RESUMO
Glucocerebrosidase gene (GBA) mutations are the most common genetic contributor to Parkinson's disease (PD) and are associated with decreased glucocerebrosidase (GCase) enzymatic activity in PD. PD patients without GBA mutations also exhibit lower levels of GCase activity in the central nervous system suggesting a potential contribution of the enzyme activity in disease pathogenesis, possibly by alteration of lysosomal function. α-synuclein (ASYN), a protein with a central role in PD pathogenesis, has been shown to be secreted partly in association with exosomes. It is possible that a dysfunction of the endocytic pathway through GCase may result in altered exosome release of ASYN. The aim of this study was to examine whether manipulating GCase activity in vivo and in vitro could affect ASYN accumulation and secretion. GCase overexpression in vitro resulted in a significant decrease of exosome secretion. Chronic inhibition of GCase activity in vivo, by administration of the covalent inhibitor conduritol-B epoxide in A53T-synuclein alpha gene Tg mice significantly elevated intracellular oligomeric ASYN species. Importantly, GCase inhibition, induced a profound increase in the number of brain exosomes released, as well as exosome-associated ASYN oligomers. Finally, virus-mediated expression of mutant GBA in the mouse striatum increased ASYN secretion in the same region. Together, these results provide for the first time evidence that a decrease of GCase or overexpression of mutant GCase in a chronic in vivo setting can affect ASYN secretion. Such effects may mediate enhanced propagation of ASYN, driving pathology in GBA-associated PD.
Assuntos
Exossomos/genética , Glucosilceramidase/genética , Doença de Parkinson/genética , alfa-Sinucleína/genética , Animais , Encéfalo/metabolismo , Encéfalo/fisiopatologia , Modelos Animais de Doenças , Exossomos/metabolismo , Glucosilceramidase/metabolismo , Humanos , Inositol/administração & dosagem , Inositol/análogos & derivados , Lisossomos/genética , Lisossomos/metabolismo , Camundongos , Mutação , Doença de Parkinson/fisiopatologiaRESUMO
Hydrogen sulfide (H2S) is an endogenous gasotransmitter with anti-inflammatory actions that also reduces itching. To test whether a combination of an antihistamine with a H2S donor has improved antipruritic efficacy, bifunctional molecules with antihistamine and H2S-releasing pharmacophores were synthesized and tested in vitro and in vivo. H2S release from the hybrid molecules was evaluated with the methylene blue and lead acetate methods, and H1-blocking activity was assessed by determining tissue factor expression inhibition. All new compounds released H2S in a dose-dependent manner and retained histamine blocking activity. Two compounds with the highest potency were evaluated in vivo for their antipruritic as well as sedative action; they proved to possess higher efficacy in inhibiting histamine-induced pruritus and decreased sedative effects compared to the parent compounds (hydroxyzine and cetirizine), suggesting that they exhibit superior antipruritic action and limited side effects that likely arise from the H2S-releasing moiety.
Assuntos
Antipruriginosos , Sulfeto de Hidrogênio , Humanos , Antipruriginosos/uso terapêutico , Hipnóticos e Sedativos/farmacologia , Hipnóticos e Sedativos/uso terapêutico , Histamina , Antagonistas dos Receptores Histamínicos H1/farmacologia , Antagonistas dos Receptores Histamínicos H1/uso terapêutico , Antagonistas dos Receptores Histamínicos/farmacologia , Antagonistas dos Receptores Histamínicos/uso terapêutico , Prurido/tratamento farmacológico , Sulfeto de Hidrogênio/farmacologia , Sulfeto de Hidrogênio/uso terapêuticoRESUMO
PURPOSE: Glioblastoma (GBM) is the most common type of adult brain tumor with extremely poor survival. Cystathionine-gamma lyase (CTH) is one of the main Hydrogen Sulfide (H2S) producing enzymes and its expression contributes to tumorigenesis and angiogenesis but its role in glioblastoma development remains poorly understood. METHODS: and Principal Results: An established allogenic immunocompetent in vivo GBM model was used in C57BL/6J WT and CTH KO mice where the tumor volume and tumor microvessel density were blindly measured by stereological analysis. Tumor macrophage and stemness markers were measured by blinded immunohistochemistry. Mouse and human GBM cell lines were used for cell-based analyses. In human gliomas, the CTH expression was analyzed by bioinformatic analysis on different databases. In vivo, the genetic ablation of CTH in the host led to a significant reduction of the tumor volume and the protumorigenic and stemness transcription factor sex determining region Y-box 2 (SOX2). The tumor microvessel density (indicative of angiogenesis) and the expression levels of peritumoral macrophages showed no significant changes between the two genotypes. Bioinformatic analysis in human glioma tumors revealed that higher CTH expression is positively correlated to SOX2 expression and associated with worse overall survival in all grades of gliomas. Patients not responding to temozolomide have also higher CTH expression. In mouse or human GBM cells, pharmacological inhibition (PAG) or CTH knockdown (siRNA) attenuates GBM cell proliferation, migration and stem cell formation frequency. MAJOR CONCLUSIONS: Inhibition of CTH could be a new promising target against glioblastoma formation.
Assuntos
Glioblastoma , Camundongos , Humanos , Animais , Glioblastoma/tratamento farmacológico , Glioblastoma/genética , Glioblastoma/patologia , Cistationina gama-Liase/genética , Cistationina gama-Liase/metabolismo , Camundongos Endogâmicos C57BL , Temozolomida , Linhagem Celular , Linhagem Celular TumoralRESUMO
Neurogenesis in the dentate gyrus (DG) of the adult hippocampus is actively involved in brain homeostasis. Thus, identification of novel regulators in adult neurogenesis could significantly contribute to new therapies. We have recently unraveled the regulatory role of NR5A2 (also known as LRH1), a druggable orphan nuclear receptor, in embryonic neurogenesis. However, its involvement in adult neurogenesis is still an open question. Here we show that NR5A2 is differentially expressed in the DG of the adult hippocampus with neurons exhibiting higher levels of expression than adult neural stem/progenitor cells (aNSCs), suggesting a correlation with neuronal differentiation. Notably, NR5A2 overexpression in ex vivo cultured aNSCs induces expression of Prox1, a critical regulator of adult hippocampal neurogenesis. In agreement, NR5A2 is sufficient to reduce proliferation, increase neuronal differentiation, and promote axon outgrowth. Moreover, depletion of NR5A2 in DG cells in vivo caused a decrease in the number of NeuN as well as Calbindin-positive neurons, indicating its necessity for the maintenance of neuronal identity. Our data propose a regulatory role of NR5A2 in neuronal differentiation and fate specification of adult hippocampal NSCs.
Assuntos
Hipocampo/metabolismo , Células-Tronco Neurais/metabolismo , Neurogênese/fisiologia , Neurônios/metabolismo , Animais , Calbindinas/metabolismo , Ciclo Celular/fisiologia , Proliferação de Células/fisiologia , CamundongosRESUMO
INTRODUCTION: Psoriasis, is a common, chronic, autoimmune, inflammatory, relapsing disease, which would benefit from reliable and human-relevant animal models to test drugs pre-clinically and to understand their mechanism of action. Because of its ease of use, convenience and low cost, the imiquimod (IMQ)-induced psoriasis-like model is widely utilized; however, it is not known whether all mouse strains are equivalent and if the hairless mouse is appropriate, so that the imiquimod model can be further optimized. METHODS: Under similar experimental conditions, common mouse strains (BALB/c, C57BL/6J, and ApoE) and a new hairless strain (ApoE/SKH-hr2) as well as several inducers (IMQ, IMQ + acetic acid (AcOH) topical and IMQ + AcOH systemic) were compared by clinical, histopathological, biophysical and locomotor activity assessments. RESULTS AND DISCUSSION: The BALB/c mice yielded an optimal psoriasis-like phenotype with IMQ + AcOH topical treatment, and the corresponding phenotypes for the other mouse strains were C57BL/6J moderate and ApoE mild. In contrast, the ApoE/SKH-hr2 mice, as a result of the absence of a Munro abscess in the histopathology analysis, left doubt about the psoriasis-like acquisition. Locomotor activity of BALB/c mice treated with IMQ, IMQ + AcOH topically and IMQ + AcOH systemically showed decreased distance and rearing coverage and increased immobility with all treatments. Hence, the BALB/c mouse strain appears to be an optimal psoriasis-like model when utilizing IMQ + AcOH topical application.
Assuntos
Psoríase , Animais , Modelos Animais de Doenças , Imiquimode/toxicidade , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Psoríase/induzido quimicamente , Psoríase/tratamento farmacológico , PeleRESUMO
Chronic wounds unresponsive to existing treatments constitute a serious disease burden. Factors that contribute to the pathogenesis of chronic ulcers include oxidative stress, comorbid microbial infections, and the type of immune system response. Preclinically, and in a case study, a formulation containing a Ceratothoa oestroides olive oil extract promoted wound healing. Patients with chronic venous and pressure ulcers, clinically assessed as being unresponsive to healing agents, were treated for 3 months with an ointment containing the C oestroides extract combined with antibiotic and/or antiseptic agents chosen according to the type of bacterial infection. Treatment evaluation was performed using the Bates-Jensen criteria with +WoundDesk and MOWA cell phone applications. After 3 months of treatment, C oestroides resulted in an average decrease of 36% in the Bates-Jensen score of ulcers (P < .000), with the decrease being significant from the first month (P < .007). The combined use of topically applied antibiotics and antiseptics efficiently controlled microbial ulcer infection and facilitated wound healing. In relation to other factors such as initial wound size, chronicity appeared to be an important prognostic factor regarding the extent of wound healing. Future clinical investigations assessing the wound healing efficacy of the C oestroides olive oil extract are warranted.
Assuntos
Isópodes/química , Azeite de Oliva/administração & dosagem , Úlcera por Pressão , Extratos de Tecidos/administração & dosagem , Úlcera Varicosa , Infecção dos Ferimentos , Idoso , Animais , Antibacterianos/administração & dosagem , Anti-Infecciosos Locais/administração & dosagem , Bactérias/citologia , Bactérias/efeitos dos fármacos , Bactérias/isolamento & purificação , Curativos Biológicos , Feminino , Humanos , Masculino , Avaliação de Resultados em Cuidados de Saúde/métodos , Úlcera por Pressão/diagnóstico , Úlcera por Pressão/fisiopatologia , Úlcera por Pressão/terapia , Úlcera Varicosa/diagnóstico , Úlcera Varicosa/fisiopatologia , Úlcera Varicosa/terapia , Cicatrização/efeitos dos fármacos , Infecção dos Ferimentos/microbiologia , Infecção dos Ferimentos/terapiaRESUMO
Lithium is the most effective mood stabilizer for the treatment of bipolar disorder, but it is toxic at only twice the therapeutic dosage and has many undesirable side effects. It is likely that a small molecule could be found with lithium-like efficacy but without toxicity through target-based drug discovery; however, therapeutic target of lithium remains equivocal. Inositol monophosphatase is a possible target but no bioavailable inhibitors exist. Here we report that the antioxidant ebselen inhibits inositol monophosphatase and induces lithium-like effects on mouse behaviour, which are reversed with inositol, consistent with a mechanism involving inhibition of inositol recycling. Ebselen is part of the National Institutes of Health Clinical Collection, a chemical library of bioavailable drugs considered clinically safe but without proven use. Therefore, ebselen represents a lithium mimetic with the potential both to validate inositol monophosphatase inhibition as a treatment for bipolar disorder and to serve as a treatment itself.