ABSTRACT
Objetivos: establecer un modelo murino de fibrosis pulmonar inducida por bleomicina, investigando el posible papel protector del sistema endocannabinoide (SE) frente a la fibrosis. Métodos: se emplearon ratones salvajes (C5BL/6 y Balb/c) así como la cepa TRPV1-/-. Tras una única dosis intratraqueal de bleomicina, se analizó la respuesta fibrótica mediante un análisis histológico, la determinación de la expresión de marcadores del proceso profibrótico, el estudio de la actividad mieloperoxidasa y del contenido en hidroxiprolina del pulmón, así como el análisis de la expresión génica de VIP, PACAP, IL-1β, IL-6, TNF-α e IL-11, y el estado de activación de las rutas MAPKs (fosfo-JNK, fosfo-ERK) de la ruta de NF-κB (p-IκBα), la ruta de β-catenina y del TGFβ (GSK-3B), la activación de SMAD (pSMAD2) y pSTAT3, a nivel proteico. Resultados: la fibrosis pulmonar inducida por bleomicina en los ratones de la cepa TRPV- /- fue más severa que en la cepa salvaje C5BL/6. El contenido en hidroxiprolina y la actividad mieloperoxidasa fue mayor en los ratones TRPV1-/-. Se detectó un incremento significativo en la expresión génica de citoquinas proinflamatorias (TNF-a, IL-1b, IL11 e IL-6), pero no de VIP o PACAP, en la cepa TRPV1-/-. A nivel proteico, la expresión de pIKBα, pSTAT3, pSMAD2 y pJNK, pero no la de pERK, se vio incrementada en los ratones TRPV1-/-. Conclusiones: el modelo murino de fibrosis pulmonar inducida por bleomicina sigue siendo clave para continuar profundizando los conocimientos acerca de la patogénesis de la FPI. La modulación del SE podría tener un papel protector frente a la fibrosis pulmonar
Objectives: to establish a murine model of bleomycin-induced pulmonary fibrosis, analysing the possible protective role of the Endocannabinoid System (ES) against fibrosis. Methods: wild C5BL/6 and Balb/c mice, as well as the genetically modified strain TRPV1- /- were used. After a single dose of intratracheal bleomycin, the fibrotic response was analysed though histologic studies, the assessment of proinflammatory markers, myeloperoxidase activity, hydroxyproline content, genetic expression of VIP, PACAP, IL-1 β, IL-6, TNF-α and IL-11, as well as MAPK route (phospho-JNK, phospho-ERK), NF-κB (p-IκBα), β-cathenin, TGF-β (GSK-3B), SMAD (p-SMAD2) and pSTAT3, at a protein level. Results: pulmonary fibrosis was more severe in TRPV1-/- mice compared to C5BL/6 mice. A significant increase in proinflammatory markers such as TNF-α, IL-1β, IL11 and IL-6, but not VIP or PACAP, was observed. pIKBα, pSTAT3, pSMAD2 and pJNK, but not pERK, were increased at a protein level in TRPV-/- mice. Conclusions: the murine model of bleomycin-induced lung fibrosis remains a keystone to pioneer current investigation in lung fibrosis. Modulation of the ES might have a protective role
Subject(s)
Animals , Mice , Pulmonary Fibrosis/chemically induced , Pulmonary Fibrosis/veterinary , Bleomycin/therapeutic use , Peroxidase/therapeutic use , Endocannabinoids/therapeutic use , Gene Expression , Models, Animal , Laparotomy/methods , Laparotomy/veterinary , Immunohistochemistry/methods , Blotting, Western/methodsABSTRACT
The intermediate nestin filament is expressed in neural stem cells, neuroectodermal tumors and various adult tissues under situations that reproduce developmental phases, e.g., physiological renewal of certain cell types, tissue regeneration, and healing or revascularization. In the human gastrointestinal tract, nestin has been reported in glial cells and interstitial cells of Cajal. We examined by immunohistochemistry the appearance and distribution of nestin protein in enteric ganglia of rat duodenum. Through the myenteric and submucosal plexuses, a high number of nestin-positive cells were visualized in this specie. The nestin-positive cells were smaller and more numerous than enteric neurons. They were present both within and around ganglia. The results of this study suggest that the rat enteric glial cells (EGCs) are rich in nestin, a protein usually associated with dividing or migrating cells and the dynamic reorganization of nestin filaments during the cell cycle. EGCs could function as enteric stem cells.
Subject(s)
Duodenum/metabolism , Intermediate Filament Proteins/metabolism , Myenteric Plexus/metabolism , Nerve Tissue Proteins/metabolism , Submucous Plexus/metabolism , Animals , Duodenum/chemistry , Duodenum/innervation , Female , Immunohistochemistry , Intermediate Filament Proteins/analysis , Male , Myenteric Plexus/chemistry , Nerve Tissue Proteins/analysis , Nestin , Neuroglia/metabolism , Rats , Rats, Wistar , Submucous Plexus/chemistryABSTRACT
Recently the new term 'telocytes' has been proposed for cells formerly known as interstitial Cajal-like cells. In fact, telocytes are not really Cajal-like cells, they being different from all other interstitial cells by the presence of telopodes, which are cell-body prolongations, very thin, extremely long with a moniliform aspect. The identification of these cells is based on ultrastructural criteria. The presence of telocytes in others organs was previously documented. We reported for the first time, an ultrastructural study of telocytes in the lamina propria of rat duodenum. Our findings show that typical telocytes are present in the rat duodenum. Telocytes are located in the lamina propria, immediately below mucosal crypts. Telopodes frequently establish close spatial relationships with immune cells, blood vessels and nerve endings. On the basis of their distribution and morphology, we suggest that these cells may be involved in immune response and in our opinion, it may be possible that different locations of telocytes could be associated with different roles.
Subject(s)
Duodenum/ultrastructure , Interstitial Cells of Cajal/ultrastructure , Animals , Blood Vessels/ultrastructure , Microscopy, Electron, Transmission , Mucous Membrane/ultrastructure , Rats , Rats, WistarABSTRACT
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