RESUMO
Silicosis is a diffuse fibrotic lung disease in which excessive inflammatory responses are triggered by silica exposure. Pyroptosis, a pro-inflammatory mode of programmed cell death, is mediated by gasdermin and may play a pivotal role in the development of silicosis. The caspase-1 inhibitor, VX-765, was used in vivo and in vitro to investigate the effects of silica-induced early inflammatory injury and later lung fibrosis. Our findings show that VX-765 reduces inflammatory lung injury by inhibiting silica-induced pyroptosis of alveolar macrophages in a silicosis mouse model. VX-765 limits the infiltration of inflammatory M1 alveolar macrophages, decreasing expression of inflammatory cytokines, including IL-1ß, TNF-α, IL-6, CCL2, and CCL3, and down-regulating endogenous DAMPs and inflammatory immune-related cell pattern recognition receptors TLR4 and NLRP3. Furthermore, VX-765 alleviates fibrosis by down-regulating α-smooth muscle actin (α-SMA), collagen, and fibronectin. In this study, we illustrate that Alveolar macrophages pyroptosis occur in the early stages of silicosis, and VX-765 can alleviate the development of silicosis by inhibiting the pyroptosis signaling pathway. These results may provide new insight into the prevention and treatment of early-stage silicosis.
Assuntos
Inibidores de Caspase , Lesão Pulmonar , Fibrose Pulmonar , Piroptose , Silicose , Animais , Camundongos , Lesão Pulmonar/induzido quimicamente , Lesão Pulmonar/tratamento farmacológico , Lesão Pulmonar/patologia , Macrófagos Alveolares/efeitos dos fármacos , Piroptose/efeitos dos fármacos , Dióxido de Silício/toxicidade , Silicose/tratamento farmacológico , Inibidores de Caspase/farmacologia , Inibidores de Caspase/uso terapêutico , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/tratamento farmacológicoRESUMO
Ulcerative colitis (UC) is a chronic inflammatory bowel disease. Several studies have demonstrated that α7 nicotinic acetylcholine receptors (α7nAChRs) exert anti-inflammatory effects on immune cells and nicotine suppress UC onset and relapse. Plasmacytoid dendritic cells (pDCs) reportedly accumulate in the colon of UC patients. Therefore, we investigated the pathophysiological roles of α7nAChRs on pDCs in the pathology of UC using oxazolone (OXZ)-induced Th2-type colitis with BALB/c mice. 2-deoxy-D-glucose, a central vagal stimulant suppressed OXZ colitis, and nicotine also ameliorated OXZ colitis with suppressing Th2 cytokines, which was reversed by α7nAChR antagonist methyllycaconitine. Additionally, α7nAChRs were expressed on pDCs, which were located very close to cholinergic nerve fibers in the colon of OXZ mice. Furthermore, nicotine suppressed CCL21-induced bone marrow-derived pDC migration due to Rac 1 inactivation, which was reversed by methyllycaconitine, a JAK2 inhibitor AG490 or caspase-3 inhibitor AZ-10417808. CCL21 was mainly expressed in the isolated lymphoid follicles (ILFs) of the colon during OXZ colitis. The therapeutic effect of cholinergic pathway on OXZ colitis probably through α7nAChRs on pDCs were attributed to the suppression of pDC migration toward the ILFs. Therefore, the activation of α7nAChRs has innovative therapeutic potential for the treatment of UC.
Assuntos
Neurônios Colinérgicos/efeitos dos fármacos , Colite Ulcerativa/tratamento farmacológico , Células Dendríticas/efeitos dos fármacos , Neuroimunomodulação , Células Th2/metabolismo , Aconitina/análogos & derivados , Aconitina/farmacologia , Aconitina/uso terapêutico , Animais , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Caspase 3/metabolismo , Inibidores de Caspase/farmacologia , Inibidores de Caspase/uso terapêutico , Estimulantes do Sistema Nervoso Central/farmacologia , Estimulantes do Sistema Nervoso Central/uso terapêutico , Colite Ulcerativa/induzido quimicamente , Colo/metabolismo , Células Dendríticas/metabolismo , Desoxiglucose/farmacologia , Desoxiglucose/uso terapêutico , Modelos Animais de Doenças , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/uso terapêutico , Janus Quinase 2/metabolismo , Camundongos Endogâmicos BALB C , Neuropeptídeos/metabolismo , Nicotina/farmacologia , Nicotina/uso terapêutico , Oxazolona/toxicidade , Fator de Transcrição STAT3/metabolismo , Células Th2/efeitos dos fármacos , Tirfostinas/farmacologia , Tirfostinas/uso terapêutico , Nervo Vago/efeitos dos fármacos , Receptor Nicotínico de Acetilcolina alfa7/agonistas , Receptor Nicotínico de Acetilcolina alfa7/antagonistas & inibidores , Receptor Nicotínico de Acetilcolina alfa7/genética , Receptor Nicotínico de Acetilcolina alfa7/metabolismo , Proteínas rac1 de Ligação ao GTP/metabolismoRESUMO
BACKGROUND: Multiple studies have revealed that repeated or long-term exposure to ketamine causes neurodegeneration and cognitive dysfunction. Pyroptosis is an inflammatory form of programmed cell death that has been linked to various neurological diseases. However, the role of NLRP3/caspase-1 axis-related pyroptosis in ketamine-induced neurotoxicity and cognitive dysfunction remains uncertain. METHODS: To evaluate whether ketamine caused NLRP3/caspase1-dependent pyroptosis, flow cytometry analysis, western blotting, ELISA test, histopathological analysis, Morris water maze (MWM) test, cell viability assay, and lactate dehydrogenase release (LDH) assay were carried out on PC12 cells, HAPI cells, and 7-day-old rats. In addition, the NLRP3 inhibitor MCC950 or the caspase-1 inhibitor VX-765 was used to investigate the role of the NLRP3/caspase-1 axis in ketamine-induced neurotoxicity and cognitive dysfunction. RESULTS: Our findings demonstrated that ketamine exposure caused cell damage and increased the levels of pyroptosis in PC12 cells, HAPI cells, and the hippocampus of neonatal rats. After continuous exposure to ketamine, targeting NLRP3 and caspase-1 with MCC950 or VX765 improved pyroptosis, reduced neuropathological damages, and alleviated cognitive dysfunction. CONCLUSION: NLRP3/Caspase-1 axis-dependent pyroptosis is involved in ketamine-induced neuroinflammation and cognitive dysfunction, and it provides a promising strategy to treat ketamine-related neurotoxicity.
Assuntos
Caspase 1/metabolismo , Disfunção Cognitiva/metabolismo , Hipocampo/metabolismo , Ketamina/toxicidade , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Piroptose/fisiologia , Animais , Animais Recém-Nascidos , Inibidores de Caspase/farmacologia , Inibidores de Caspase/uso terapêutico , Disfunção Cognitiva/induzido quimicamente , Disfunção Cognitiva/prevenção & controle , Dipeptídeos/farmacologia , Dipeptídeos/uso terapêutico , Antagonistas de Aminoácidos Excitatórios/toxicidade , Feminino , Furanos/farmacologia , Furanos/uso terapêutico , Hipocampo/efeitos dos fármacos , Indenos/farmacologia , Indenos/uso terapêutico , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Aprendizagem em Labirinto/fisiologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/antagonistas & inibidores , Células PC12 , Piroptose/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Sulfonamidas/farmacologia , Sulfonamidas/uso terapêutico , para-Aminobenzoatos/farmacologia , para-Aminobenzoatos/uso terapêuticoRESUMO
Caspases are an evolutionary conserved family of cysteine-dependent proteases that are involved in many vital cellular processes including apoptosis, proliferation, differentiation and inflammatory response. Dysregulation of caspase-mediated apoptosis and inflammation has been linked to the pathogenesis of various diseases such as inflammatory diseases, neurological disorders, metabolic diseases, and cancer. Multiple caspase inhibitors have been designed and synthesized as a potential therapeutic tool for the treatment of cell death-related pathologies. However, only a few have progressed to clinical trials because of the consistent challenges faced amongst the different types of caspase inhibitors used for the treatment of the various pathologies, namely an inadequate efficacy, poor target specificity, or adverse side effects. Importantly, a large proportion of this failure lies in the lack of understanding various caspase functions. To overcome the current challenges, further studies on understanding caspase function in a disease model is a fundamental requirement to effectively develop their inhibitors as a treatment for the different pathologies. Therefore, the present review focuses on the descriptive properties and characteristics of caspase inhibitors known to date, and their therapeutic application in animal and clinical studies. In addition, a brief discussion on the achievements, and current challenges faced, are presented in support to providing more perspectives for further development of successful therapeutic caspase inhibitors for various diseases.
Assuntos
Inibidores de Caspase/uso terapêutico , Animais , Caspases/química , Caspases/metabolismo , Ensaios Clínicos como Assunto , Modelos Animais de Doenças , HumanosRESUMO
As a double-stranded DNA (dsDNA) sensor, the PYHIN family member absent in melanoma 2 (AIM2) is an essential component of the inflammasome families. Activation of AIM2 by dsDNA leads to the assembly of cytosolic multimolecular complexes termed the AIM2 inflammasome, resulting in activation of caspase-1, the maturation and secretion of pro-inflammatory cytokines interleukin (IL)-1ß and IL-18, and pyroptosis. Multiple central nervous system (CNS) diseases are accompanied by immune responses and inflammatory cascade. As the resident macrophage cells, microglia cells act as the first and main form of active immune defense in the CNS. AIM2 is highly expressed in microglia as well as astrocytes and neurons and is essential in neurodevelopment. In this review, we highlight the recent progress on the role of AIM2 inflammasome in CNS disorders, including cerebral stroke, brain injury, neuropsychiatric disease, neurodegenerative diseases, and glioblastoma.
Assuntos
Doenças do Sistema Nervoso Central/metabolismo , Proteínas de Ligação a DNA/metabolismo , Mediadores da Inflamação/antagonistas & inibidores , Mediadores da Inflamação/metabolismo , Animais , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Produtos Biológicos/farmacologia , Produtos Biológicos/uso terapêutico , Caspase 1/metabolismo , Inibidores de Caspase/farmacologia , Inibidores de Caspase/uso terapêutico , Doenças do Sistema Nervoso Central/tratamento farmacológico , Doenças do Sistema Nervoso Central/patologia , DNA/metabolismo , Humanos , Piroptose/efeitos dos fármacos , Piroptose/fisiologiaRESUMO
Alcoholic liver disease (ALD) is one of the pathogenic factors of chronic liver disease with the highest clinical morbidity worldwide. Ursolic acid (UA), a pentacyclic terpenoid carboxylic acid, has shown many health benefits including antioxidative, anti-inflammatory, anticancer, and hepatoprotective activities. We previously found that UA was metabolized in vivo into epoxy-modified UA containing an epoxy electrophilic group and had the potential to react with nucleophilic groups. In this study we prepared an alkynyl-modified UA (AM-UA) probe for tracing and capturing the target protein of UA from liver in mice, then investigated the mode by which UA bound to its target in vivo. By conducting proteome identification and bioinformatics analysis, we identified caspase-3 (CASP3) as the primary target protein of UA associated with liver protection. Molecule docking analysis showed that the epoxy group of the UA metabolite reacted with Cys-163 of CASP3, forming a covalent bond with CASP3. The binding mode of the UA metabolites (UA, CM-UA, and EM-UA) was verified by biochemical evaluation, demonstrating that the epoxy group produced by metabolism played an important role in the inhibition of CASP3. In alcohol-treated HepG2 cells, pretreatment with the UA metabolite (10 µM) irreversibly inhibited CASP3 activities, and subsequently decreased the cleavage of PARP and cell apoptosis. Finally, pre-administration of UA (20-80 mg· kg-1 per day, ig, for 1 week) dose-dependently alleviated alcohol-induced liver injury in mice mainly via the inhibition of CASP3. In conclusion, this study demonstrates that UA is a valuable lead compound for the treatment of ALD.
Assuntos
Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Inibidores de Caspase/uso terapêutico , Hepatopatias Alcoólicas/tratamento farmacológico , Fígado/efeitos dos fármacos , Triterpenos/uso terapêutico , Sequência de Aminoácidos , Animais , Caspase 3/química , Inibidores de Caspase/metabolismo , Cisteína/química , Compostos de Epóxi/química , Compostos de Epóxi/uso terapêutico , Células Hep G2 , Hepatócitos/efeitos dos fármacos , Humanos , Fígado/enzimologia , Fígado/patologia , Hepatopatias Alcoólicas/enzimologia , Hepatopatias Alcoólicas/patologia , Masculino , Camundongos , Simulação de Acoplamento Molecular , Poli(ADP-Ribose) Polimerases/metabolismo , Ligação Proteica , Alinhamento de Sequência , Triterpenos/metabolismo , Ácido UrsólicoRESUMO
Programmed cell death (PCD), including apoptosis, apoptotic necrosis, and pyroptosis, is involved in various organ dysfunction syndromes. Recent studies have revealed that a substrate of caspase-3, gasdermin E (GSDME), functions as an effector for pyroptosis; however, few inhibitors have been reported to prevent pyroptosis mediated by GSDME. Here, we developed a class of GSDME-derived inhibitors containing the core structure of DMPD or DMLD. Ac-DMPD-CMK and Ac-DMLD-CMK could directly bind to the catalytic domains of caspase-3 and specifically inhibit caspase-3 activity, exhibiting a lower IC50 than that of Z-DEVD-FMK. Functionally, Ac-DMPD/DMLD-CMK substantially inhibited both GSDME and PARP cleavage by caspase-3, preventing apoptotic and pyroptotic events in hepatocytes and macrophages. Furthermore, in a mouse model of bile duct ligation that mimics intrahepatic cholestasis-related acute hepatic failure, Ac-DMPD/DMLD-CMK significantly alleviated liver injury. Together, this study not only identified two specific inhibitors of caspase-3 for investigating PCD but also, more importantly, shed light on novel lead compounds for treating liver failure and organ dysfunctions caused by PCD.
Assuntos
Clorometilcetonas de Aminoácidos/uso terapêutico , Caspase 3/metabolismo , Inibidores de Caspase/uso terapêutico , Hepatopatias/prevenção & controle , Oligopeptídeos/uso terapêutico , Substâncias Protetoras/uso terapêutico , Clorometilcetonas de Aminoácidos/química , Animais , Apoptose/efeitos dos fármacos , Ductos Biliares/cirurgia , Inibidores de Caspase/química , Linhagem Celular Tumoral , Humanos , Ligadura , Masculino , Camundongos Endogâmicos C57BL , Simulação de Acoplamento Molecular , Oligopeptídeos/química , Fragmentos de Peptídeos/química , Substâncias Protetoras/química , Piroptose/efeitos dos fármacos , Receptores de Estrogênio/químicaRESUMO
Caspases are a family of cysteine proteases well known for their central roles during apoptosis and inflammation. They also intervene in non-apoptotic regulated cell death pathways and contribute to a large number of physiological mechanisms. The development of therapeutic approaches targeting caspases has generated strong industrial interest since the 1990s, prompting intense research on biological mechanisms, and the development of numerous synthetic inhibitors. Most of these inhibitors are derivatives of peptides or mimetics capable of interacting with the active site of caspases. However, the structural conservation between the different caspases is a challenge for the development of selective inhibitors. To date 5 caspase inhibitors, targeting either Caspase-1, -2 or multiple caspases, have been investigated in clinical settings, and there is still no marketing authorization. The Pan-caspase inhibitor emricasan reached clinical phase III and was proven to be safe but failed to demonstrate efficacy against NASH. Contrary to initial assumptions, selective Caspase-3 inhibitors have not reached the clinical level, while QPI-1007, a siRNA directed against Caspase-2, is currently undergoing a multicentric phase III clinical study for the treatment of ischemic optic neuropathies.
TITLE: Inhibition des caspases - De la biologie et thanatologie cellulaires au développement clinique de candidats médicaments. ABSTRACT: Les caspases sont une famille de cystéines protéases bien connues pour leurs rôles centraux au cours de l'apoptose et de l'inflammation. Elles interviennent aussi dans des voies de mort cellulaire régulées non-apoptotiques, et contribuent à de très nombreux mécanismes physiologiques. Le développement d'approches thérapeutiques ciblant les caspases a engendré un fort intérêt industriel dès les années 1990, suscitant d'intenses recherches sur les mécanismes biologiques, et conduisant à la mise au point de nombreux inhibiteurs synthétiques. La plupart de ces inhibiteurs sont des dérivés de peptides, ou mimétiques, capables d'interagir avec le site actif des caspases. Cependant, la conservation structurelle observée entre les différentes caspases est un défi pour le développement d'inhibiteurs sélectifs. À ce jour, cinq inhibiteurs de caspases ont été évalués pour leur efficacité clinique, mais aucune autorisation de mise sur le marché n'a été délivrée à ce jour. Contrairement aux présomptions initiales, les inhibiteurs sélectifs de la Caspase-3 n'ont pas atteint le stade d'essais cliniques, alors que le QPI-1007, un siARN dirigé contre la Caspase-2, a fait l'objet d'une étude clinique de phase III pour le traitement de neuropathies optiques ischémiques.
Assuntos
Inibidores de Caspase/uso terapêutico , Biologia Celular/tendências , Senescência Celular/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Desenvolvimento de Medicamentos/métodos , Desenvolvimento de Medicamentos/tendências , Humanos , Terapia de Alvo Molecular/métodos , Terapia de Alvo Molecular/tendências , Transdução de Sinais/efeitos dos fármacos , TanatologiaRESUMO
The paracaspase MALT1 has gained increasing interest as a target for the treatment of subsets of lymphomas as well as autoimmune diseases, and there is a need for suitable compounds to explore the therapeutic potential of this target. Here, we report the optimization of the in vivo potency of pyrazolopyrimidines, a class of highly selective allosteric MALT1 inhibitors. High doses of the initial lead compound led to tumor stasis in an activated B-cell-like (ABC) diffuse large B-cell lymphoma (DLBCL) xenograft model, but this compound suffered from a short in vivo half-life and suboptimal potency in whole blood. Guided by metabolism studies, we identified compounds with reduced metabolic clearance and increased in vivo half-life. In the second optimization step, masking one of the hydrogen-bond donors of the central urea moiety through an intramolecular interaction led to improved potency in whole blood. This was associated with improved in vivo potency in a mechanistic model of B cell activation. The optimized compound led to tumor regression in a CARD11 mutant ABC-DLBCL lymphoma xenograft model.
Assuntos
Sangue/metabolismo , Inibidores de Caspase/uso terapêutico , Proteína de Translocação 1 do Linfoma de Tecido Linfoide Associado à Mucosa/antagonistas & inibidores , Pirazóis/uso terapêutico , Pirimidinas/uso terapêutico , Ureia/uso terapêutico , Animais , Antineoplásicos/síntese química , Antineoplásicos/metabolismo , Antineoplásicos/farmacocinética , Antineoplásicos/uso terapêutico , Inibidores de Caspase/síntese química , Inibidores de Caspase/metabolismo , Inibidores de Caspase/farmacocinética , Linhagem Celular Tumoral , Feminino , Meia-Vida , Humanos , Camundongos Endogâmicos BALB C , Camundongos SCID , Microssomos Hepáticos/metabolismo , Neoplasias/tratamento farmacológico , Pirazóis/síntese química , Pirazóis/metabolismo , Pirazóis/farmacocinética , Pirimidinas/síntese química , Pirimidinas/metabolismo , Pirimidinas/farmacocinética , Ratos Sprague-Dawley , Ovinos , Ureia/síntese química , Ureia/metabolismo , Ureia/farmacocinética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
MALT1 plays a central role in immune cell activation by transducing NF-κB signaling, and its proteolytic activity represents a key node for therapeutic intervention. Two cycles of scaffold morphing of a high-throughput biochemical screening hit resulted in the discovery of MLT-231, which enabled the successful pharmacological validation of MALT1 allosteric inhibition in preclinical models of humoral immune responses and B-cell lymphomas. Herein, we report the structural activity relationships (SARs) and analysis of the physicochemical properties of a pyrazolopyrimidine-derived compound series. In human T-cells and B-cell lymphoma lines, MLT-231 potently and selectively inhibits the proteolytic activity of MALT1 in NF-κB-dependent assays. Both in vitro and in vivo profiling of MLT-231 support further optimization of this in vivo tool compound toward preclinical characterization.
Assuntos
Inibidores de Caspase/uso terapêutico , Proteína de Translocação 1 do Linfoma de Tecido Linfoide Associado à Mucosa/antagonistas & inibidores , Neoplasias/tratamento farmacológico , Ureia/análogos & derivados , Ureia/uso terapêutico , Animais , Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Inibidores de Caspase/síntese química , Inibidores de Caspase/farmacologia , Descoberta de Drogas , Feminino , Humanos , Imunidade Humoral/efeitos dos fármacos , Masculino , Camundongos Endogâmicos BALB C , Estrutura Molecular , Pirazóis/síntese química , Pirazóis/farmacologia , Pirazóis/uso terapêutico , Pirimidinas/síntese química , Pirimidinas/farmacologia , Pirimidinas/uso terapêutico , Ratos Sprague-Dawley , Relação Estrutura-Atividade , Linfócitos T/efeitos dos fármacos , Ureia/farmacologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
The cysteinyl aspartate protease (caspase, or CASP) gene family plays a significant role in programmed cell death, inflammation and immunity. However, the correlation between CASP family members and prognosis and tumor-infiltrating lymphocytes in different tumors has not been determined. We investigated the role of CASP genes in cancer prognosis and their relationship with clinicopathological parameters. We also evaluated the correlation between the expression of CASP family members and cancer immune infiltration and evaluated whether these molecules can be used as targets for immunotherapy. The CASP1/2/4/5/7/9 genes may represent prognostic factors and therapeutic targets for breast cancer, hepatocellular carcinoma and pancreatic cancer. Another finding is that the CASP1/4/5 genes help to regulate innate immunity and T cell immunity and may also have an important effect on tumor checkpoint inhibition. These findings may elucidate the roles played by CASP family members in cancer progression and identify strategies to promote collaborative activities in the context of immunotherapy.
Assuntos
Caspases/metabolismo , Neoplasias/enzimologia , Biomarcadores/metabolismo , Neoplasias da Mama/enzimologia , Neoplasias da Mama/genética , Neoplasias da Mama/imunologia , Neoplasias da Mama/patologia , Carcinoma Hepatocelular/enzimologia , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/imunologia , Carcinoma Hepatocelular/patologia , Inibidores de Caspase/uso terapêutico , Caspases/genética , Feminino , Humanos , Inibidores de Checkpoint Imunológico/uso terapêutico , Neoplasias Hepáticas/enzimologia , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/imunologia , Neoplasias Hepáticas/patologia , Linfócitos do Interstício Tumoral , Família Multigênica , Neoplasias/tratamento farmacológico , Neoplasias/imunologia , Neoplasias/mortalidade , Neoplasias Pancreáticas/enzimologia , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/imunologia , Neoplasias Pancreáticas/patologia , Prognóstico , Mapas de Interação de Proteínas , RNA Mensageiro/metabolismo , Resultado do TratamentoRESUMO
High-dose radiation activates caspases in tumor cells to produce abundant DNA fragments for DNA sensing in antigen-presenting cells, but the intrinsic DNA sensing in tumor cells after radiation is rather limited. Here we demonstrate that irradiated tumor cells hijack caspase 9 signaling to suppress intrinsic DNA sensing. Instead of apoptotic genomic DNA, tumor-derived mitochondrial DNA triggers intrinsic DNA sensing. Specifically, loss of mitochondrial DNA sensing in Casp9-/- tumors abolishes the enhanced therapeutic effect of radiation. We demonstrated that combining emricasan, a pan-caspase inhibitor, with radiation generates synergistic therapeutic effects. Moreover, loss of CASP9 signaling in tumor cells led to adaptive resistance by upregulating programmed death-ligand 1 (PD-L1) and resulted in tumor relapse. Additional anti-PD-L1 blockade can further overcome this acquired immune resistance. Therefore, combining radiation with a caspase inhibitor and anti-PD-L1 can effectively control tumors by sequentially blocking both intrinsic and extrinsic inhibitory signaling.
Assuntos
Antineoplásicos Imunológicos/uso terapêutico , Caspase 9/metabolismo , Inibidores de Caspase/uso terapêutico , Quimiorradioterapia/métodos , Neoplasias Colorretais/terapia , Ácidos Pentanoicos/uso terapêutico , Animais , Antígeno B7-H1/genética , Antígeno B7-H1/metabolismo , Caspase 9/genética , Linhagem Celular Tumoral , Modelos Animais de Doenças , Resistencia a Medicamentos Antineoplásicos/genética , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Transplante de Neoplasias , Transdução de Sinais , Regulação para CimaAssuntos
Carcinoma Hepatocelular/enzimologia , Inibidores de Caspase/uso terapêutico , Caspases/metabolismo , Neoplasias Hepáticas/enzimologia , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Apoptose/efeitos dos fármacos , Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/patologia , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/antagonistas & inibidores , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteínas com Domínio LIM/antagonistas & inibidores , Proteínas com Domínio LIM/metabolismo , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/patologia , Proteínas Musculares/antagonistas & inibidores , Proteínas Musculares/metabolismoRESUMO
Liver cell death has an essential role in nonalcoholic steatohepatitis (NASH). The activity of the energy sensor adenosine monophosphate (AMP)-activated protein kinase (AMPK) is repressed in NASH. Liver-specific AMPK knockout aggravated liver damage in mouse NASH models. AMPK phosphorylated proapoptotic caspase-6 protein to inhibit its activation, keeping hepatocyte apoptosis in check. Suppression of AMPK activity relieved this inhibition, rendering caspase-6 activated in human and mouse NASH. AMPK activation or caspase-6 inhibition, even after the onset of NASH, improved liver damage and fibrosis. Once phosphorylation was decreased, caspase-6 was activated by caspase-3 or -7. Active caspase-6 cleaved Bid to induce cytochrome c release, generating a feedforward loop that leads to hepatocyte death. Thus, the AMPK-caspase-6 axis regulates liver damage in NASH, implicating AMPK and caspase-6 as therapeutic targets.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Caspase 6/metabolismo , Fígado/enzimologia , Fígado/patologia , Hepatopatia Gordurosa não Alcoólica/enzimologia , Hepatopatia Gordurosa não Alcoólica/patologia , Proteínas Quinases Ativadas por AMP/genética , Animais , Apoptose/genética , Caspase 3/metabolismo , Caspase 7/metabolismo , Inibidores de Caspase/farmacologia , Inibidores de Caspase/uso terapêutico , Ativação Enzimática , Hepatócitos/enzimologia , Hepatócitos/patologia , Humanos , Camundongos , Camundongos Knockout , Hepatopatia Gordurosa não Alcoólica/tratamento farmacológico , FosforilaçãoRESUMO
Viral myocarditis caused by Coxsackievirus B (CVB) infection is a severe inflammatory disease of the myocardium, which may develop to cardiomyopathy and heart failure. No effective specific treatment is available. Our previous study demonstrated that suppression of proinflammatory caspase-1 activation effectively inhibited CVB replication. N-acetyl cysteine (NAC) is a widely used antioxidant. In this study, we found that NAC significantly alleviated the myocardial injury caused by CVB type 3 (CVB3) under in vivo condition. Importantly, NAC treatment simultaneously suppressed viral replication and inflammatory response in both myocardium and cell culture. The antiviral and anti-inflammation mechanism of NAC, while independent of its antioxidant property, relies on its inhibition on caspase-1 activation. Moreover, NAC promotes procaspase-1 degradation via ubiquitin proteasome system, which further contributes to caspase-1 down-regulation. NAC also inhibits the activity of viral proteases. Taken together, this study shows that NAC exerts potent anti-CVB and anti-inflammation effect through targeting caspase-1. Given that NAC is a clinically approved medicine, we recommend NAC as a valuable therapeutic agent for viral myocarditis caused by CVB.
Assuntos
Acetilcisteína/uso terapêutico , Antivirais/uso terapêutico , Infecções por Coxsackievirus/tratamento farmacológico , Miocardite/tratamento farmacológico , Replicação Viral/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Inibidores de Caspase/uso terapêutico , Infecções por Coxsackievirus/complicações , Enterovirus Humano B/efeitos dos fármacos , Enterovirus Humano B/fisiologia , Células HeLa , Humanos , Inflamação/tratamento farmacológico , Inflamação/virologia , Camundongos Endogâmicos BALB C , Miocardite/virologia , Complexo de Endopeptidases do Proteassoma/metabolismo , Organismos Livres de Patógenos EspecíficosRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: The fruits of Nandina domestica Thunb. have served as folk medicines in Chinese and Japanese tradition for treatment of several tumors including pharynx tumor and tooth abscess for many years, yet its exact mechanism of action is not yet known. AIM OF THE STUDY: The study targets the identification of the main constituents of the fruits extracts and investigation of their mode of action in cancer therapy via pharmacology-based analysis and molecular docking. MATERIALS AND METHODS: The different extracts of N. domestica Thunb. were analyzed via UPLC-MS/MS for identification of their active constituents. STITCH, DAVID, KEGG and STRING database were utilized for construction of compound-target and compound-target-pathway networks using Cytoscape 3.2.1. Molecular docking analysis of the top hit compounds was performed against the identified top hit molecular targets in the constructed networks. In vitro-testing of Nandina domestica Thunb. against colorectal cancer cell lines was carried out and correlated to the chemical profile of the extract to identify important biomarkers. The ADME properties of the active compounds were also evaluated. RESULTS: 22 compounds were identified by UPLC-MS/MS analysis and were forwarded to network pharmacology-based analysis. Results showed the enrichment of 5 compounds and 4 molecular targets in the network namely; AKT1, CASP3, MAPK1 and TP53. The pathway analysis of the identified targets revealed that 15 cancer-related pathways were enriched including colorectal cancer, endometrial cancer and small-cell lung cancer. In-vitro testing of the extracts against colo-rectal cancer cell lines revealed the fractions enriched in the identified hit compounds were indeed the most active as revealed from the HCA-heat-map. ADME results showed that all compounds were drug-like candidates showing acceptable values according to Lipinski's rule. CONCLUSIONS: Network pharmacology analysis revealed that the compounds isoquercitrin, quercitrin, berberine, chlorogenic acid and caffeic acid showed strong synergistic interactions with the cancer-related targets and pathways. It could be concluded that N. domestica Thunb. constituents affect both apoptosis and Akt-signaling pathways during the stages of early and intermediate adenoma through interaction with the targets CASP3 and MAPK1 (ErC2) while during the stages of late adenoma and carcinoma, the compounds acts through the p53 and ErbB signaling pathways.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Neoplasias/tratamento farmacológico , Ranunculales/química , Apoptose/efeitos dos fármacos , Apoptose/genética , Caspase 3/metabolismo , Inibidores de Caspase/química , Inibidores de Caspase/farmacologia , Inibidores de Caspase/uso terapêutico , Linhagem Celular Tumoral , Ensaios de Seleção de Medicamentos Antitumorais , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/uso terapêutico , Frutas/química , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Medicina Tradicional Chinesa/métodos , Proteína Quinase 1 Ativada por Mitógeno/antagonistas & inibidores , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Simulação de Acoplamento Molecular , Neoplasias/genética , Neoplasias/patologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Espectrometria de Massas em TandemRESUMO
Stroke is a life-threatening neurological disease with limited therapeutic options. Inflammation is believed to be involved in the pathogenesis of ischemic stroke and contribute to the degree of brain injury. Vx-765 is a potent, selective, small-molecule caspase-1 inhibitor. Current studies have shown the anti-inflammatory properties of vx-765 in various disease; however, the impact of vx-765 on the ischemic stroke is still unclear. In the present study, we determine the neuroprotective effect of vx-765 in mice subjected to transient middle cerebral artery occlusion (MCAO). We found that caspase-1 inhibition by administration of vx-765 ameliorated cerebral injury in mice after ischemic stroke by reducing infarct volume and ameliorating the neurological deficits. Mechanistically, we showed that the contribution of vx-765 to ischemic injuries may be associated with reducing microglial activation, and downregulating the production of associated pro inflammatory cytokines including IL-1ß, TNF-α, and iNOS, as well as upregulating anti-inflammatory cytokines such as TGF-ß and YM-1. Additionally, vx-765 altered the phenotype of microglia via switching the microglia polarization toward M2 phenotype, as demonstrably related to inhibition of the NF-κB activation. Our findings indicate that vx-765 protects against MCAO injury and attenuated microglia mediated neuroinflammation primarily by shifting microglia polarization from M1 phenotype toward M2 phenotype. Vx-765 might be a potential therapeutic drug for ameliorating ischemic stroke.
Assuntos
Inibidores de Caspase/uso terapêutico , Dipeptídeos/uso terapêutico , NF-kappa B/antagonistas & inibidores , Fármacos Neuroprotetores/uso terapêutico , Acidente Vascular Cerebral/tratamento farmacológico , para-Aminobenzoatos/uso terapêutico , Animais , Caspase 1/imunologia , Modelos Animais de Doenças , Infarto da Artéria Cerebral Média/tratamento farmacológico , Infarto da Artéria Cerebral Média/imunologia , Infarto da Artéria Cerebral Média/patologia , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/patologia , Masculino , Camundongos Endogâmicos C57BL , Microglia/efeitos dos fármacos , Microglia/imunologia , Microglia/patologia , NF-kappa B/imunologia , Neuroproteção/efeitos dos fármacos , Acidente Vascular Cerebral/imunologia , Acidente Vascular Cerebral/patologiaRESUMO
Male infertility is due to genetics, hormonal or environmental causes, or is idiopathic. Azoospermia is linked to local testicular microenvironment deregulation, with inflammatory cells present in the 15% of testicular biopsies of infertile patients. As widely reported, spermatogenesis and steroidogenesis are controlled by local immunoregulatory agents produced by immune and nonimmune cells. Moreover IL-6R, TNFR1, Fas and IL-1R are expressed on germ cells, indicating a direct action of pro-inflammatory agents on these cells. Beyond the known function of cytokines and nitric oxide on testicular function at the stable levels present in the normal testis, this review focalises on the effect of pro-inflammatory factors on germ cell survival and death when inflammatory conditions are established in the testis. As no cure for male infertility has been found up to the present, intracytoplasmic sperm injection is the therapeutic option for azoospermic patients who wish to achieve genetic parenthood. Therapies with antioxidant and anti-inflammatory agents in experimental models of testicular damage have been successful. However, clinical implementation is uncertain in cases with a prolonged inflammatory state of the testis. Therapies offering multiple approaches to treat infertility by restoring the spermatogonial stem cell niche and protecting germ cells from apoptosis should be considered.
Assuntos
Apoptose/imunologia , Azoospermia/imunologia , Orquite/imunologia , Espermatogônias/patologia , Testículo/patologia , Células-Tronco Germinativas Adultas/efeitos dos fármacos , Células-Tronco Germinativas Adultas/fisiologia , Animais , Apoptose/efeitos dos fármacos , Azoospermia/tratamento farmacológico , Azoospermia/patologia , Biópsia , Inibidores de Caspase/farmacologia , Inibidores de Caspase/uso terapêutico , Modelos Animais de Doenças , Humanos , Masculino , Orquite/complicações , Orquite/patologia , Espermatogênese/efeitos dos fármacos , Espermatogênese/imunologia , Espermatogônias/imunologia , Testículo/citologia , Testículo/imunologia , Agentes Urológicos/farmacologia , Agentes Urológicos/uso terapêuticoRESUMO
As a main active ingredient of Fenugreek, trigonelline has protective efficiency on type 2 diabetes and diabetic peripheral neuropathy in rats. This study investigates the protection of trigonelline on hyperglycemia, ß cell apoptosis, and inflammation in type 1 diabetic mice. Streptozotocin (160â¯mg/kg) was intraperitoneal injected to induce diabetic mice. There were 4 groups: normal control, diabetes, trigonelline-treated diabetes, and insulin-treated diabetes. After 4-week treatment, levels of blood glucose, serum insulin, and inflammatory factors, ß cell apoptosis, insulin content, and oxidative stress parameters in pancreas were calculated. Pancreas was examined by immunohistochemistry staining and hematoxylin/eosin. Trigonelline significantly declined the levels of blood glucose, serum tumor necrosis factor-α, interleukin-6, and interleukin-1ß, while increased the levels of serum insulin and adiponectin in diabetic mice. Insulin content, glutathione concentration, serum activities of superoxide dismutase and catalase in pancreas, and pancreas to body weight ratio were remarkably decreased, while serum malondialdehyde concentration was increased in diabetic mice. Trigonelline treatment restored the above mentioned parameters. Trigonelline even suppresses ß cell apoptosis via downregulating caspase 3 expression. These results imply that trigonelline protects diabetic mice mediated by decreasing blood glucose, increasing insulin expression in ß cells, regulating inflammatory response, suppressing ß cells apoptosis partly by downregulating caspase 3 expression, and raising antioxidant enzyme activity.
Assuntos
Alcaloides/farmacologia , Caspase 3/metabolismo , Inibidores de Caspase/farmacologia , Diabetes Mellitus Experimental/patologia , Diabetes Mellitus Tipo 1/patologia , Células Secretoras de Insulina/efeitos dos fármacos , Células Secretoras de Insulina/patologia , Animais , Glicemia/metabolismo , Peso Corporal/efeitos dos fármacos , Inibidores de Caspase/uso terapêutico , Citoproteção/efeitos dos fármacos , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/enzimologia , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 1/tratamento farmacológico , Diabetes Mellitus Tipo 1/enzimologia , Diabetes Mellitus Tipo 1/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Insulina/biossíntese , Insulina/sangue , Insulina/metabolismo , Células Secretoras de Insulina/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Respiratory diseases and asthma, in particular, are nowadays a global health problem. In Rwanda, some traditional healers claim to treat asthma with plant-based recipes, though there is no scientific proof so far. AIM OF THE STUDY: Our study aimed at evaluating the toxicity and the anti-inflammatory effect of plant recipes used in Rwanda against asthma in order to select potential candidates for further characterization of the active compounds. MATERIALS AND METHODS: Water (aqueous) and methanol-dichloromethane (organic) extracts from selected folkloric recipes were submitted for toxicity test on THP-1 derived macrophages using CellTiter-Glo Luminescent Cell Viability Assay. The evaluation of the anti-inflammatory effect of the plant extracts was carried out using the Caspase-Glo 1 Inflammasome assay on THP-1 -derived macrophages. RESULTS: Most of both organic and aqueous extract showed more than 95% of cell viability up to 200⯵g/ml, except for R03Cn organic extract that inhibited 25% of the cell viability. Plant extracts inhibited caspase-1 activation in THP-1 derived macrophages in a dose-dependent manner. Four extracts (R03Cn and R07Kn aqueous extracts, R10MK and R19Sz organic extracts) strongly downregulated the activation of caspase-1 (more than 70% at 50⯵g/ml). In general, organic extracts exhibited better caspase-1 inhibitory effects than their aqueous counterparts. CONCLUSIONS: The inhibition of inflammasome/caspase-1 is one of key mechanisms of action in asthma. Some traditional recipes are active on this mechanism and are thus strong candidates for the treatment of asthma and other inflammasome-mediated diseases. Further investigations are needed to characterize active molecules.