RESUMO
The misuse of anabolic androgenic steroid associated or not with physical workouts disrupts gastrointestinal (GI) function homeostasis. Our goal was to investigate the effects of nandrolone decanoate (ND) and moderate swimming on the GI transit of solid meals, GI motor contractility, and intestinal histology in rats. Male Wistar rats were allocated to four groups that received intramuscular injections of ND (5.0 mg/kg) or vehicle (60.0 µL) and were submitted or not to swimming sessions (60 min, 5% body weight overload) for 4 weeks. Gastric emptying, intestinal transit, in vitro GI contractility, intestinal morphometry, and duodenal mucosal mast cells were evaluated in all experimental groups. ND treatment accelerated gastric emptying, slowed small intestine transit time, enhanced gastric carbachol-mediated reactivity, decreased crypt depth and villus height, reduced mucosal thickness, and increased the circular and longitudinal muscle layer thickness of the duodenum in sedentary rats. Moderate exercise accelerated intestinal transit time and reduced submucosa thickness. In vehicle-treated animals, a strong negative correlation was found between intestinal transit and mucosal mast cells, which was reversed by ND treatment. Combining ND treatment and swimming accelerated gastric emptying, increased duodenal cholinergic reactivity, inhibited the sodium nitroprusside relaxing response, increased the number of duodenal mast cells, decreased villus height, and increased the thickness of all muscle layers. ND changed the morphological and functional properties of the GI tract over time, with intense dysmotility, especially in sedentary animals, but moderate exercise seemed to have played a compensatory role in these harmful effects in the gut.
Assuntos
Anabolizantes , Duodeno , Motilidade Gastrointestinal , Decanoato de Nandrolona , Nandrolona , Condicionamento Físico Animal , Ratos Wistar , Animais , Masculino , Decanoato de Nandrolona/farmacologia , Duodeno/efeitos dos fármacos , Motilidade Gastrointestinal/efeitos dos fármacos , Anabolizantes/farmacologia , Nandrolona/farmacologia , Nandrolona/análogos & derivados , Mastócitos/efeitos dos fármacos , Ratos , Natação , Esvaziamento Gástrico/efeitos dos fármacos , Mucosa Intestinal/efeitos dos fármacos , Trânsito Gastrointestinal/efeitos dos fármacosRESUMO
Mast cells have long been recognized for their involvement in allergic pathology through the immunoglobulin E (IgE)-mediated degranulation mechanism. However, there is growing evidence of other "non-canonical" degranulation mechanisms activated by certain pathogen recognition receptors. Mast cells release several mediators, including histamine, cytokines, chemokines, prostaglandins, and leukotrienes, to initiate and enhance inflammation. The chemical nature of activating stimuli influences receptors, triggering mechanisms for the secretion of formed and new synthesized mediators. Mast cells have more than 30 known surface receptors that activate different pathways for direct and indirect activation by microbes. Different bacterial strains stimulate mast cells through various ligands, initiating the innate immune response, which aids in clearing the bacterial burden. Mast cell interactions with adaptative immune cells also play a crucial role in infections. Recent publications revealed another "non-canonical" degranulation mechanism present in tryptase and chymase mast cells in humans and connective tissue mast cells in mice, occurring through the activation of the Mas-related G protein-coupled receptor (MRGPRX2/b2). This receptor represents a new therapeutic target alongside antibiotic therapy. There is an urgent need to reconsider and redefine the biological role of these MASTer cells of innate immunity, extending beyond their involvement in allergic pathology.
Assuntos
Anti-Infecciosos , Hipersensibilidade , Humanos , Animais , Camundongos , Anti-Infecciosos/metabolismo , Citocinas/metabolismo , Imunoglobulina E , Imunidade Inata , Mastócitos , Proteínas do Tecido Nervoso/metabolismo , Receptores de Neuropeptídeos/metabolismo , Receptores Acoplados a Proteínas G/metabolismoRESUMO
SUMMARY: Mast cells (MC) are cells of the immune system that regulate cell and tissue homeostasis, are found in low numbers, have an intact plasma membrane, and a cytoplasm with a wide variety of inflammatory chemical mediators. The activation or degranulation of mast cells implies the release of these chemical mediators (interleukins, cytokines, and more), causing tissue actions ranging from the activation of metalloproteinases to the development of anaphylactic hypersensitivity of different degrees, alterations in vascular permeability, and loss of cell homeostasis. This behavior would allow them to act as sentinels responding to pathophysiological processes. During the COVID-19 pandemic, in positive human patients, the available literature reports the presence and degranulation of mast cells in a generalized manner, especially in the respiratory tract. This study aimed to analyze the emerging role of MCs in the pathogenesis of diseases and their projection as biological markers in the treatment of diseases or pandemics. The analysis of human biopsies showed that MCs are observed as cells with diameters between 8 to 20 µm, and in inflamed tissues, degranulation of MCs is observed. The action of MCs degranulation was related to different inflammatory processes of autoimmune diseases. It is concluded that the potential of MC as therapeutic targets and biomarkers could raise new pharmacological targets, as supportive therapy, and possibly of great help in the treatment of future emerging pandemics such as the current monkeypox.
Los mastocitos (MC) son células del sistema inmune que regulan la homeostasis celular y tisular, se encuentran en escasas cantidades, presentan una membrana plasmática íntegra, y un citoplasma con una amplia variedad de mediadores químicos. La activación o degranulación de los mastocitos implica la liberación de estos mediadores químicos (interleuquinas, citoquina y más), provocando acciones tisulares que van desde la activación de metaloproteinasas hasta el desarrollo de hipersensibilidad anafiláctica de distinto grado, provocando la pérdida de la homeostasis celular. Durante la pandemia de la COVID-19, en pacientes humanos positivos, se informa recurrentemente la presencia y degranulación de mastocitos de manera generalizada sobre todo en las vías respiratorias. El análisis de la degranulación de los MCs podría proporcionar información que podría utilizarse en el desarrollo de tratamientos preventivos contra infecciones virales, bacterianas u otros patógenos. Este comportamiento les permitiría actuar como centinelas en respuesta a procesos fisiopatológicos. El objetivo de este trabajo fue analizar el rol emergente de los MCs en la patogenia de enfermedades y su proyección como marcadores biológicos en el tratamiento de enfermedades o pandemias. En análisis de biopsias humanas se muestran que MCs se observan como células con diámetros de entre 8 a 20 µm, en tejidos inflamados se observa degranulación de MCs. Se relacionó el accionar de degranulación de los MCs en diferentes procesos inflamatorios de enfermedades autoinmunes. Se concluye que el potencial de MC como dianas terapéuticas y biomarcadores podrían plantear nuevos objetivos farmacológicos, como terapia de apoyo, y posiblemente de gran ayuda en el tratamiento de futuras pandemias emergentes como la actual viruela del mono.
Assuntos
Humanos , Controle de Doenças Transmissíveis/métodos , Doenças Transmissíveis/patologia , Mastócitos , Biomarcadores , Saúde Pública , Doenças Transmissíveis/imunologia , Emergências , Pandemias/prevenção & controle , COVID-19/imunologia , COVID-19/patologia , COVID-19/prevenção & controleRESUMO
Maternal malnutrition can alter developmental biology, programming health and disease in offspring. The increase in sugar consumption during the peripubertal period, a worldwide concern, also affects health through adulthood. Studies have shown that maternal exposure to a low protein diet (LPD) is associated with an increase in prostate disease with aging. However, the combined effects of maternal LPD and early postnatal sugar consumption on offspring prostate disorders were not investigated. The effects on aging were evaluated using a maternal gestational model with lactational LPD (6% protein) and sugar consumption (10%) from postnatal day (PND) 21-90, associating the consequences on ventral prostate (VP) rats morphophysiology on PND540. An increase was shown in mast cells and in the VP of the CTR + SUG and Gestational and Lactational Low Protein (GLLP) groups. In GLLP + SUG, a significant increase was shown in TGF-ß1 expression in both the systemic and intra-prostatic forms, and SMAD2/3p had increased. The study identified maternal LPD and sugar consumption as risk factors for prostatic homeostasis in senility, activating the TGFß1-SMAD2/3 pathway, a signaling pathway with potential markers for prostatic disorders.
Assuntos
Desnutrição , Fenômenos Fisiológicos da Nutrição Materna , Efeitos Tardios da Exposição Pré-Natal , Próstata , Doenças Prostáticas , Animais , Masculino , Feminino , Gravidez , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Doenças Prostáticas/patologia , Doenças Prostáticas/etiologia , Doenças Prostáticas/metabolismo , Desnutrição/complicações , Próstata/metabolismo , Próstata/patologia , Ratos , Inflamação/patologia , Inflamação/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Fator de Crescimento Transformador beta1/genética , Dieta com Restrição de Proteínas/efeitos adversos , Proteína Smad2/metabolismo , Ratos Wistar , Proteína Smad3/metabolismo , Proteína Smad3/genética , Transdução de Sinais , Animais Recém-Nascidos , Mastócitos/metabolismoRESUMO
Methadone is an effective and long-lasting analgesic drug that is also used in medication-assisted treatment for people with opioid use disorders. Although there is evidence that methadone activates µ-opioid and Toll-like-4 receptors (TLR-4s), its effects on distinct immune cells, including mast cells (MCs), are not well characterized. MCs express µ-opioid and Toll-like receptors (TLRs) and constitute an important cell lineage involved in allergy and effective innate immunity responses. In the present study, murine bone-marrow-derived mast cells (BMMCs) were treated with methadone to evaluate cell viability by flow cytometry, cell morphology with immunofluorescence and scanning electron microscopy, reactive oxygen species (ROS) production, and intracellular calcium concentration ([Ca2+]i) increase. We found that exposure of BMMCs to 0.5 mM or 1 mM methadone rapidly induced cell death by forming extracellular DNA traps (ETosis). Methadone-induced cell death depended on ROS formation and [Ca2+]i. Using pharmacological approaches and TLR4-defective BMMC cultures, we found that µ-opioid receptors were necessary for both methadone-induced ROS production and intracellular calcium increase. Remarkably, TLR4 receptors were also involved in methadone-induced ROS production as it did not occur in BMMCs obtained from TLR4-deficient mice. Finally, confocal microscopy images showed a significant co-localization of µ-opioid and TLR4 receptors that increased after methadone treatment. Our results suggest that methadone produces MCETosis by a mechanism requiring a novel crosstalk pathway between µ-opioid and TLR4 receptors.
Assuntos
Analgésicos Opioides , Armadilhas Extracelulares , Humanos , Animais , Camundongos , Analgésicos Opioides/farmacologia , Receptor 4 Toll-Like/metabolismo , Metadona/farmacologia , Mastócitos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Medula Óssea/metabolismo , Cálcio/metabolismo , Armadilhas Extracelulares/metabolismo , Receptor 2 Toll-Like/metabolismo , Receptores Toll-Like/metabolismoRESUMO
Fibromyalgia is a painful disorder of unknown aetiology that presents activation and recruitment of innate immune cells, including mast cells. Efforts have been made to understand its pathogenesis to manage it better. Thus, we explored the involvement of peripheral mast cells in an experimental model of fibromyalgia induced by reserpine. Reserpine (1 mg/kg) was subcutaneously (s.c.) injected once daily in the back of male Swiss mice for three consecutive days. We analysed mechanical and cold allodynia, muscle fatigue and number of mast cell in plantar tissue. The fibromyalgia induction produced mast cell infiltration (i.e., mastocytosis) in the mice's plantar tissue. The depletion of mast cell mediators with the compound 48/80 (0.5-4 mg/kg, intraperitoneal (i.p.)) or the mast cell membrane stabilizer ketotifen fumarate (10 mg/kg, oral route (p.o.) widely (80-90 %) and extensively (from 1 up to 10 days) prevented reserpine-induced mechanical and cold allodynia and muscle fatigue. Compound 48/80 also prevented the reserpine-induced mastocytosis. Finally, we demonstrated that PAR-2, 5-HT2A, 5-HT3, H1, NK1 and MrgprB2 receptors, expressed in neuronal or mast cells, seem crucial to mediate fibromyalgia-related cardinal symptoms since antagonists or inhibitors of these receptors (gabexate (10 mg/kg, s.c.), ENMD-1068 (10 mg/kg, i.p.), ketanserin (1 mg/kg, i.p.), ondansetron (1 mg/kg, p.o.), promethazine (1 mg/kg, i.p.), and L733,060 (5 mg/kg, s.c.), respectively) transiently reversed the reserpine-induced allodynia and fatigue. The results indicate that mast cells mediate painful and fatigue behaviours in this fibromyalgia model, representing potential therapy targets to treat fibromyalgia syndrome.
Assuntos
Fibromialgia , Mastocitose , Camundongos , Masculino , Animais , Fibromialgia/metabolismo , Mastócitos/metabolismo , Hiperalgesia/metabolismo , Serotonina/metabolismo , Reserpina/efeitos adversos , Mastocitose/metabolismo , Mastocitose/patologiaRESUMO
The aim of this study was to analyze the expression of mast cell markers toluidine blue, c-kit, and tryptase and presence of mononuclear inflammatory cells in oral lichen planus (OLP) and oral lichenoid lesions related to dental amalgam. Nineteen specimens of OLP, OLLC, and healthy oral mucosa were selected. Mononuclear inflammatory cells were analyzed. Histochemical and immunohistochemical analyses were performed using toluidine blue, anti-c-kit and anti-tryptase reagents, and the results were quantified in areas A and B of connective tissue. Mast cells of all OLP and OLLC samples were positive for toluidine blue, c-kit, and tryptase. The density of toluidine blue+, c-kit+ and tryptase+ mast cells was higher in tissue with OLP and OLLC compared with healthy controls (p < 0.05). No difference was noted in mast cells density between OLP and OLLC (p > 0.05). The density of tryptase+ mast cells was higher in the subepithelial region (area A) than the region below it (Area B) in OLLC (p = 0.047). The mononuclear inflammatory cell density was higher in OLLC compared to OLP, but without statistical significance (p > 0.05). A positive statistical correlation was found between mononuclear immune cells and density of c-kit+ and tryptase+ mast cells in OLP (r = 0.943 and r = 0.886, respectively). Our data demonstrate that the etiopathogenesis process of OLP and OLLC modulates the expansion and degranulation of mast cells; mast cells density, however, was similar between OLP and OLLC. The distribution of mast cells appears to vary along the lamina propria.
Assuntos
Líquen Plano Bucal , Mastócitos , Humanos , Amálgama Dentário/efeitos adversos , Cloreto de Tolônio/efeitos adversos , TriptasesRESUMO
Increased or constitutive activation of nuclear factor kappa B (NF-kB) is a feature of many chronic disease processes, including cancer. While NF-kB overactivation has been documented extensively in human oncology, there is a relative paucity of data documenting the same phenomenon in veterinary medicine. To assess NF-kB activity, antibodies to p65 and p100/p52, which are components of NF-kB heterodimers, were first validated for specificity and canine cross-reactivity via Western blot and labeling of immortalized cell pellets. Then, nuclear labeling for these antibodies was assessed via QuPath software in over 200 tumor tissue samples (10 hemangiosarcomas, 94 histiocytic sarcomas, 71 lymphomas, and 28 mast cell tumors) and compared to immunolabeling in appropriate normal tissue counterparts. Greater than 70% of spontaneous canine tumors evaluated in this study had more nuclear p65 and p100/p52 immunoreactivity than was observed in comparable normal cell populations. Specifically, 144/204 (70.58%) of tumors evaluated had positive p65 nuclear labeling and 179/195 (91.79%) had positive p100/p52 nuclear labeling. Surprisingly, greater nuclear p100/p52 reactivity was associated with a longer progression-free survival (PFS) and overall survival (OS) in canine lymphomas. These results provide support and preliminary data to investigate the role of NF-kB signaling in different types of canine cancer.
Assuntos
Doenças do Cão , Hemangiossarcoma , Sarcoma Histiocítico , Linfoma , Animais , Cães , Humanos , NF-kappa B/metabolismo , Sarcoma Histiocítico/veterinária , Hemangiossarcoma/veterinária , Mastócitos , Subunidade p52 de NF-kappa B/metabolismo , Linfoma/veterináriaAssuntos
Mastocitose Sistêmica , Mastocitose , Humanos , Mastócitos/metabolismo , Mastócitos/patologia , Brasil , Centros de Atenção Terciária , Mastocitose/diagnóstico , Mastocitose/metabolismo , Mastocitose/patologia , Biópsia , Algoritmos , Mastocitose Sistêmica/diagnóstico , Proteínas Proto-Oncogênicas c-kitRESUMO
OBJECTIVE: The aim of this study was to compare, in adults and elderly individuals, the immunoexpression of immature and mature dendritic cells (DCs), mast cells, and blood vessels in healthy and diseased gingival tissues. MATERIALS AND METHODS: The expressions of immunohistochemical markers, including CD1a (immature dendritic cells), CD83 (mature dendritic cells), tryptase (mast cells) and CD34 (blood vessels), were analyzed in gingival biopsies from elderly (n = 27) and adult (n = 127) patients presenting health, gingivitis and periodontitis. Positive cells for each specimen and marker were counted. RESULTS: There were no differences in the immunostaining of DCs, mast cells and the amount of blood vessels among gingival biopsies with health, gingivitis and periodontitis in adult and elderly subjects (p > 0.05). Immature DCs were more frequent in tissues with gingivitis and periodontitis in elderly patients, when compared to adults (p < 0.05). Furthermore, degranulated mast cell counts were higher, whereas the number of microvessels was lower in gingivitis in the elderly, when compared to adults (p < 0.05). CONCLUSIONS: Diseased periodontal sites in the elderly present an overall significant overexpression of immature DCs and degranulated mast cells, in relation to those of adults. Furthermore, gingivitis in elderly is associated with decreased microvessel growth. These immunoinflammatory differences between elderly and adults may have implications in periodontal tissue breakdown in the late adulthood. Further studies should be performed to elucidate this hypothesis. CLINICAL RELEVANCE: Understading the relationship between aging and changes in immune cells during periodontal inflammation may lead to therapeutic targets for the future management of periodontal diseases.