stellate cells (HSCs) and hepatocytes have therefore been examined. Addition of gliotoxin (1.5 microM) to culture-activated HSCs resulted in its rapid accumulation, resulting in increased levels of glutathione and apoptosis without any evidence of oxidative stress. In contrast, although hepatocytes also rapidly sequestered gliotoxin, cell death only occurred at high (50-microM) concentrations of gliotoxin and by necrosis. At high concentrations, gliotoxin was metabolized by hepatocytes to a reduced (dithiol) metabolite and glutathione was rapidly oxidized. Fluorescent dye loading experiments showed that gliotoxin caused oxidative stress in hepatocytes. Antioxidants--but not thiol redox active compounds--inhibited both oxidative stress and necrosis in hepatocytes. In contrast, HSC apoptosis was not affected by antioxidants but was potently abrogated by thiol redox active compounds. The adenine nucleotide transporter (ANT) is implicated in mitochondrial-dependent apoptosis. HSCs expressed predominantly nonliver ANT isoform 1, and gliotoxin treatment resulted in a thiol redox-dependent alteration in ANT mobility in HSC extracts, but not hepatocyte extracts. In conclusion, these data suggest that gliotoxin stimulates the apoptosis of HSCs through a specific thiol redox-dependent interaction with the ANT. Further understanding of this mechanism of cell death will aid in finding therapeutics that specifically stimulate HSC apoptosis in the liver, a promising approach to antifibrotic therapy.
Asunto(s)
Gliotoxina/farmacología , Cirrosis Hepática/patología , Hígado/efectos de los fármacos , Hígado/patología , Animales , Antioxidantes/farmacología , Células Cultivadas , Relación Dosis-Respuesta a Droga , Gliotoxina/administración & dosificación , Gliotoxina/farmacocinética , Gliotoxina/toxicidad , Glutatión/metabolismo , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , Hepatocitos/patología , Hígado/metabolismo , Masculino , Necrosis , Ratas , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno/metabolismo , Sustancias Reductoras/farmacología , Compuestos de Sulfhidrilo/antagonistas & inhibidoresRESUMEN
A strain of Aspergillus fumigatus has been isolated from sediments of a mussel bed. When cultured in hyper saline conditions (with sea-water), it produces a cytotoxic and immunosuppressive toxin, gliotoxin, which is excreted in an exudate. In order to know if this toxin could represent a risk for shellfish consumers, an experiment of bioaccumulation of gliotoxin in mussel has been carried out. After 6 days of contamination, toxin was accumulated in the meat of the mussels, at a level up to 2.9 microg/mg of extract weight, with a mode of contamination different to the classical digestive process described for a majority of marine toxins, but similar to the contamination mode of domoic acid.
Asunto(s)
Aspergillus fumigatus/aislamiento & purificación , Bivalvos/metabolismo , Exposición a Riesgos Ambientales/análisis , Gliotoxina/farmacocinética , Toxinas Marinas/farmacocinética , Contaminantes del Agua/análisis , Animales , Bivalvos/química , Monitoreo del Ambiente , Contaminación de Alimentos , Sedimentos Geológicos/microbiología , Gliotoxina/análisis , Gliotoxina/metabolismo , Toxinas Marinas/análisis , Alimentos Marinos , Agua de MarRESUMEN
The fungal metabolite gliotoxin is characterized by an internal disulfide bridge and can exist in either disulfide or dithiol forms. Gliotoxin and other members of the epipolythiodioxopiperazine class of toxins have immunosuppressive properties and have been implicated in human and animal mycotoxicoses. The bridged disulfide moiety is thought to be generally essential for biological activity. Here we show that only the natural (oxidized) form of gliotoxin is actively concentrated in a cell line in a glutathione-dependent manner. Intracellular levels of the toxin can be up to 1500-fold greater than the applied concentration, and toxin in the cells exists almost exclusively in the reduced form. A simple model of toxin entry followed by reduction to the cell-impermeant dithiol explains active uptake, cell density dependence of EC50 values and predicts a value for the maximum concentration of toxin at limiting cell density in agreement with the experiment. Oxidation of the intracellular toxin results in rapid efflux from the cell that also occurs when glutathione levels fall following induction of apoptotic cell death by the toxin. This mechanism allows for minimal production of the toxin while enabling maximal intracellular concentration and thus maximal efficacy of killing in a competitor organism initially present at low cell density. The toxin effluxes from the apoptotic cell exclusively in the oxidized form and can further enter and kill neighboring cells, thus acting in a pseudocatalytic way.