RESUMEN
This study evaluated the role of kinin B1 and B2 receptors in the pre-clinical mouse model of oxazolone-induced atopic dermatitis. The B1 R715 or B2 HOE140 receptor antagonists were dosed at different schemes of treatment. After assessment of clinical lesion scores and pruritus, lesional skin samples were collected for histopathological analysis. The plasma extravasation and the expression of the metalloproteinase ADAMTS5 were also assessed. The immunopositivity for kinin receptors was evaluated in the skin, dorsal root ganglion (DRG), thoracic spinal cord and brain cortex sections. Marked upregulation of B1 and B2 receptors was observed in the skin of oxazolone-treated mice. The induction of atopic dermatitis led to a downregulation of both receptors in the DRG, without any alteration in the spinal cord and brain cortex. The repeated administration of HOE140 (50â¯nmol/kg; i.p.) partially inhibited the oxazolone-related pruritus, associated with a reduction of ADAMTS5 immunolabelling in the skin. Alternatively, R715 (438â¯nmol/kg; i.p.) produced a mild inhibition of plasma extravasation in oxazolone-challenged mice. Noteworthy, the repeated i.d. injection of R715 (30â¯nmol/site) or HOE140 (3â¯nmol/site) significantly reduced the histiocyte numbers, according to the histopathological analysis. Either B1 or B2 kinin antagonists, irrespective of the protocol of treatment, did not alter any other evaluated clinical or histological parameters. Data brings novel evidence about the role of kinin receptors in allergy-related conditions, such as atopic dermatitis. Further studies to test different protocols of treatment with kinin antagonists on in-depth cellular alterations underlying oxazolone-induced atopic dermatitis remain to be performed.
Asunto(s)
Dermatitis Atópica/inmunología , Receptor de Bradiquinina B1/inmunología , Receptor de Bradiquinina B2/inmunología , Animales , Corteza Cerebral/metabolismo , Dermatitis Atópica/inducido químicamente , Dermatitis Atópica/metabolismo , Modelos Animales de Enfermedad , Ganglios Espinales/metabolismo , Masculino , Ratones , Oxazolona , Receptor de Bradiquinina B1/metabolismo , Receptor de Bradiquinina B2/metabolismo , Médula Espinal/metabolismoRESUMEN
The role, if any, played by the kinin system in tuberculosis infection models, either in vivo or in vitro, was investigated. The effects of Mycobacterium tuberculosis infection on C57BL/6 wild type, B1R-/-, B2R-/- and double B1R/B2R knockout mice were evaluated. Immunohistochemistry analysis was carried out to assess B1R and B2R expression in spleens and lungs of M. tuberculosis-infected mice. In addition, in vitro experiments with M. tuberculosis-infected macrophages were performed. The in vivo effects of HOE-140 and SSR240612 on the mice model of infection were also evaluated. Infected B2R-/- mice exhibited increased splenomegaly, whereas decreased spleen weight in infected double B1R/B2R knockout mice was observed. The bacterial load, determined as colony-forming units, did not differ in the spleens and lungs of the studied mouse strains. Importantly, immunohistochemical analysis revealed that B1R was upregulated in both spleens and lungs of infected mice. M. tuberculosis-infected macrophages incubated with SSR240612, alone or in combination with des-Arg9-BK, for four days, displayed a marked inhibitory effect on CFU counts. However, the pre-incubation of the selective B1R (des-Arg9-BK and SSR240612) and B2R (BK and HOE-140) agonists and antagonists, respectively, did not significantly affect the bacterial loads. A statistically significant reduction in the CFU of M. tuberculosis in lungs and spleens of animals treated with SSR240612, but not with HOE-140, was observed. Further efforts should be pursued to clarify whether or not SSR240612 might be considered an option for the treatment of tuberculosis.
Asunto(s)
Antituberculosos/administración & dosificación , Antagonistas del Receptor de Bradiquinina B1/administración & dosificación , Dioxoles/administración & dosificación , Pulmón/efectos de los fármacos , Mycobacterium tuberculosis/efectos de los fármacos , Receptor de Bradiquinina B1/efectos de los fármacos , Sulfonamidas/administración & dosificación , Tuberculosis Pulmonar/tratamiento farmacológico , Administración Oral , Animales , Carga Bacteriana , Bradiquinina/administración & dosificación , Bradiquinina/análogos & derivados , Antagonistas del Receptor de Bradiquinina B2/administración & dosificación , Modelos Animales de Enfermedad , Femenino , Pulmón/metabolismo , Pulmón/microbiología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Macrófagos/microbiología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Mycobacterium tuberculosis/crecimiento & desarrollo , Células RAW 264.7 , Receptor de Bradiquinina B1/deficiencia , Receptor de Bradiquinina B1/genética , Receptor de Bradiquinina B1/metabolismo , Receptor de Bradiquinina B2/genética , Receptor de Bradiquinina B2/metabolismo , Bazo/efectos de los fármacos , Bazo/metabolismo , Bazo/microbiología , Tuberculosis Pulmonar/genética , Tuberculosis Pulmonar/metabolismo , Tuberculosis Pulmonar/microbiologíaRESUMEN
Mephedrone and methedrone are cathinone-related compounds, which act as non-selective substrates for monoamine transporters, facilitating a neurotransmitter release. We compared the acute pharmacological effects of mephedrone and methedrone, attempting to further evaluate the action mechanisms of methedrone by responsibly and ethically using mice under approved procedures. The effects of both compounds were examined from 10 to 60 min, in a series of behavioral paradigms, namely open-field, plus-maze, hot-plate and tail suspension tests, whereas neurotransmitter brain tissue levels were determined ex vivo by HPLC. Separate groups were pre-treated with the dopamine (DA) antagonist haloperidol, or the serotonin (5-HT) synthesis inhibitor ρCPA, to further assess the mechanisms underlying methedrone effects. The compounds caused marked hyperlocomotion, displaying dissimilar stereotyped behavior, in an open-field arena. Mephedrone caused anxiolytic-like effects, while methedrone induced anxiogenic-like actions in the elevated plus-maze. Both compounds displayed thermal antinociception, with a reduced immobility time in the tail suspension model. Mephedrone triggered a 2- and 3-fold increment of dopamine and serotonin tissue levels, respectively, in the nucleus accumbens, with a 1.5-fold elevation of tissue dopamine in the frontal cortex. Methedrone caused a 2-fold increment of tissue dopamine in the nucleus accumbens and in the striatum, and a 1.5-fold increment of serotonin tissue levels in the hippocampus and striatum. In vivo methedrone effects were partially inhibited by a pre-treatment with haloperidol or ρCPA. Despite similar actions on locomotion, analgesia, and depression-like behavior, the acute administration of mephedrone and methedrone elicited divergent effects on anxiety-like behavior and stereotyped movements in mice, which might be related to the distinct modulation of brain tissue neurotransmitter levels.