RESUMEN
Hyaluronic acids were labeled with a rhenium-tricarbonyl used as single core multimodal probe for imaging and their penetration into human skin biopsies was studied using IR microscopy and fluorescence imaging (labeled SCoMPI). The penetration was shown to be dependent on the molecular weight of the molecule and limited to the upper layer of the skin.
Asunto(s)
Colorantes Fluorescentes/química , Ácido Hialurónico/farmacocinética , Imagen Óptica/métodos , Renio/química , Piel/metabolismo , Humanos , Ácido Hialurónico/análisis , Rayos Infrarrojos , Microscopía/métodos , Microscopía Fluorescente/métodos , Imagen Multimodal/métodos , Absorción Cutánea , Espectroscopía Infrarroja por Transformada de Fourier/métodosRESUMEN
Inorganic complexes are increasingly used for biological and medicinal applications, and the question of the cell penetration and distribution of metallodrugs is key to understanding their biological activity. Oxidative stress is known to be involved in inflammation and in inflammatory bowel diseases for which antioxidative defenses are weakened. We report here the study of the manganese complex Mn1 mimicking superoxide dismutase (SOD), a protein involved in cell protection against oxidative stress, using an approach in inorganic cellular chemistry combining the investigation of Mn1 intracellular speciation using mass spectrometry and of its quantification and distribution using electron paramagnetic resonance and spatially resolved X-ray fluorescence with evaluation of its biological activity. More precisely, we have looked for and found the MS signature of Mn1 in cell lysates and quantified the overall manganese content. Intestinal epithelial cells activated by bacterial lipopolysaccharide were taken as a cellular model of oxidative stress and inflammation. DNBS-induced colitis in mice was used to investigate Mn1 activity in vivo. Mn1 exerts an intracellular antiinflammatory activity, remains at least partially coordinated, with diffuse distribution over the whole cell, and functionally complements mitochondrial MnSOD.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Colitis/tratamiento farmacológico , Modelos Animales de Enfermedad , Enfermedades Inflamatorias del Intestino/tratamiento farmacológico , Compuestos Organometálicos/farmacología , Superóxido Dismutasa/metabolismo , Animales , Antiinflamatorios no Esteroideos/química , Antiinflamatorios no Esteroideos/metabolismo , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Quimiocinas/antagonistas & inhibidores , Quimiocinas/metabolismo , Colitis/inducido químicamente , Colitis/metabolismo , Dinitrofluorobenceno/análogos & derivados , Lipopolisacáridos/antagonistas & inhibidores , Lipopolisacáridos/farmacología , Masculino , Ratones , Ratones Endogámicos C57BL , Estructura Molecular , Compuestos Organometálicos/química , Compuestos Organometálicos/metabolismo , Estrés Oxidativo/efectos de los fármacos , Superóxido Dismutasa/químicaRESUMEN
A combinatorial approach using a one-bead-one-compound method and a screening based on a SOD-activity assay was set up for the discovery of an efficient peptidyl copper complex. The complex exhibited good stability constants, suitable redox potentials and excellent intrinsic activity. This complex was further assayed in cells for its antioxidant properties and showed beneficial effects when cells were subjected to oxidative stress.
Asunto(s)
Materiales Biocompatibles/metabolismo , Cobre/química , Péptidos/química , Secuencia de Aminoácidos , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Colon/citología , Colon/efectos de los fármacos , Colon/metabolismo , Cobre/metabolismo , Células HT29 , Humanos , Interleucina-8/metabolismo , Lipopolisacáridos/toxicidad , Estrés Oxidativo/efectos de los fármacos , Péptidos/metabolismo , Superóxido Dismutasa/metabolismoRESUMEN
A conjugate of a Mn-based superoxide dismutase mimic with a Re-based multimodal probe 1[combining low line] was studied in a cellular model of oxidative stress. Its speciation was investigated using Re and Mn X-fluorescence. Interestingly, 1[combining low line] shows a distribution different from its unconjugated analogue but a similar concentration in mitochondria and a similar bioactivity.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Complejos de Coordinación/farmacología , Renio/farmacología , Superóxido Dismutasa/metabolismo , Antiinflamatorios no Esteroideos/química , Antiinflamatorios no Esteroideos/metabolismo , Complejos de Coordinación/química , Complejos de Coordinación/metabolismo , Humanos , Estructura Molecular , Estrés Oxidativo/efectos de los fármacos , Renio/química , Renio/metabolismoRESUMEN
A superoxide dismutase mimic (Mn1) was functionalized with three positively charged-peptides: RRRRRRRRR (Mn1-R9), RRWWWRRWRR (Mn1-RW9) or Fx-r-Fx-K (Mn1-MPP). Characterization of the physico-chemical properties of the complexes show that they share similar binding affinity for Mn2+, apparent reduction potential and intrinsic superoxide dismutase activity. However, their accumulation in cells is different (Mn1-R9 < Mn1-MPP < Mn1-RW9 < Mn1), as well as their subcellular distribution. In addition, the three functionalized-complexes display a better anti-inflammatory activity than Mn1 when assayed at 10 µM. This improvement is due to a combination of an anti-inflammatory effect of the peptidyl moiety itself, and of the SOD mimic for Mn1-RW9 and Mn1-MPP. In contrast, the enhanced anti-inflammatory activity of Mn1-R9 is solely due to the SOD mimic.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Péptidos de Penetración Celular/farmacología , Superóxido Dismutasa/metabolismo , Antiinflamatorios no Esteroideos/química , Antiinflamatorios no Esteroideos/metabolismo , Péptidos de Penetración Celular/química , Péptidos de Penetración Celular/metabolismo , Células HT29 , Humanos , Lipopolisacáridos/antagonistas & inhibidores , Lipopolisacáridos/farmacología , Estructura Molecular , Superóxido Dismutasa/química , TermodinámicaRESUMEN
Macrophages are key players of immunity that display different functions according to their activation states. In a regenerative context, pro-inflammatory macrophages (Ly6Cpos) are involved in the mounting of the inflammatory response whereas anti-inflammatory macrophages (Ly6Cneg) dampen the inflammation and promote tissue repair. Reactive oxygen species (ROS) production is a hallmark of tissue injury and of subsequent inflammation as described in a bacterial challenge context. However, whether macrophages produce ROS following a sterile tissue injury is uncertain. In this study, we used complementary in vitro, ex vivo and in vivo experiments in mouse to show that macrophages do not release ROS following a sterile injury in skeletal muscle. Furthermore, expression profiles of genes involved in the response to oxidative stress in Ly6Cpos and Ly6Cneg macrophage subsets did not indicate any antioxidant response in this context. Finally, in vivo, pharmacological antioxidant supplementation with N-Acetyl-cysteine (NAC) following skeletal muscle injury did not alter macrophage phenotype during skeletal muscle regeneration. Overall, these results indicate that following a sterile injury, macrophage-derived ROS release is not involved in the regulation of the inflammatory response in the regenerating skeletal muscle.
Asunto(s)
Antioxidantes/metabolismo , Macrófagos/metabolismo , Músculo Esquelético/lesiones , Músculo Esquelético/fisiología , Regeneración/fisiología , Animales , Inflamación/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Especies Reactivas de Oxígeno/metabolismo , Superóxido Dismutasa/metabolismo , Superóxidos/metabolismoRESUMEN
SOD-mimics are small complexes that reproduce the activity of superoxide dismutases, natural proteins that catalytically dismutate the superoxide anion. Activated macrophages, which produce ROS and RNS fluxes, constitute a relevant model to challenge antioxidant activity in a cellular context and were used to test a Mn-complex which was shown to efficiently alter the flow of O(2)(-), ONOO(-) and H(2)O(2).