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1.
Int J Mol Sci ; 22(24)2021 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-34948191

RESUMO

Apoptosis signal-regulating kinase (ASK) 1, a member of the mitogen-activated protein kinase kinase kinase (MAP3K) family, modulates diverse responses to oxidative and endoplasmic reticulum (ER) stress and calcium influx. As a crucial cellular stress sensor, ASK1 activates c-Jun N-terminal kinases (JNKs) and p38 MAPKs. Their excessive and sustained activation leads to cell death, inflammation and fibrosis in various tissues and is implicated in the development of many neurological disorders, such as Alzheimer's, Parkinson's and Huntington disease and amyotrophic lateral sclerosis, in addition to cardiovascular diseases, diabetes and cancer. However, currently available inhibitors of JNK and p38 kinases either lack efficacy or have undesirable side effects. Therefore, targeted inhibition of their upstream activator, ASK1, stands out as a promising therapeutic strategy for treating such severe pathological conditions. This review summarizes recent structural findings on ASK1 regulation and its role in various diseases, highlighting prospects for ASK1 inhibition in the treatment of these pathologies.


Assuntos
MAP Quinase Quinase Quinase 5/metabolismo , MAP Quinase Quinase Quinase 5/fisiologia , Proteínas 14-3-3/metabolismo , Animais , Apoptose/fisiologia , Proteínas Reguladoras de Apoptose/metabolismo , Estresse do Retículo Endoplasmático , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , MAP Quinase Quinase Quinase 5/genética , MAP Quinase Quinase Quinase 5/ultraestrutura , MAP Quinase Quinase Quinases/genética , MAP Quinase Quinase Quinases/metabolismo , Sistema de Sinalização das MAP Quinases , Oxirredução , Estresse Oxidativo , Fosforilação , Mapas de Interação de Proteínas/genética , Mapas de Interação de Proteínas/fisiologia , Transdução de Sinais/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
2.
Int J Mol Sci ; 22(18)2021 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-34576095

RESUMO

Titanium dioxide nanoparticles (TiO2NPs) are widely used in industrial and medicinal fields and in various consumer products, and their increasing use has led to an increase in the number of toxicity studies; however, studies investigating the underlying toxicity mechanism have been rare. In this study, we evaluated potential toxic effects of TiO2NPs exposure on lungs as well as the development of asthma through the ovalbumin (OVA)-induced mouse model of asthma. Furthermore, we also investigated the associated toxic mechanism. TiO2NPs caused pulmonary toxicity by exacerbating the inflammatory response, indicated by an increase in the number and level of inflammatory cells and mediators, respectively. OVA-induced asthma exposed mice to TiO2NPs led to significant increases in inflammatory mediators, cytokines, and airway hyperresponsiveness compared with those in non-exposed asthmatic mice. This was also accompanied by increased inflammatory cell infiltration and mucus production in the lung tissues. Additionally, TiO2NPs decreased the expression of B-cell lymphoma 2 (Bcl2) and the expressions of thioredoxin-interacting protein (TXNIP), phospho-apoptosis signal-regulating kinase 1, Bcl2-associated X, and cleaved-caspase 3 were escalated in the lungs of asthmatic mice compared with those in non-exposed asthmatic mice. These responses were consistent with in vitro results obtained using human airway epithelial cells. TiO2NPs treated cells exhibited an increase in the mRNA and protein expression of interleukin (IL)-1ß, IL-6, and tumor necrosis factor-α with an elevation of TXNIP signaling compared to non-treated cells. Moreover, pathophysiological changes induced by TiO2NP treatment were significantly decreased by TXNIP knockdown in airway epithelial cells. Overall, TiO2NP exposure induced toxicological changes in the respiratory tract and exacerbated the development of asthma via activation of the TXNIP-apoptosis pathway. These results provide insights into the underlying mechanism of TiO2NP-mediated respiratory toxicity.


Assuntos
Asma/patologia , Proteínas de Transporte/genética , Hipersensibilidade/patologia , Inflamação/patologia , Pulmão/patologia , Nanopartículas/toxicidade , Tiorredoxinas/genética , Titânio/toxicidade , Regulação para Cima/genética , Animais , Apoptose , Asma/sangue , Asma/complicações , Asma/genética , Líquido da Lavagem Broncoalveolar , Proteínas de Transporte/metabolismo , Caspase 3/metabolismo , Contagem de Células , Linhagem Celular , Fenômenos Químicos , Citocinas/biossíntese , Citocinas/genética , Citocinas/metabolismo , Modelos Animais de Doenças , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Hipersensibilidade/sangue , Hipersensibilidade/complicações , Hipersensibilidade/genética , Imunoglobulina E/sangue , Inflamação/sangue , Inflamação/genética , Mediadores da Inflamação/metabolismo , MAP Quinase Quinase Quinase 5/metabolismo , Camundongos , Muco/metabolismo , Nanopartículas/ultraestrutura , Ovalbumina , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Hipersensibilidade Respiratória/complicações , Tiorredoxinas/metabolismo , Regulação para Cima/efeitos dos fármacos , Proteína X Associada a bcl-2/metabolismo
3.
Biomed Res Int ; 2021: 9884297, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34307672

RESUMO

We performed in vitro and in vivo experiments to explore the role of protein kinase C-binding protein 1 (PICK1), an intracellular transporter involved in oxidative stress-related neuronal diseases, in sepsis-related acute kidney injury (AKI). Firstly, PCR, western blotting, and immunohistochemistry were used to observe the expression of PICK1 after lipopolysaccharide- (LPS-) induced AKI. Secondly, by inhibiting PICK1 in vivo and silencing PICK1 in vitro, we further explored the effect of PICK1 on AKI. Finally, the relationship between PICK1 and oxidative stress and the related mechanisms were explored. We found that the expression of PICK1 was increased in LPS-induced AKI models both in vitro and in vivo. PICK1 silencing significantly aggravated LPS-induced apoptosis, accompanied by ROS production in renal tubular epithelial cells. FSC231, a PICK1-specific inhibitor, aggravated LPS-induced kidney injury. Besides, NAC (N-acetylcysteine), a potent ROS scavenger, significantly inhibited the PICK1-silencing-induced apoptosis. In conclusion, PICK1 might protect renal tubular epithelial cells from LPS-induced apoptosis by reducing excessive ROS, making PICK1 a promising preventive target in LPS-induced AKI.


Assuntos
Injúria Renal Aguda/etiologia , Injúria Renal Aguda/metabolismo , Proteínas de Transporte/metabolismo , Proteínas de Ciclo Celular/deficiência , Proteínas Nucleares/metabolismo , Sepse/complicações , Acetilcisteína/farmacologia , Injúria Renal Aguda/patologia , Animais , Apoptose , Proteínas de Ciclo Celular/antagonistas & inibidores , Proteínas de Ciclo Celular/metabolismo , Linhagem Celular , Modelos Animais de Doenças , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Humanos , Túbulos Renais Proximais/patologia , Lipopolissacarídeos , MAP Quinase Quinase Quinase 5/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Peróxidos/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
4.
J Pharmacol Sci ; 146(3): 136-148, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34030796

RESUMO

Despite the documented renoprotective effect of pentoxifylline (PTX), a non-selective phosphodiesterase-4 inhibitor, the studies appraised only its anti-inflammatory/-oxidant/-apoptotic capacities without assessment of the possible involved trajectories. Here, we evaluated the potential role of galectin-3 and the ASK-1/NF-κB p65 signaling pathway with its upstream/downstream signals in an attempt to unveil part of the cascades involved in the renotherapeutic effect using a renal bilateral ischemia/reperfusion (I/R) model. Rats were randomized into sham-operated, renal I/R (45 min/72 h) and I/R + PTX (100 mg/kg; p.o). Post-treatment with PTX improved renal function and abated serum levels of cystatin C, creatinine, BUN and renal KIM-1 content, effects that were reflected on an improvement of the I/R-induced renal histological changes. On the molecular level, PTX reduced renal contents of galectin-3, ASK-1 with its downstream molecule JNK and ERK1/2, as well as NF-κB p65 and HMGB1. This inhibitory effect extended also to suppress neutrophil infiltration, evidenced by diminishing ICAM-1 and MPO, as well as inflammatory cytokines (TNF-α/IL-18), oxidative stress (MDA/TAC), and caspase-3. The PTX novel renotherapeutic effect involved in part the inhibition of galectin-3 and ASK-1/JNK and ERK1/2/NF-κB/HMGB-1 trajectories to mitigate renal I/R injury and to provide basis for its anti-inflammatory, antioxidant, and anti-apoptotic impacts.


Assuntos
Galectina 3/metabolismo , Proteína HMGB1/metabolismo , Rim/irrigação sanguínea , MAP Quinase Quinase Quinase 5/metabolismo , Sistema de Sinalização das MAP Quinases , Pentoxifilina/farmacologia , Pentoxifilina/uso terapêutico , Inibidores de Fosfodiesterase/farmacologia , Inibidores de Fosfodiesterase/uso terapêutico , Traumatismo por Reperfusão/tratamento farmacológico , Traumatismo por Reperfusão/genética , Transdução de Sinais/genética , Transdução de Sinais/fisiologia , Fator de Transcrição RelA/metabolismo , Animais , Anti-Inflamatórios , Antioxidantes , Modelos Animais de Doenças , Masculino , Ratos Wistar
5.
Sci Rep ; 11(1): 9832, 2021 05 10.
Artigo em Inglês | MEDLINE | ID: mdl-33972601

RESUMO

We recently found that tumor necrosis factor-α (TNF-α) may be involved in neuronal cell death induced by methylmercury in the mouse brain. Here, we examined the cells involved in the induction of TNF-α expression by methylmercury in the mouse brain by in situ hybridization. TNF-α-expressing cells were found throughout the brain and were identified as microglia by immunostaining for ionized calcium binding adaptor molecule 1 (Iba1). Methylmercury induced TNF-α expression in mouse primary microglia and mouse microglial cell line BV2. Knockdown of apoptosis signal-regulating kinase 1 (ASK1), an inflammatory cytokine up-regulator that is responsible for reactive oxygen species (ROS), decreased methylmercury-induced TNF-α expression through decreased phosphorylation of p38 MAP kinase in BV2 cells. Suppression of methylmercury-induced reactive oxygen species (ROS) by antioxidant treatment largely abolished the induction of TNF-α expression and phosphorylation of p38 by methylmercury in BV2 cells. Finally, in mouse brain slices, the TNF-α antagonist (WP9QY) inhibited neuronal cell death induced by methylmercury, as did the p38 inhibitor SB203580 and liposomal clodronate (a microglia-depleting agent). These results indicate that methylmercury induces mitochondrial ROS that are involved in activation of the ASK1/p38 pathway in microglia and that this is associated with induction of TNF-α expression and neuronal cell death.


Assuntos
Encéfalo/patologia , Intoxicação do Sistema Nervoso por Mercúrio/patologia , Microglia/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Encéfalo/citologia , Linhagem Celular , Ácido Clodrônico/farmacologia , Modelos Animais de Doenças , Poluentes Ambientais/administração & dosagem , Poluentes Ambientais/toxicidade , Técnicas de Silenciamento de Genes , Humanos , Imidazóis/farmacologia , MAP Quinase Quinase Quinase 5/genética , MAP Quinase Quinase Quinase 5/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/genética , Masculino , Intoxicação do Sistema Nervoso por Mercúrio/etiologia , Compostos de Metilmercúrio/administração & dosagem , Compostos de Metilmercúrio/toxicidade , Camundongos , Microglia/patologia , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Neurônios/patologia , Peptídeos Cíclicos/farmacologia , Fosforilação/efeitos dos fármacos , Cultura Primária de Células , Piridinas/farmacologia , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
6.
Cell Death Dis ; 12(5): 425, 2021 04 30.
Artigo em Inglês | MEDLINE | ID: mdl-33931585

RESUMO

Serum deprivation-response protein (SDPR), a phosphatidylserine-binding protein, which is known to have a promising role in caveolar biogenesis and morphology. However, its function in hepatocellular carcinoma (HCC) was still largely unknown. In this study, we discussed the characterization and identification of SDPR, and to present it as a novel apoptosis candidate in the incidence of HCC. We identified 81 HCC cases with lower SDPR expression in the tumor tissues with the help of qRT-PCR assay, and lower SDPR expression was potentially associated with poor prognostication. The phenotypic assays revealed that cell proliferation, invasion, and migration were profoundly connected with SDPR, both in vivo and in vitro. The data obtained from the gene set enrichment analysis (GSEA) carried out on the liver hepatocellular carcinoma (LIHC), and also The Cancer Genome Atlas (TCGA) findings indicated that SDPR was involved in apoptosis and flow cytometry experiments further confirmed this. Furthermore, we identified the interaction between SDPR and apoptosis signal-regulating kinase 1 (ASK1), which facilitated the ASK1 N-terminus-mediated dimerization and increased ASK1-mediated signaling, thereby activating the JNK/p38 mitogen-activated protein kinases (MAPKs) and finally enhanced cell apoptosis. Overall, this work identified SDPR as a tumor suppressor, because it promoted apoptosis by activating ASK1-JNK/p38 MAPK pathways in HCC.


Assuntos
Carcinoma Hepatocelular/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Neoplasias Hepáticas/metabolismo , MAP Quinase Quinase Quinase 5/metabolismo , Sistema de Sinalização das MAP Quinases , Proteínas de Ligação a Fosfato/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Apoptose/fisiologia , Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Células Hep G2 , Xenoenxertos , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Pessoa de Meia-Idade , Transfecção
7.
Sci Rep ; 11(1): 10350, 2021 05 14.
Artigo em Inglês | MEDLINE | ID: mdl-33990641

RESUMO

trans-Fatty acids (TFAs) are food-derived fatty acids associated with various diseases including cardiovascular diseases. However, the underlying etiology is poorly understood. Here, we show a pro-apoptotic mechanism of TFAs such as elaidic acid (EA), in response to DNA interstrand crosslinks (ICLs) induced by cisplatin (CDDP). We previously reported that TFAs promote apoptosis induced by doxorubicin (Dox), a double strand break (DSB)-inducing agent, via a non-canonical apoptotic pathway independent of tumor suppressor p53 and apoptosis signal-regulating kinase (ASK1), a reactive oxygen species (ROS)-responsive kinase. However, here we found that in the case of CDDP-induced apoptosis, EA-mediated pro-apoptotic action was reversed by knockout of either p53 or ASK1, despite no increase in p53 apoptotic activity. Upon CDDP treatment, EA predominantly enhanced ROS generation, ASK1-p38/c-Jun N-terminal kinase (JNK) mitogen-activated protein kinase (MAPK) pathway activation, and ultimately cell death, all of which were suppressed either by co-treatment of the NADPH oxidase (Nox) inhibitor Apocynin, or by knocking out its regulatory protein, receptor-interacting protein 1 (RIP1). These results demonstrate that in response to CDDP ICLs, TFAs promote p53-dependent apoptosis through the enhancement of the Nox-RIP1-ASK1-MAPK pathway activation, providing insight into the diverse pathogenetic mechanisms of TFAs according to the types of DNA damage.


Assuntos
Dano ao DNA/efeitos dos fármacos , Gorduras Insaturadas na Dieta/toxicidade , Ácidos Oleicos/toxicidade , Acetofenonas/farmacologia , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Doenças Cardiovasculares/etiologia , Doenças Cardiovasculares/prevenção & controle , Cisplatino/farmacologia , Gorduras Insaturadas na Dieta/efeitos adversos , Células HEK293 , Humanos , MAP Quinase Quinase Quinase 5/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/genética , Camundongos , NADPH Oxidases/antagonistas & inibidores , NADPH Oxidases/metabolismo , Ácidos Oleicos/efeitos adversos , Oxirredução , Células RAW 264.7 , Espécies Reativas de Oxigênio/metabolismo , Proteína Supressora de Tumor p53/metabolismo
8.
Cell Death Dis ; 12(5): 451, 2021 05 06.
Artigo em Inglês | MEDLINE | ID: mdl-33958583

RESUMO

Metastasis-associated protein 2 (MTA2) is a transcription factor that is highly associated with matrix metalloproteinase 12 (MMP12). Thus, we hypothesized that MTA2 may regulate MMP12 expression and is involved in cervical cancer metastasis. Results showed that MTA2 and MMP12 were highly expressed in cervical cancer cells, and MTA2 knockdown reduced MMP12 expression and inhibited the metastasis of cervical cancer cells in xenograft mice. MMP12 knockdown did not influence the viability of cervical cancer cells but clearly inhibited cell migration and invasion both in vitro and in vivo. MMP12 was highly expressed in cervical tumor tissues and correlated with the poor survival rate of patients with cervical cancer. Further investigations revealed that p38 mitogen-activated protein kinase (p38), mitogen-activated protein kinase kinase 3 (MEK3), and apoptosis signal-regulating kinase 1 (ASK1) were involved in MMP12 downregulation in response to MTA2 knockdown. Results also demonstrated that p38-mediated Y-box binding protein1 (YB1) phosphorylation disrupted the binding of AP1 (c-Fos/c-Jun) to the MMP12 promoter, thereby inhibiting MMP12 expression and the metastatic potential of cervical cancer cells. Collectively, targeting both MTA2 and MMP12 may be a promising strategy for the treatment of cervical cancer.


Assuntos
Histona Desacetilases/metabolismo , MAP Quinase Quinase Quinase 5/metabolismo , Sistema de Sinalização das MAP Quinases , Metaloproteinase 12 da Matriz/biossíntese , Proteínas Repressoras/metabolismo , Fator de Transcrição AP-1/metabolismo , Neoplasias do Colo do Útero/metabolismo , Proteína 1 de Ligação a Y-Box/metabolismo , Animais , Feminino , Células HeLa , Xenoenxertos , Histona Desacetilases/genética , Humanos , MAP Quinase Quinase 3/metabolismo , Metaloproteinase 12 da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Oncogenes , Proteínas Repressoras/genética , Transfecção , Neoplasias do Colo do Útero/genética , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
9.
Med Sci Monit ; 27: e930429, 2021 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-33811209

RESUMO

BACKGROUND Acute lung injury (ALI) results from damage to the alveolar capillary endothelial cells and can result in acute respiratory distress syndrome (ARDS). This study aimed to investigate murine lung vascular endothelial cells (MLECs) damage in a murine model of lipopolysaccharide (LPS)-induced ALI. MATERIAL AND METHODS Mice were injected with LPS to induce an acute lung injury model. An adenovirus transfection system was used to overexpress or knockdown DUSP12 in mice. MLECs were isolated, cultured and transfected with DUSP12-overexpressing adenovirus or with DUSP12 siRNA to knockdown DUSP12. LPS was used to establish a cell injury model. ELISA and RT-PCR were used to examine cell inflammation. LPS-induced oxidative stress was also evaluated using commercial kits. RESULTS A decreased level of DUSP12 was observed in MLECs treated with LPS. DUSP12 overexpression in mice attenuated LPS-induced lung inflammation and lung injury, as reflected by reduced levels of proinflammatory cytokines. Mice with DUSP12 knockdown exhibited worsened lung inflammation and injury. In vitro, DUSP12 overexpression in endothelial cells ameliorated LPS-induced inflammation, apoptosis, and oxidative stress. DUSP12 silencing in endothelial cells aggravated LPS-induced inflammation, apoptosis, and oxidative stress. Furthermore, we found that DUSP12 directly bound to apoptosis signal-regulating kinase 1 (ASK1) to inhibit Jun N-terminal kinase activation (JNK). A JNK1/2 inhibitor and ASK1 siRNA ameliorated the exacerbating effects of DUSP12 knockdown in vitro. CONCLUSIONS Our data demonstrated that DUSP12 suppressed MLEC injury in response to LPS insult by regulating the ASK1/JNK pathway.


Assuntos
Lesão Pulmonar Aguda/metabolismo , Fosfatases de Especificidade Dupla/metabolismo , Células Endoteliais/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , MAP Quinase Quinase 4/metabolismo , MAP Quinase Quinase Quinase 5/metabolismo , Lesão Pulmonar Aguda/patologia , Animais , Células Cultivadas , Modelos Animais de Doenças , Fosfatases de Especificidade Dupla/genética , Células Endoteliais/patologia , Humanos , Lipopolissacarídeos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Transdução de Sinais
10.
Eur J Med Chem ; 220: 113482, 2021 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-33906048

RESUMO

Apoptosis signal-regulating kinase 1 (ASK1, MAP3K5), a member of the mitogen-activated protein kinase (MAPK) signaling pathway, is involved in cell survival, differentiation, stress response, and apoptosis. ASK1 kinase inhibition has emerged as a promising therapeutic strategy for inflammatory disease. A series of novel ASK1 inhibitors with 1H-indazole scaffold were designed, synthesized and evaluated for their ASK1 kinase activity and AP1-HEK293 cell inhibitory effect. Systematic structure-activity relationship (SAR) efforts led to the discovery of promising compound 15, which showed excellent in vitro ASK1 kinase activity and potent inhibitory effects on ASK1 in AP1-HEK293 cells. In a tumor necrosis factor-α (TNF-α)-induced HT-29 intestinal epithelial cell model, compound 15 exhibited a significantly protective effect on cell viability comparable to that of GS-4997; moreover, compound 15 exhibited no obvious cytotoxicity against HT-29 cells at concentrations up to 25 µM. Mechanistic research demonstrated that compound 15 suppresses phosphorylation in the ASK1-p38/JNK signaling pathway in HT-29 cells, and regulates the expression levels of apoptosis-related proteins. Altogether, these results show that compound 15 may serve as a potential candidate compound for the treatment of inflammatory bowel disease (IBD).


Assuntos
Antineoplásicos/farmacologia , Desenho de Fármacos , Indazóis/farmacologia , MAP Quinase Quinase Quinase 5/antagonistas & inibidores , Inibidores de Proteínas Quinases/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/química , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cristalografia por Raios X , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Indazóis/síntese química , Indazóis/química , MAP Quinase Quinase Quinase 5/metabolismo , Simulação de Acoplamento Molecular , Estrutura Molecular , Inibidores de Proteínas Quinases/síntese química , Inibidores de Proteínas Quinases/química , Relação Estrutura-Atividade , Células Tumorais Cultivadas
11.
Biomed Res Int ; 2021: 2471518, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33763469

RESUMO

Objective: Apoptotic events mediated by mitochondrial injury play an important role on the onset of Pemphigus vulgaris (PV). The thioredoxin-2 (Trx2)/apoptosis signal-regulating kinase 1 (ASK1) signaling pathway is considered a key cascade involved on the regulation of mitochondrial injury. Hence, we have investigated the regulatory mechanism of the Trx2/ASK1 signaling in PV-induced mitochondrial injury. Methods: Serum and tissue samples were collected from clinical PV patients to detect the oxidative stress factors, cell apoptosis, and expression of members from Trx2/ASK1 signaling. HaCaT cells were cultured with the serum of PV patients and transfected with Trx2 overexpression or silencing vector. Changes in the levels of reactive oxygen species (ROS), mitochondrial membrane potential (△ψm), and apoptosis were further evaluated. A PV mouse model was established and administered with Trx2-overexpressing plasmid. The effect of ectopic Trx2 expression towards acantholysis in PV mice was observed. Results: A series of cellular and molecular effects, including (i) increased levels of oxidative stress products, (ii) destruction of epithelial cells in the skin tissues, (iii) induction of apoptosis in keratinocytes, (iv) reduction of Trx2 protein levels, and (v) enhanced phosphorylation of ASK1, were detected in PV patients. In vitro experiments confirmed that Trx2 can inhibit ASK1 phosphorylation, alleviate ROS release, decrease △ψm, and lower the apoptotic rate. Injection of Trx2-overexpressing vectors in vivo could also relieve acantholysis and blister formation in PV mice. Conclusion: The Trx2/ASK1 signaling pathway regulates the incidence of PV mediated by mitochondrial injury.


Assuntos
Queratinócitos/metabolismo , MAP Quinase Quinase Quinase 5/metabolismo , Sistema de Sinalização das MAP Quinases , Mitocôndrias/metabolismo , Proteínas Mitocondriais/metabolismo , Estresse Oxidativo , Pênfigo/metabolismo , Tiorredoxinas/metabolismo , Linhagem Celular , Humanos , Queratinócitos/patologia , Potencial da Membrana Mitocondrial , Mitocôndrias/patologia , Pênfigo/patologia , Espécies Reativas de Oxigênio/metabolismo
12.
Biochem Biophys Res Commun ; 548: 104-111, 2021 04 09.
Artigo em Inglês | MEDLINE | ID: mdl-33640602

RESUMO

Alzheimer's disease (AD) is a neurodegenerative disease with a complicated pathogenesis. F-box and WD-40 domain protein 11 (FBXW11), as a component of the SCF (Skp1-Cul1-F-box) E3 ubiquitin ligase complex, regulates multiple different signaling pathways. However, the effects of FBXW11 on AD progression and the underlying mechanisms have not been studied. In this study, we found that FBXW11 expression was markedly increased in microglial cells stimulated by amyloid-ß (Aß). Immunofluorescence staining showed that FBXW11 was co-localized with Iba-1 in microglial cells, suggesting its potential in regulating neuroinflammation. Meanwhile, significantly elevated expression of FBXW11 was detected in hippocampus of AD mouse models. Then, our in vitro studies showed that FBXW11 deletion considerably ameliorated inflammatory response in Aß-incubated microglial cells through suppressing nuclear transcription factor κB (NF-κB) signaling. We further found that FBXW11 physically interacted with apoptosis signal-regulating kinase 1 (ASK1) and promoted its ubiquitination, which led to the aberrant activation of NF-κB and mitogen-activated protein kinase (MAPK) signaling pathways. Importantly, promoting ASK1 significantly abolished the effects of FBXW11 knockdown to repress inflammation and MAPKs/NF-κB activation in Aß-treated microglial cells. Subsequently, our in vivo experiments demonstrated that hippocampus-specific knockout of FBXW11 dramatically alleviated Aß plaque load, neuronal death, and microglial activation in AD mice. Furthermore, hippocampal deficiency of FBXW11 markedly mitigated neuroinflammation in AD mice through restraining ASK1/MAPKs/NF-κB signaling, along with alleviated cognitive deficits. Together, our findings demonstrated that FBXW11 may be a functionally important mediator of ASK1 activation, which could be a novel molecular target for AD treatment.


Assuntos
Peptídeos beta-Amiloides/metabolismo , Deleção de Genes , Inflamação/patologia , MAP Quinase Quinase Quinase 5/metabolismo , Placa Amiloide/patologia , Transdução de Sinais , Proteínas Contendo Repetições de beta-Transducina/deficiência , Doença de Alzheimer/complicações , Doença de Alzheimer/metabolismo , Animais , Linhagem Celular , Transtornos Cognitivos/complicações , Transtornos Cognitivos/metabolismo , Humanos , Sistema de Sinalização das MAP Quinases , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Microglia/metabolismo , NF-kappa B/metabolismo , Placa Amiloide/complicações , Ubiquitinação , Proteínas Contendo Repetições de beta-Transducina/metabolismo
13.
Life Sci ; 272: 119267, 2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-33631173

RESUMO

AIMS: Indoxyl sulfate (IS), a protein-bound uremic toxin, is implicated in endothelial dysfunction, which contributes to adverse cardiovascular events in chronic kidney disease. Apoptosis signal regulating kinase 1 (ASK1) is a reactive oxygen species-driven kinase involved in IS-mediated adverse effects. This study assessed the therapeutic potential of ASK1 inhibition in alleviating endothelial effects induced by IS. MAIN METHODS: IS, in the presence and absence of a selective ASK1 inhibitor (GSK2261818A), was assessed for its effect on vascular reactivity in rat aortic rings, and cultured human aortic endothelial cells where we evaluated phenotypic and mechanistic changes. KEY FINDINGS: IS directly impairs endothelium-dependent vasorelaxation and endothelial cell migration. Mechanistic studies revealed increased production of reactive oxygen species-related markers, reduction of endothelial nitric oxide synthase and increased protein expression of tissue inhibitor of matrix metalloproteinase 1 (TIMP1). IS also increases angiopoietin-2 and tumour necrosis factor α gene expression and promotes transforming growth factor ß receptor abundance. Inhibition of ASK1 ameliorated the increase in oxidative stress markers, promoted autocrine interleukin 8 pro-angiogenic signalling and decreased anti-angiogenic responses at least in part via reducing TIMP1 protein expression. SIGNIFICANCE: ASK1 inhibition attenuated vasorelaxation and endothelial cell migration impaired by IS. Therefore, ASK1 is a viable intracellular target to alleviate uremic toxin-induced impairment in the vasculature.


Assuntos
Endotélio/metabolismo , MAP Quinase Quinase Quinase 5/antagonistas & inibidores , MAP Quinase Quinase Quinase 5/metabolismo , Animais , Apoptose/efeitos dos fármacos , Células Cultivadas , Células Endoteliais/metabolismo , Endotélio/efeitos dos fármacos , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/metabolismo , Indicã/efeitos adversos , Indicã/farmacologia , MAP Quinase Quinase Quinase 5/fisiologia , Masculino , NADPH Oxidases/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Vasodilatação/efeitos dos fármacos
14.
Ecotoxicol Environ Saf ; 213: 112066, 2021 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-33610944

RESUMO

As an emerging pollutant in the aquatic environment, microcystin-LR (MC-LR) can enter the body through multiple pathways, and then induce apoptosis and gonadal damage, affecting reproductive function. Previous studies focused on male reproductive toxicity induced by MC-LR neglecting its effects on females. The apoptotic signal-regulated kinase 1 (ASK1) is an upstream protein of P38/JNK pathway, closely associated with apoptosis and organ damage. However, the role of ASK1 in MC-LR-induced reproductive toxicity is unclear. Therefore, this study investigated the role of ASK1 in mouse ovarian injury and apoptosis induced by MC-LR. After MC-LR exposure, ASK1 expression in mouse ovarian granulosa cells was increased at the protein and mRNA levels, and decreased following pretreatment by antioxidant N-acetylcysteine, suggesting that MC-LR-induced oxidative stress has a regulatory role in ASK1 expression. Inhibition of ASK1 expression with siASK1 and NQDI-1 could effectively alleviate MC-LR-induced mitochondrial membrane potential damage and apoptosis in ovarian granulosa cells, as well as pathological damage, apoptosis and the decreased gonadal index in ovaries of C57BL/6 mice. Moreover, the P38/JNK pathway and downstream apoptosis-related proteins (P-P38, P-JNK, P-P53, Fas) and genes (MKK4, MKK3, Ddit3, Mef2c) were activated in vivo and vitro, but their activation was restrained after ASK1 inhibition. Data presented herein suggest that the ASK1-mediated P38/JNK pathway is involved in ovarian injury and apoptosis induced by MC-LR in mice. It is confirmed that ASK1 has an important role in MC-LR-induced ovarian injury, which provides new insights for preventing MCs-induced reproductive toxicity in females.


Assuntos
Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Toxinas Marinhas/toxicidade , Microcistinas/toxicidade , Animais , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose , Feminino , MAP Quinase Quinase Quinase 5/metabolismo , Sistema de Sinalização das MAP Quinases/fisiologia , Masculino , Potencial da Membrana Mitocondrial , Camundongos , Camundongos Endogâmicos C57BL , Ovário
15.
Medicine (Baltimore) ; 100(3): e23986, 2021 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-33545988

RESUMO

ABSTRACT: Rosacea is a facial chronic inflammatory skin disease with immune and vascular system dysfunction. Paeoniflorin (PF) is a traditional Chinese medicine with anti-inflammatory properties. However, its effects on rosacea remain unknown. Here, we investigated the mechanisms through which PF inhibits the macrophage-related rosacea-like inflammatory response. Immunohistochemical methods were used to detect differences in the inflammatory response and degree of macrophage infiltration in granulomatous rosacea lesions and their peripheral areas. Cell Counting Kit-8 was used to determine the cytotoxicity of PF towards RAW 264.7 cells. Reverse transcription-quantitative polymerase chain reaction and western blotting were used to measure the influence of PF on mRNA and protein expression levels of suppressor of cytokine signaling 3 (SOCS3), apoptosis signal-regulating kinase 1 (ASK1)-p38, Toll-like receptor 2, and cathelicidin antimicrobial peptide ( or LL37) in the lipopolysaccharide (LPS)-induced macrophage-related rosacea-like inflammatory response of RAW 264.7 cells. Inflammatory cell infiltration was more pronounced in granulomatous rosacea lesions than in peripheral areas. LL37 expression increased significantly, and the infiltration of a large number of CD68+ macrophages was observed in the lesions. PF promoted SOCS3 expression in RAW 264.7 cells and inhibited the LPS-induced increase in toll-like receptor 2 and LL37 expression through the ASK1-p38 cascade, thereby alleviating the macrophage-related rosacea-like inflammatory response. These changes could be abrogated by SOCS3 siRNA in vitro.In conclusion, the pathogenesis of rosacea involves abnormal macrophage infiltration within the lesions. PF inhibits the macrophage-related rosacea-like inflammatory response through the SOCS3-ASK1-p38 pathway, demonstrating its potential application as a novel drug for rosacea therapy.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Glucosídeos/farmacologia , MAP Quinase Quinase Quinase 5/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Monoterpenos/farmacologia , Rosácea/tratamento farmacológico , Proteína 3 Supressora da Sinalização de Citocinas/metabolismo , Animais , Técnicas de Cultura de Células , Humanos , Macrófagos/metabolismo , Camundongos , Células RAW 264.7 , Pele/citologia
16.
Cancer Sci ; 112(4): 1633-1643, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33565179

RESUMO

Tumor metastasis is the leading cause of death worldwide and involves an extremely complex process composed of multiple steps. Our previous study demonstrated that apoptosis signal-regulating kinase 1 (ASK1) deficiency in mice attenuates tumor metastasis in an experimental lung metastasis model. However, the steps of tumor metastasis regulated by ASK1 remain unclear. Here, we showed that ASK1 deficiency in mice promotes natural killer (NK) cell-mediated intravascular tumor cell clearance in the initial hours of metastasis. In response to tumor inoculation, ASK1 deficiency upregulated immune response-related genes, including interferon-gamma (IFNγ). We also revealed that NK cells are required for these anti-metastatic phenotypes. ASK1 deficiency augmented cytokine production chemoattractive to NK cells possibly through induction of the ligand for NKG2D, a key activating receptor of NK cells, leading to further recruitment of NK cells into the lung. These results indicate that ASK1 negatively regulates NK cell-dependent anti-tumor immunity and that ASK1-targeted therapy can provide a new tool for cancer immunotherapy to overcome tumor metastasis.


Assuntos
Células Matadoras Naturais/imunologia , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , MAP Quinase Quinase Quinase 5/metabolismo , Metástase Neoplásica/patologia , Animais , Linhagem Celular , Linhagem Celular Tumoral , Feminino , Células HEK293 , Humanos , Imunoterapia/métodos , Interferon gama/metabolismo , Células Matadoras Naturais/metabolismo , Pulmão/imunologia , Pulmão/metabolismo , Pulmão/patologia , Neoplasias Pulmonares/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Metástase Neoplásica/imunologia , Células RAW 264.7
17.
Mol Biochem Parasitol ; 242: 111364, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33639230

RESUMO

Inhibition of an imperative antioxidant enzyme with subsequent death is a victorious and widely accepted strategy to combat various infectious diseases. Among different antioxidant enzymes, thioredoxin reductase (TrxR) is an exclusive one. Studies have revealed that direct inhibition of TrxR by different classes of chemical moieties promptly results in the death of an organism. Especially the structural as well as biochemical modifications of the enzyme upon inhibition project serious threat towards the subject organism. Herein, an attempt was made to inhibit TrxR of filarial species by administering Auranofin, 1 chloro 2,4 dinitrobenzene (CDNB), Curcumin, and a novel carbamo dithioperoxo(thioate) derivative (4a). Our study has revealed that inhibition of TrxR resulted in the induction of the classical CED pathway of apoptosis along with the intrinsic and extrinsic pathways of apoptosis (Caspase mediated) routed through the ASK-1/p38 axis. Druggability analysis of filarial TrxR for the selected compounds was performed in silico through molecular docking studies. Therefore, this study attempts to decipher the mechanism of apoptosis induction following TrxR inhibition. The safety of those four compounds in terms of dose and toxicity was taken under consideration. Thitherto, the mechanism of TrxR mediated initiation of cell death in filarial parasite has remained undercover, and therefore, it is a maiden report on the characterization of apoptosis induction upon TrxR inhibition which will eventually help in generating effective antifilarial drugs in the future.


Assuntos
Anti-Helmínticos/farmacologia , Auranofina/farmacologia , Caspases/genética , Curcumina/farmacologia , Dinitroclorobenzeno/farmacologia , Setaria (Nematoide)/efeitos dos fármacos , Tiorredoxina Dissulfeto Redutase/antagonistas & inibidores , Animais , Anti-Helmínticos/química , Apoptose/efeitos dos fármacos , Apoptose/genética , Auranofina/química , Sítios de Ligação , Caspases/metabolismo , Bovinos , Curcumina/química , Dinitroclorobenzeno/química , Regulação da Expressão Gênica , Proteínas de Helminto/genética , Proteínas de Helminto/metabolismo , MAP Quinase Quinase Quinase 5/genética , MAP Quinase Quinase Quinase 5/metabolismo , Microfilárias/efeitos dos fármacos , Microfilárias/enzimologia , Microfilárias/crescimento & desenvolvimento , Modelos Moleculares , Estresse Oxidativo , Ligação Proteica , Conformação Proteica em alfa-Hélice , Conformação Proteica em Folha beta , Domínios e Motivos de Interação entre Proteínas , Espécies Reativas de Oxigênio/agonistas , Espécies Reativas de Oxigênio/metabolismo , Setaria (Nematoide)/enzimologia , Setaria (Nematoide)/crescimento & desenvolvimento , Transdução de Sinais , Tiorredoxina Dissulfeto Redutase/química , Tiorredoxina Dissulfeto Redutase/genética , Tiorredoxina Dissulfeto Redutase/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
18.
J Cell Biol ; 220(2)2021 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-33399853

RESUMO

Invadosomes support cell invasion by coupling both acto-adhesive and extracellular matrix degradative functions, which are apparently antagonistic. ß1-integrin dynamics regulate this coupling, but the actual sensing mechanism and effectors involved have not yet been elucidated. Using genetic and reverse genetic approaches combined with biochemical and imaging techniques, we now show that the calcium channel TRPV4 colocalizes with ß1-integrins at the invadosome periphery and regulates its activation and the coupling of acto-adhesive and degradative functions. TRPV4-mediated regulation of podosome function depends on its ability to sense reactive oxygen species (ROS) in invadosomes' microenvironment and involves activation of the ROS/calcium-sensitive kinase Ask1 and binding of the motor MYO1C. Furthermore, disease-associated TRPV4 gain-of-function mutations that modulate ECM degradation are also implicated in the ROS response, which provides new perspectives in our understanding of the pathophysiology of TRPV4 channelopathies.


Assuntos
Podossomos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Canais de Cátion TRPV/metabolismo , Actinas/metabolismo , Animais , Cálcio/metabolismo , Adesão Celular , Cisteína/metabolismo , Ácido Ditionitrobenzoico , Matriz Extracelular/metabolismo , Células HEK293 , Humanos , Peróxido de Hidrogênio/metabolismo , Integrina beta1/metabolismo , MAP Quinase Quinase Quinase 5/metabolismo , Camundongos , Modelos Biológicos , Miosina Tipo I/metabolismo , Transporte Proteico , Células RAW 264.7
19.
Clin Sci (Lond) ; 135(1): 161-166, 2021 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-33416082

RESUMO

Ischemia-reperfusion injury (IRI) consequent to major liver surgery is a still unmet clinical problem. The activation of endogenous systems of hepatoprotection can prevent the damaging effects of ischemia-reperfusion (IR) as shown by the phenomenon known as 'ischemic preconditioning'. The identification of endogenous signal mediators of hepatoprotection is of main interest since they could be targeted in future therapeutic interventions. Qiu et al. recently reported in Clin. Sci. (Lond.) (2020) 134(17), 2279-2294, the discovery of a novel protective molecule against hepatic IR damage: dual-specificity phosphatase 12 (DUSP12). IR significantly decreased DUSP12 expression in liver whereas DUSP12 overexpression in hepatocytes protected IRI and DUSP12 deletion in DUSP12 KO mice exacerbated IRI. The protective effects of DUSP12 depended on apoptosis signal-regulating kinase 1 (ASK1) and acted through the inhibition of the ASK1-dependent kinases c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK). These results enlighten DUSP12 as a novel intermediate negative regulator of the pro-inflammatory and pro-apoptotic ASK1/JNK-p38 MAPK pathway activated during hepatic IR and identify DUSP12 as potential therapeutic target for IRI.


Assuntos
Fosfatases de Especificidade Dupla/metabolismo , Fígado/patologia , MAP Quinase Quinase Quinase 5/antagonistas & inibidores , Traumatismo por Reperfusão/prevenção & controle , Transdução de Sinais , Animais , Células Endoteliais/metabolismo , Células Endoteliais/patologia , MAP Quinase Quinase Quinase 5/metabolismo , Camundongos
20.
Int J Mol Med ; 47(2): 732-740, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33416127

RESUMO

Diabetic retinopathy (DR) is the leading cause of blindness among the working­age population in several countries. Despite the available treatments, some patients are diagnosed at the late stages of the disease when treatment is more difficult. Hence, it is crucial that novel targets are identified in order to improve the clinical therapy of DR. In the present study, an animal model of DR and a cell model using primary human retinal microvascular endothelial cells exposed to high glucose were constructed to examine the association between apoptosis signal­regulating kinase 1 (ASK1)/p38 and NLR family pyrin domain containing 3 (NLRP3) in DR. The results revealed that DR induced inflammatory response and microvascular cell proliferation. NLRP3 contributed to DR­mediated inflammatory development and progression, which promoted the expression of inflammatory­related cytokines. In addition, NLRP3 promoted the tube formation of retinal microvascular endothelial cells and angiogenesis. Moreover, further research indicated that the NLRP3­mediated aberrant retinal angiogenesis in DR was regulated by ASK1 and p38. It was thus suggested that ASK1/p38 may be novel target for the treatment of DR.


Assuntos
Retinopatia Diabética/metabolismo , MAP Quinase Quinase Quinase 5/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Neovascularização Retiniana/metabolismo , Transdução de Sinais , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Retinopatia Diabética/genética , Retinopatia Diabética/patologia , Humanos , MAP Quinase Quinase Quinase 5/genética , Masculino , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Neovascularização Retiniana/genética , Neovascularização Retiniana/patologia , Proteínas Quinases p38 Ativadas por Mitógeno/genética
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