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1.
Life Sci ; 241: 117146, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31816325

RESUMO

AIMS: Diabetic retinopathy (DR) is the main cause of blindness in adults and investigating new therapeutic targets for DR is necessary. This study aimed to investigate the effect of high-mobility group box 1 (HMGB1) protein and its mechanism in diabetic retinopathy (DR) were investigated. MAIN METHODS: Human retinal endothelial cells (HREC) were uesd for chip-seq. Sprague Dawley (SD) rats were randomly divided into control group, HMGB1 group, diabetes mellitus (DM) combined with HMGB1 siRNA group, and DM group. Next, eyeballs were removed and retinas were detached for western blot. The DM model of cell was built by increasing the glucose concentration in cell culture medium. The regulation of HMGB1 was achieved by short hairpin (sh)-HMGB1 transfection, then, the transfected cells were harvested for luciferase assay, western blot and qRT-PCR analyses as well as proliferation and apoptosis detection. KEY FINDINGS: Chip-seq and luciferase assay showed the possible transcription factor functions of HMGB1 and IKB-α was one of the HMGB1 binding sites. In vivo and in vitro results indicated high expression of HMGB1 and NF-kB and low expression of IKB-α in DR and the expression of IKB-α and NF-kB was regulated by HMGB1. Moreover, cell assays showed that HMGB1 inhibited cell proliferation and promoted apoptosis. SIGNIFICANCE: The results from the present study showed that HMGB1 may be involved in the pathogenesis of DR as a transcription factor through NF-kB pathway. Therefore, blockade of HMGB1 may be a new method for the treatment of DR.


Assuntos
Diabetes Mellitus Experimental/complicações , Retinopatia Diabética/patologia , Modelos Animais de Doenças , Regulação da Expressão Gênica , Proteína HMGB1/metabolismo , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/metabolismo , Animais , Apoptose , Proliferação de Células , Células Cultivadas , Retinopatia Diabética/etiologia , Retinopatia Diabética/metabolismo , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Proteína HMGB1/genética , Humanos , Masculino , Inibidor de NF-kappaB alfa/genética , NF-kappa B/genética , Ratos , Ratos Sprague-Dawley , Retina/citologia , Retina/metabolismo
2.
Molecules ; 24(18)2019 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-31547327

RESUMO

Previous studies have revealed the anti-inflammatory and neuroprotective properties of Hericium erinaceus extracts, including the fact that the active ingredient erinacine C (EC) can induce the synthesis of nerve growth factor. However, there is limited research on the use and mechanisms of action of EC in treating neuroinflammation. Hence, in this study, the inflammatory responses of human BV2 microglial cells induced by LPS were used to establish a model to assess the anti-neuroinflammatory efficacy of EC and to clarify its possible mechanisms of action. The results showed that EC was able to reduce the levels of nitric oxide (NO), interleukin-6 (IL-6), tumor necrosis factor (TNF)-α, and inducible nitric oxide synthase (iNOS) proteins produced by LPS-induced BV2 cells, in addition to inhibiting the expression of NF-κB and phosphorylation of IκBα (p-IκBα) proteins. Moreover, EC was found to inhibit the Kelch-like ECH-associated protein 1 (Keap1) protein, and to enhance the nuclear transcription factor erythroid 2-related factor (Nrf2) and the expression of the heme oxygenase-1 (HO-1) protein. Taken together, these data suggest that the mechanism of action of EC involves the inhibition of IκB, p-IκBα, and iNOS expressions and the activation of the Nrf2/HO-1 pathway.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Diterpenos/farmacologia , Microglia/efeitos dos fármacos , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Heme Oxigenase-1/metabolismo , Interleucina-6/metabolismo , Lipopolissacarídeos/toxicidade , Proteínas de Membrana/metabolismo , Camundongos , Microglia/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Inibidor de NF-kappaB alfa/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
3.
Biomed Pharmacother ; 117: 109155, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31387178

RESUMO

Stroke is a leading cause of mortality and disability globally. Cerebral ischaemia-reperfusion (I/R) injury is characterized by significant inflammation and extensive cell death. Multiple signaling pathways play essential roles in the process, and identifying the unclear crucial regulators of these pathways may provide promising targets for treatment. CASP8 and FADD-like apoptosis regulator (CFLAR) is expressed in multiple organs to regulate inflammation. Here, we reported that CFLAR expression was markedly reduced in brain samples of mice with middle cerebral artery occlusion (MCAO) stroke. Furthermore, CFLAR knockdown markedly elevated the neurological deficit, brain water content and the infarct volume. In addition, significantly promoted inflammation and endoplasmic reticulum (ER) stress was detected in brain tissues of mice after MCAO, as evidenced by the promoted expression of p-IκBα, p-nuclear factor (NF)-κB (p65), glucose-regulated protein 78 (GRP78), PKR-like ER kinase (PERK), activating transcription factor-6 (ATF-6) and cleaved Caspase-12. Notably, MCAO-induced cerebral I/R injury was markedly alleviated in mice over-expressing CFLAR through suppressing inflammation and ER stress. Furthermore, our in vitro results indicated that oxygen-glucose deprivation (OGD)-induced cell death was evidently ameliorated by CFLAR over-expression. In contrast, the cell death triggered by OGD was accelerated by CFLAR knockdown in vitro through enhancing Caspase-3 cleavage, and this effect was obviously ameliorated by the blockage of ER stress using 4-phenyl butyric acid (4-PBA). Collectively, these results demonstrated that CFLAR could be considered as a novel candidate to develop effective therapeutic treatment against cerebral I/R injury.


Assuntos
Isquemia Encefálica/metabolismo , Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/metabolismo , Estresse do Retículo Endoplasmático/fisiologia , Retículo Endoplasmático/metabolismo , Inflamação/metabolismo , Traumatismo por Reperfusão/metabolismo , Animais , Apoptose/fisiologia , Encéfalo/metabolismo , Linhagem Celular , Células HEK293 , Humanos , Infarto da Artéria Cerebral Média/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais/fisiologia , Acidente Vascular Cerebral/metabolismo
4.
Artif Cells Nanomed Biotechnol ; 47(1): 3239-3245, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31364869

RESUMO

Osteoarthritis (OA) is a major public health concern for which a reliable non-invasive treatment option has yet to be developed. In the present study, we investigated the effects of saxagliptin, a novel dipeptidyl peptidase IV (DPP-4) inhibitor, on several important aspects of the pathophysiology of OA using primary human chondrocytes. The results of real-time PCR and ELISA analyses show that saxagliptin treatment significantly decreased mRNA and protein expression of three key cartilage degrading enzymes: matrix metalloproteinase (MMP)-1, MMP-3, and MMP-13. The results of western blot confirmed that this decrease in MMP-1, -3, and -13 expression prevented degradation of type II collagen. We also found that saxagliptin significantly inhibited expression of a disintegrin and metalloproteinase with thrombospondin motif (ADAMTS)-4 and ADAMTS-5, which was reflected by markedly decreased degradation of aggrecan. Inhibition of DPP-4 by saxagliptin also reduced oxidative stress in human primary chondrocytes as evidenced by decreased production of reactive oxygen species (ROS) and increased glutathione (GSH) levels. Additionally, the results of western blot analysis show that the effects of saxagliptin are mediated through the p38/IκBα/NF-κB pathway, which is considered an important treatment target for OA. These findings suggest a potential role for saxagliptin as a novel treatment against OA.


Assuntos
Adamantano/análogos & derivados , Agrecanas/metabolismo , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Colágeno Tipo II/metabolismo , Dipeptídeos/farmacologia , Osteoartrite/tratamento farmacológico , Proteólise/efeitos dos fármacos , Proteína ADAMTS4/metabolismo , Proteína ADAMTS5/metabolismo , Adamantano/farmacologia , Adamantano/uso terapêutico , Condrócitos/patologia , Dipeptídeos/uso terapêutico , Progressão da Doença , Ativação Enzimática/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Produtos Finais de Glicação Avançada/farmacologia , Humanos , Metaloproteinases da Matriz/metabolismo , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/metabolismo , Osteoartrite/metabolismo , Osteoartrite/patologia , Espécies Reativas de Oxigênio/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
5.
Cell Mol Biol (Noisy-le-grand) ; 65(5): 54-58, 2019 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-31304907

RESUMO

Alzheimer's disease (AD) is a high-incidence neurodegenerative disease with complex and diverse pathogenesis. With aging of the population and continuous improvement of living standards, the incidence of AD is on the increase. Therefore, there is need to develop more effective AD drugs in order to improve the quality of life of the elderly. Sakuranetin (SAK) is a dihydroflavonoid compound extracted from plants. It has many physiological properties. In this study, the effect of SAK on spatial discrimination in a rat model of cognitive dysfunction exposed to D-galactose was investigated with respect to its effect on malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels, and on the expressions of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and nuclear factor-κB inhibitory factor-α (IκBα) in hippocampus of rats. The results obtained suggest that SAK may exert protective effects on brain cells through anti-oxidation mechanism. Moreover, the improvement in learning and memory impairment by SAK may also be related to the inhibition of inflammatory mediators in brain tissue. These findings provide scientific evidence that can be exploited for more effective treatment of Alzheimer's disease.


Assuntos
Doença de Alzheimer/tratamento farmacológico , Flavonoides/farmacologia , Hipocampo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Substâncias Protetoras/farmacologia , Animais , Comportamento Animal/efeitos dos fármacos , Disfunção Cognitiva/induzido quimicamente , Modelos Animais de Doenças , Flavonoides/administração & dosagem , Galactose/farmacologia , Glutationa Peroxidase/metabolismo , Hipocampo/metabolismo , Interleucina-6/metabolismo , Masculino , Malondialdeído/metabolismo , Memória/efeitos dos fármacos , Inibidor de NF-kappaB alfa/metabolismo , Oryza/química , Extratos Vegetais/administração & dosagem , Substâncias Protetoras/administração & dosagem , Ratos , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Navegação Espacial/efeitos dos fármacos , Superóxido Dismutase/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
6.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 35(6): 512-517, 2019 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-31292055

RESUMO

Objective To investigate the effect of knockdown of high mobility group protein B1 (HMGB1) on the proliferation of rat mesangial cells (GMCs) cultured in high glucose (HG) and its mechanism. Methods Rat GMCs was cultured and divided into normal group, high glucose treatment group, negative control small interfering RNA combined with high glucose treatment group (siRNA-NC-HG group) and HMGB1 small interference RNA combined with high glucose treatment group (siRNA-HMGB1-HG group). GMCs in the normal group were cultured in normal DMEM medium. GMCs in the HG treatment group were cultured with HG-DMEM medium. The GMCs in the siRNA-HMGB1-HG group, after transfected with siRNA-HMGB1 sequence for 6 hours, were cultured with high glucose medium for 24 hours. GMCs in the siRNA-NC-HG group, after transfected with siRNA-NC sequence for 6 hours, were cultured in HG medium for 24 hours. HMGB1 mRNA expression levels of GMCs were detected by real-time quantitative PCR. MTT assay was used to detect the proliferation of GMCs. Flow cytometry was performed to assess the apoptosis of GMCs. Western blot analysis was used to detect the protein levels of HMGB1, NF-κBp65 and nuclear factor kappa B inhibitor alpha (IκBα). ELISA was used to detect the levels of interleukin-1ß (IL-1ß), IL-6 and tumor necrosis factor α (TNF-α) in the cell supernatants. Results Compared with the siRNA-NC-HG group or HG treatment group, HMGB1 mRNA level decreased in GMCs in the siRNA-HMGB1-HG group, and after 24-, 48-, 72- and 96-hour treatment, the proliferation activity and apoptosis rate of GMCs decreased. After knock-down of HMGB1 level of GMCs, the level of NF-κBp65 protein decreased, the level of IκBα protein increased, and the levels of IL-1ß, IL-6 and TNF-α in the supernatant decreased. Conclusion Knockdown of HMGB1 inhibits proliferation and promotes apoptosis of GMCs induced by HG, which may be related to the inhibition of NF-κB/IκB-α pathway.


Assuntos
Apoptose , Proliferação de Células , Proteína HMGB1/genética , Células Mesangiais/citologia , Animais , Meios de Cultura , Dissacarídeos , Eletrólitos , Técnicas de Silenciamento de Genes , Glucose , Glutamatos , Glutationa , Histidina , Manitol , Inibidor de NF-kappaB alfa/metabolismo , Ratos , Fator de Transcrição RelA/metabolismo
7.
BMC Cancer ; 19(1): 679, 2019 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-31291942

RESUMO

BACKGROUND: Although the prognosis of chronic myeloid leukemia (CML) has dramatically improved, the pathogenesis of CML remains elusive. Studies have shown that sustained phosphorylation of AKT1 plays a crucial role in the proliferation of CML cells. Evidence indicates that in tongue cancer cells, FAM168A, also known as tongue cancer resistance-associated protein (TCRP1), can directly bind to AKT1 and regulate AKT1/NFκB signaling pathways. This study aimed to investigate the role of FAM168A in regulation of AKT1/NFκB signaling pathway and cell cycle in CML. METHODS: FAM168A interference was performed, and the expression and phosphorylation of FAM168A downstream proteins were measured in K562 CML cell line. The possible roles of FAM168A in the proliferation of CML cells were investigated using in vitro cell culture, in vivo animal models and clinical specimens. RESULTS: We found that the expression of FAM168A significantly increased in the peripheral blood mononuclear cells of CML patients, compared with normal healthy controls. FAM168A interference did not change AKT1 protein expression, but significantly decreased AKT1 phosphorylation, significantly increased IκB-α protein level, and significantly reduced nuclear NFκB protein level. Moreover, there was a significant increase of G2/M phase cells and Cyclin B1 level. Immunoprecipitation results showed that FAM168A interacts with breakpoint cluster region (BCR) -Abelson murine leukemia (ABL1) fusion protein and AKT1, respectively. Animal experiments confirmed that FAM168A interference prolonged the survival and reduced the tumor formation in mice inoculated with K562 cells. The results of clinical specimens showed that FAM168A expression and AKT1 phosphorylation were significantly elevated in CML patients. CONCLUSION: This study demonstrates that FAM168A may act as a linker protein that binds to BCR-ABL1 and AKT1, which further mediates the downstream signaling pathways in CML. Our findings demonstrate that FAM168A may be involved in the regulation of AKT1/NFκB signaling pathway and cell cycle in CML.


Assuntos
Proteínas de Fusão bcr-abl/metabolismo , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Subunidade p50 de NF-kappa B/metabolismo , Proteínas/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Animais , Pontos de Checagem do Ciclo Celular , Proliferação de Células , Criança , Ciclina B1/metabolismo , Modelos Animais de Doenças , Feminino , Humanos , Células K562 , Masculino , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Inibidor de NF-kappaB alfa/metabolismo , Fosforilação , Taxa de Sobrevida , Carga Tumoral
8.
Int Immunopharmacol ; 73: 482-490, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31173970

RESUMO

There is currently no specific drug for the treatment of sepsis and antibiotic administration is considered the best option, despite numerous issues. Therefore, the development of drugs to control the pathogen-induced inflammatory responses associated with sepsis is essential. To address this, our study examined the transcriptomes of lipopolysaccharide (LPS)-induced dendritic cells (DCs), identifying TANK-binding kinase1 (Tbk1) as a key factor involved in the inflammatory response. These data suggested drug repositioning of the Tbk1 inhibitor CYT387, currently used for the treatment of myelofibrosis and some cancers, as a candidate for regulating the LPS-induced inflammatory response. CYT387 also inhibited pro-inflammatory cytokine and surface molecule expression by mature DCs after LPS exposure. These effects correlated with both Akt phosphorylation and IκBα degradation. Finally, CYT387 demonstrated therapeutic effects in LPS-induced endotoxemia and Escherichia coli K1-induced mouse models of sepsis and decreased the expression of pro-inflammatory cytokines. In conclusion, our study suggests that drug repositioning of CYT387 may serve as a potential therapeutic for sepsis.


Assuntos
Benzamidas/uso terapêutico , Endotoxemia/tratamento farmacológico , Infecções por Escherichia coli/tratamento farmacológico , Inibidores de Proteínas Quinases/uso terapêutico , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Pirimidinas/uso terapêutico , Sepse/tratamento farmacológico , Animais , Benzamidas/farmacologia , Citocinas/imunologia , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/metabolismo , Reposicionamento de Medicamentos , Endotoxemia/imunologia , Infecções por Escherichia coli/imunologia , Feminino , Lipopolissacarídeos/farmacologia , Camundongos Endogâmicos C57BL , Inibidor de NF-kappaB alfa/metabolismo , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Pirimidinas/farmacologia , Sepse/imunologia , Transcriptoma/efeitos dos fármacos
9.
Biomed Pharmacother ; 116: 108923, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31154269

RESUMO

AIMS: The purpose of this study was to investigate plumbagin (PL) on liver fibrosis in vitro and in vivo and to explore the underlying mechanisms. METHODS: Carbon tetrachloride (CCl4) was used to establish a rat liver fibrosis model, primary hepatic stellate cells (HSCs) were isolated from the rat liver, and fibrosis-related indicators were detected. RESULTS: The results revealed that PL significantly prevented CCl4-induced liver fibrosis, as evidenced by the attenuation of histopathological changes, the decrease of MDA and the increase of SOD and GSH-P X . In addition, PL downregulated the mRNA levels of NOX4 and procollagen I; the protein expression levels of NOX4 and p-IκB; and the transcriptional activity of NF-κB in liver fibrosis rats. Moreover, PL significantly decreased ROS expression, protein expression of α-SMA and collagen III, and activation of NF-κB and inhibited the nuclear translocation of NF-κB p65 in IL-1ß-stimulated HSCs in vitro. CONCLUSION: The results of our study indicate that PL can mitigate liver fibrosis in vitro and in vivo, which may be related to the ROS-mediated NF-кB signaling pathway.


Assuntos
Cirrose Hepática/tratamento farmacológico , Cirrose Hepática/metabolismo , NF-kappa B/metabolismo , Naftoquinonas/uso terapêutico , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Animais , Antioxidantes/metabolismo , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Colágeno Tipo I/metabolismo , Colágeno Tipo III/metabolismo , Células Estreladas do Fígado/efeitos dos fármacos , Células Estreladas do Fígado/metabolismo , Interleucina-1beta/metabolismo , Cirrose Hepática/patologia , Masculino , NADPH Oxidase 4/metabolismo , Inibidor de NF-kappaB alfa/metabolismo , Naftoquinonas/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Fosforilação/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley , Fator de Transcrição RelA/metabolismo
10.
Artif Cells Nanomed Biotechnol ; 47(1): 2139-2145, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31146598

RESUMO

Osteoarthritis (OA) is a common joint disease for which a safe and reliable treatment has yet to be developed. Here, we demonstrated the potential benefit of treatment with paeonol, a derivative of Paeonia suffruticosa, in the treatment and prevention of OA. Chondrogenic cell line ATDC5 cells were cultured with IL-1ß and the effects of paeonol were assessed through qRT-PCR, western blot analysis, MTT, ELISA, and NF-κB luciferase reporter gene assay. Our findings demonstrate a novel ability of paeonol to inhibit numerous factors of OA, including expressions of IL-6, TNF-α, NOX2, PTGS2, NUCB2/nesfatin-1, ICAM-1, VCAM-1, MMP-3/13, degradation of type II collagen, and NF-κB activation through the rescue of IκBα. Additionally, we assessed the effects of paeonol on cell viability to confirm its safety. These findings implicate a valuable potential role of paeonol in the treatment and prevention of OA.


Assuntos
Acetofenonas/farmacologia , Condrócitos/efeitos dos fármacos , Colágeno Tipo II/metabolismo , Interleucina-1beta/farmacologia , Osteoartrite/metabolismo , Osteoartrite/patologia , Proteólise/efeitos dos fármacos , Acetofenonas/uso terapêutico , Proteínas de Ligação ao Cálcio/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Condrócitos/metabolismo , Condrócitos/patologia , Ciclo-Oxigenase 2/metabolismo , Proteínas de Ligação a DNA/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Humanos , Molécula 1 de Adesão Intercelular/metabolismo , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinase 3 da Matriz/metabolismo , NADPH Oxidase 2/metabolismo , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Osteoartrite/tratamento farmacológico , Fosforilação/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Molécula 1 de Adesão de Célula Vascular/metabolismo
11.
Zygote ; 27(3): 187-189, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31182173

RESUMO

SummaryWe report here the existence of bands of higher molecular weight after western blot analysis in three proteins - Skp1, p27 and IκBα in bovine preimplantation embryos. This finding is specific to preimplantation embryos (from the 2-cell stage to the blastocyst stage) and not differentiated fibroblast cells in which these bands were of expected molecular weight. We suggest that these bands of higher molecular weight represent a complex of proteins that are characteristic of preimplantation embryos.


Assuntos
Blastocisto/metabolismo , Desenvolvimento Embrionário , Proteínas/metabolismo , Animais , Blastocisto/citologia , Western Blotting , Bovinos , Inibidor de Quinase Dependente de Ciclina p27/química , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Feminino , Peso Molecular , Inibidor de NF-kappaB alfa/química , Inibidor de NF-kappaB alfa/metabolismo , Proteínas/química , Proteínas Quinases Associadas a Fase S/química , Proteínas Quinases Associadas a Fase S/metabolismo
12.
Biomed Pharmacother ; 117: 109088, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31202173

RESUMO

The acute kidney injury(AKI) caused by nephrotoxic drugs contributes to inflammation and oxidative injury in podocytes. Wedelolactone (WED), a natural compound, is found with activities as anti-inflammation, anti-oxidative, anti-free radical,and etc. In this present study, MPC-5 cells were exposed to the nephrotoxic drugs doxorubicin (DOX). The results showed that WED significantly increased the SOD activity, CAT and GSH-Px levels, while significantly decreased the MDA content and ROS levels in DOX-induced MPC-5 cells. WED could also significantly decrease the levels of cytokines IL-6, MCP-1, TNF-α, and TGF-ß1. Additionally, the activation and phosphorylation of IκKα, IκBα and NF-κB p65 was inhibited by WED. The co-treatment of PDTC (NF-κB inhibitor) and WED significantly reduced NF-κB p65 phosphorylation. These findings suggested that WED alleviated inflammation and oxidative stress of doxorubicin-induced MPC-5 cells through IκK/IκB/NF-κB signaling pathway.


Assuntos
Cumarínicos/farmacologia , Doxorrubicina/farmacologia , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Estresse Oxidativo/efeitos dos fármacos , Podócitos/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Animais , Linhagem Celular , Proteínas I-kappa B/metabolismo , Inflamação/metabolismo , Camundongos , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/metabolismo , Fosforilação/efeitos dos fármacos , Podócitos/metabolismo , Ratos , Fator de Transcrição RelA/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
13.
Eur J Pharmacol ; 858: 172451, 2019 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-31202806

RESUMO

Attachment of monocytes to endothelial cells is a major event in the pathogenesis of atherosclerosis and cardiovascular disease. As atherosclerosis is considered to be an inflammatory disease, increased expression of proinflammatory cytokines greatly contributes to endothelial dysfunction and atherogenesis. Additionally, attachment of monocytes to endothelial cells triggered by cellular adhesion molecules such as vascular cellular adhesion molecule 1 (VCAM-1) and E-selectin plays a vital role in the development of atherosclerotic plaques. Zinc therapy has been suggested as a potential strategy for countering atherosclerosis. In the present study, for the first time to our knowledge, we investigated the potential role of the GPR39 zinc-sensing receptor in mediating the adhesion of monocytes to endothelial cells, oxidative stress and inflammation in human aortic endothelial cells induced by oxidized low-density lipoprotein (ox-LDL). Our findings show that agonism of GPR39 by the selective agonist TC-G 1008 potently reversed the effects of ox-LDL including increased expression of proinflammatory cytokines and chemokines, markers of oxidative stress, and enhanced expression of cellular adhesion molecules. Importantly, we also show that this protective effect is mediated through the nuclear factor-κB (NF-κB) pathway. Taken together, our findings suggest a potential role of GPR39 as a novel therapeutic target for the treatment and prevention of atherosclerosis induced by ox-LDL.


Assuntos
Adesão Celular/efeitos dos fármacos , Células Endoteliais/citologia , Lipoproteínas LDL/farmacologia , Monócitos/citologia , Monócitos/efeitos dos fármacos , Pirimidinas/farmacologia , Receptores Acoplados a Proteínas-G/agonistas , Sulfonamidas/farmacologia , Linhagem Celular Tumoral , Citocinas/metabolismo , Selectina E/metabolismo , Células Endoteliais/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Monócitos/metabolismo , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Fosforilação/efeitos dos fármacos , Molécula 1 de Adesão de Célula Vascular/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
14.
Food Funct ; 10(7): 4001-4009, 2019 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-31210194

RESUMO

Cinnamaldehyde is a flavor isolated from the bark of Cinnamomum cassia Presl and exerts anti-inflammation effects in various diseases. In our study, we investigated the protective effects and the potential mechanism of cinnamaldehyde on atherosclerosis (AS) by using a high fat diet (HFD)-fed ApoE-/- atherosclerotic mouse model. Here, we found that the serum LDL-C, TG and TC levels were elevated and the HDL-C level was decreased in HFD-fed ApoE-/- mice. Cinnamaldehyde treatment significantly decreased inflammatory cytokine (TNF-α, IL-6, NO and MCP-1) overproduction and the serum lipid level. Meanwhile, cinnamaldehyde increased the HDL-C level and down-regulated the activity of lipid peroxidation product MDA in serum. Moreover, cinnamaldehyde reduced the atherosclerotic plaque area in ApoE-/- mice. Furthermore, cinnamaldehyde reduced matrix metalloproteinase-2 (MMP-2) expression and attenuated the high phosphorylation level of IκBα and p65 NF-κB. Overall, our study indicates that cinnamaldehyde may achieve the anti-atherosclerotic effect via the IκB/NF-κB signaling pathway.


Assuntos
Acroleína/análogos & derivados , Apolipoproteínas E/genética , Aterosclerose/tratamento farmacológico , Dieta Hiperlipídica/efeitos adversos , Proteínas I-kappa B/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais/efeitos dos fármacos , Acroleína/farmacologia , Acroleína/uso terapêutico , Animais , Anti-Inflamatórios/farmacologia , Aorta Torácica/patologia , Aterosclerose/patologia , HDL-Colesterol/sangue , LDL-Colesterol/sangue , Cinnamomum aromaticum/química , Citocinas/metabolismo , Modelos Animais de Doenças , Masculino , Malondialdeído/sangue , Metaloproteinase 2 da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Inibidor de NF-kappaB alfa/metabolismo , Óxido Nítrico/sangue , Fosforilação , Extratos Vegetais/farmacologia , Placa Aterosclerótica/tratamento farmacológico , Fator de Transcrição RelA/metabolismo
15.
Gene ; 706: 124-130, 2019 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-31077735

RESUMO

In this study, we constructed a tumor necrosis factor α (TNF-α)-induced synovial cell inflammatory model using human synoviocytes (HS) cell line to explore the function of miR-98 in rheumatoid arthritis (RA). miR-98 mimics or miR-98 inhibitor were transfected into HS cells to up-regulate or down-regulate the expression of miR-98. The proliferation and apoptosis of HS cells were determined using CCK8 assay and flow cytometry, respectively. TargetScan website was utilized to predict the targets of miR-98. Luciferase assay was carried out to verify that IL-10 is a target of miR-98. Western blot was performed to analyze the expression of IL-10, apoptosis-related and NF-κB signaling pathway-related proteins. Our results demonstrated that the expression of miR-98 was up-regulated in HS cells stimulated by TNF-α. Down-regulation of miR-98 by inhibitor in TNF-α-stimulated HS cells dramatically inhibited cell proliferation and promoted cell apoptosis compared with the miR-98 inhibitor NC group. The protein expression of Bcl-2 was declined while the levels of Bax and Bim were increased by miR-98 inhibitor in TNF-α-stimulated HS cells. IL-10 was predicted and verified as a target of miR-98. qRT-PCR and western blot results revealed that the level of IL-10 was negatively regulated by miR-98. Finally, we identified that down-regulation of miR-98 reduced the expression level of p-p65 and p-IκBα in TNF-α-stimulated HS cells. In summary, our present study demonstrated that down-regulation of miR-98 inhibited the proliferation and promoted the apoptosis of TNF-α-stimulated HS partly by targeting IL-10 and regulating NF-κB signaling pathway, insinuating miR-98 as a candidate biomarker in RA.


Assuntos
Apoptose/genética , MicroRNAs/genética , MicroRNAs/fisiologia , Artrite Reumatoide/genética , Linhagem Celular , Proliferação de Células/fisiologia , Fibroblastos/metabolismo , Humanos , Interleucina-10/genética , Interleucina-10/metabolismo , MicroRNAs/metabolismo , Modelos Biológicos , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais/fisiologia , Membrana Sinovial/metabolismo , Sinoviócitos/metabolismo , Sinoviócitos/fisiologia , Fator de Transcrição RelA/metabolismo , Ativação Transcricional , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/fisiologia , Regulação para Cima
16.
PLoS One ; 14(5): e0217065, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31100090

RESUMO

Because little is known about the role of corticotropin-releasing factor (CRF) agonists in regulating responses in pancreatitis, we evaluated the effects of urocortin 2 (UCN2) and stressin1 in caerulein-induced acute pancreatitis (AP) model in rats. Male rats were pretreated with UCN2 or stressin1 for 30 min followed by induction of AP with supraphysiologic doses of caerulein. Serum amylase and lipase activity, pancreatic tissue necrosis, immune cell infiltrate, nuclear factor (NF)-κB activity, trypsin levels, and intracellular Ca2+ ([Ca2+]i) were ascertained. UCN2, but not stressin1 attenuated the severity of AP in rats. UCN2, but not stressin1, reduced serum amylase and lipase activity, cell necrosis and inflammatory cell infiltration in AP. NF-κB activity in pancreatic nuclear extracts increased in AP and UCN2 treatment reduced caerulein-induced increases in NF-κB activity by 42%. UCN2 treatment prevented caerulein-induced degradation of IκB-α in the cytosolic fraction as well as increased levels of p65 subunit of NF-κB in the cytosolic fraction. Pancreatic UCN2 levels decreased in AP compared with saline. UCN2 evoked [Ca2+]i responses in primary acinar cells and abolished caerulein-evoked [Ca2+]i responses at 0.1nM, and decreased by ~50% at 1.0nM caerulein. UCN2 stimulation resulted in redistribution of a portion of F-actin from the apical to the basolateral pole. UCN2 prevented the massive redistribution of F-actin observed with supraphysiologic doses of caerulein. UCN2, but not stressin1 attenuated severity of an experimental pancreatitis model. The protective effects of UCN2, including anti-inflammatory and anti-necrotic effects involve activation of the CRF2 receptor, [Ca2+]i signaling, and inhibition of NF-κB activity.


Assuntos
Ceruletídeo/toxicidade , Hormônio Liberador da Corticotropina/administração & dosagem , Pancreatite/prevenção & controle , Urocortinas/administração & dosagem , Células Acinares/metabolismo , Células Acinares/patologia , Animais , Hormônio Liberador da Corticotropina/genética , Hormônio Liberador da Corticotropina/metabolismo , Masculino , Inibidor de NF-kappaB alfa/genética , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/genética , NF-kappa B/metabolismo , Pancreatite/induzido quimicamente , Pancreatite/patologia , Ratos , Ratos Sprague-Dawley , Transdução de Sinais , Urocortinas/genética , Urocortinas/metabolismo
17.
Clin Lab ; 65(5)2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-31115225

RESUMO

BACKGROUND: The role of C-X-C motif chemokine receptor 1 (CXCR1), inhibitor of kappa B (IκBα), and hypoxia-inducible factor 1 alpha (HIF-1α) have been reported to promote tumorigenesis and progression in colorectal cancer (CRC). This study is to evaluate the expression of CXCR1, IκBα, and hypoxia-inducible factor 1 alpha (HIF-1α), in combination, in CRC tissues. It also aims to analyze the relationship of these three factors with clinico-pathological characteristics. METHODS: CRC and tumor-adjacent tissues were surgically collected from CRC patients. All patients were diagnosed by pathological examination, and none of the patients had received preoperative chemotherapy or radiotherapy. RNA extraction and cDNA synthesis were performed, and the transcription of CXCR1, IκBα, HIF-1α, and ß-actin was quantified by RT-qPCR. RESULTS: The significant increase of CXCR1, HIF-1α, and decrease of IκBα mRNA expression level were observed in tumor samples of CRC patients (p < 0.05). In addition, CXCR1 expression level was correlated with lymph node metastasis (p = 0.013). Also, results demonstrated a relationship between HIF-1α expression and TNM stage and lymph node metastasis (p = 0.047 and p = 0.005, respectively). CXCR1 and HIF-1α simultaneous expression demonstrated the significant relationship with lymph node metastasis in CRC. CONCLUSIONS: Our findings indicated that CXCR1, HIF-1α, and IκBα can be used as potential prognostic factors indicating tumors in the advanced stage in patients with CRC.


Assuntos
Biomarcadores Tumorais/genética , Regulação Neoplásica da Expressão Gênica , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Inibidor de NF-kappaB alfa/genética , Receptores de Interleucina-8A/genética , Idoso , Biomarcadores Tumorais/metabolismo , Feminino , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Inibidor de NF-kappaB alfa/metabolismo , Prognóstico , RNA Mensageiro/genética , Receptores de Interleucina-8A/metabolismo
18.
Ecotoxicol Environ Saf ; 180: 259-268, 2019 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-31096129

RESUMO

Our study determined the toxic effects of zinc oxide (ZnO) particles with different diameters on dopaminergic (DA) neurons, the role of ubiquitin C-terminal hydrolase L1 (UCH-L1) for ZnO particles-induced neurotoxicity, and corresponding molecular mechanisms. We constructed an in vitro cell injury model for DA neurons to analyze the cytotoxicity of ZnO particles using SH-SY5Y cells. Following cell viability assays and flow cytometry, we found that the cytotoxicity of ZnO particles was affected by particle size, time, and dose of exposure. For example, the toxicity of ZnO particles with 50 nm or 100 nm diameter was stronger than that of ZnO particles with 1000 nm diameter. Furthermore, ZnO particles exposure resulted in a significant decrease in UCH-L1 expression in SH-SY5Y; whereas UCH-L1 overexpression led to a significant increase in cell viability and a sharp decrease in ROS level. Western blotting and adenovirus transfection found that exposure to ZnO particles with different diameters all activate the NF-κB signaling in SH-SY5Y cells; whereas UCH-L1 over-expression resulted in increased levels of IκBα, an endogenous inhibitor of NF-κB signaling pathway. ZnO particles with different diameters all induced cytotoxicity in DA neurons, which may be related to the free Zn2+ in the suspension. Regarding the neurotoxic effect of ZnO particles, UCH-L1 protects against and/or alleviates neuronal damage, possibly by deubiquitination of the endogenous inhibitor, IκBα, which leads to activation of NF-κB signaling. Therefore, one possible mechanism for ZnO particle-induced neurotoxicity may be mediated via the down-regulation of UCH-L1 expression in DA cells.


Assuntos
Neurônios Dopaminérgicos/efeitos dos fármacos , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/metabolismo , Ubiquitina Tiolesterase/metabolismo , Óxido de Zinco/toxicidade , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Neurônios Dopaminérgicos/metabolismo , Neurônios Dopaminérgicos/patologia , Regulação para Baixo , Humanos , Inibidor de NF-kappaB alfa/genética , Tamanho da Partícula , Transdução de Sinais/efeitos dos fármacos , Propriedades de Superfície , Óxido de Zinco/química
19.
Artif Cells Nanomed Biotechnol ; 47(1): 1628-1634, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31010320

RESUMO

Chronic inflammation in fibroblast-like synoviocytes (FLSs) induced by pro-inflammatory cytokines such as TNF-α plays a key role in the pathogenesis of rheumatoid arthritis (RA). The neurokinin-1 receptor (NK-1R) is one of the G protein-coupled receptors (GPCRs) mediating the intracellular signalling of substance P (SP). However, the possible implications of NK-1R in rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs) and the pathogenesis of RA have not yet been reported. In the current study, we report that NK-1R is expressed in FLSs. Importantly, NK-1R expression was found to be significantly increased in RA-FLSs compared to normal FLSs. Interestingly, we found that treatment with tumour necrosis factor (TNF)-α increased the expression of NK-1R at both the gene and protein levels. Treatment with the NK-1R antagonist aprepitant reduced TNF-α-induced expression of NADPH oxidase 4 (NOX-4) and generation of reactive oxygen species (ROS) in FLSs. Our results also display that blockage of NF-1R using aprepitant inhibited TNF-α-induced expression and secretion of proinflammatory cytokines, including interleukin-1ß (IL-1ß), IL-6, and IL-8. Consistently, aprepitant prevented TNF-α-induced expression of matrix metalloproteinases (MMPs), including matrix metalloproteinase-3 (MMP-3) and matrix metalloproteinase-13 (MMP-13). Mechanistically, our data demonstrate that treatment with aprepitant inhibited TNF-α-induced phosphorylation and degradation of inhibitor of NF-κB (IκBα). Notably, aprepitant attenuated TNF-α-induced nuclear translocation of nuclear factor κB (NF-κB) p65 and reduced luciferase activity of NF-κB in FLSs. The findings implicated a novel function of NK-1R in RA-FLSs. Blockage of NK-1R using its specific antagonist aprepitant might provide a new therapeutic strategy for RA.


Assuntos
Aprepitanto/farmacologia , Artrite Reumatoide/metabolismo , Artrite Reumatoide/patologia , Fibroblastos/patologia , Antagonistas do Receptor de Neuroquinina-1/farmacologia , Receptores da Neurocinina-1/metabolismo , Sinoviócitos/efeitos dos fármacos , Aprepitanto/uso terapêutico , Artrite Reumatoide/tratamento farmacológico , Artrite Reumatoide/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Inflamação/tratamento farmacológico , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinase 3 da Matriz/metabolismo , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/metabolismo , Antagonistas do Receptor de Neuroquinina-1/uso terapêutico , Estresse Oxidativo/efeitos dos fármacos , Receptores da Neurocinina-1/genética , Transdução de Sinais/efeitos dos fármacos , Sinoviócitos/metabolismo , Sinoviócitos/patologia , Fator de Necrose Tumoral alfa/metabolismo
20.
Biomed Pharmacother ; 115: 108868, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30999127

RESUMO

The characteristic feature of obesity and insulin resistance is chronic low-grade inflammation. Nod-Like Receptor Pyrin 3 (NLRP3) inflammasome plays a central role in obesity-induced insulin resistance. However, how does Carbenoxolone (CBX) play its role in ameliorating insulin resistance in peripheral tissues of obese mice induced by high-fat diet (HFD) remains unknown. In our study, we explored the molecular mechanism of CBX in improving insulin resistance in liver and skeletal muscle in mice induced by the HFD. Our results revealed that in the CBX group, a significant decrease in fasting blood glucose, insulin and HOMA-IR score were observed. CBX could attenuate intracellular lipid accumulation and inflammation aggravation in liver and skeletal muscle. Besides, treatment with CBX could significantly reduce expressions of p-IκB-α, p-NF-κB, p-IRS-1, NLRP3 and inflammatory factors, increase expressions of p-PI3K and p-AKT. Therefore, CBX could dramatically improve insulin resistance in liver and skeletal muscle in mice induced by the high-fat diet. In conclusions, we demonstrate that CBX has a significant protective effect on diet-induced obesity in mice. The potential mechanisms include inhibiting IκB-α/NF-κB pathway, restricting the production of NLRP3 inflammasome and other inflammatory factors, reducing the expression of p-IRS-1, increasing the expressions of p-PI3K and p-AKT, thus ameliorating insulin resistance in liver and skeletal muscle of high-fat diet mice. Therefore CBX is an active agent against diet-induced obesity and is given the opportunity for the treatment of obesity related diseases.


Assuntos
Carbenoxolona/farmacologia , Dieta Hiperlipídica/efeitos adversos , Resistência à Insulina , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/metabolismo , Obesidade/induzido quimicamente , Animais , Regulação da Expressão Gênica/efeitos dos fármacos , Inflamação/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Músculo Esquelético , Inibidor de NF-kappaB alfa/genética , NF-kappa B/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Obesidade/complicações , Distribuição Aleatória
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