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1.
J Agric Food Chem ; 67(39): 10871-10879, 2019 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-31517482

RESUMO

This study evaluated the effect of triterpenoids from edible mushroom Poria cocos on intestinal epithelium integrity and revealed the transcriptional regulatory pathways that underpin restorative mechanisms in the gut. Based on computational docking studies, transcriptional activation experiments and glucocorticoid receptor (GR) protein immunofluorescence localization assays in cultured cells, 16α-hydroxytrametenolic acid (HTA) was discovered as a novel GR agonist in this study. HTA ameliorates TNF-α-induced Caco-2 monolayer intestinal epithelial barrier damage and suppressed activation of phosphatidylinositol 3-kinase (PI3K) and protein kinase B (Akt), which attenuated downstream IκB and nuclear factor kappa-B (NF-κB) phosphorylation through GR activation. Moreover, HTA prevented NF-κB translocation into the nucleus and binding to its cis-element and suppressed lipopolysaccharide-induced downstream NO production and pro-inflammatory cytokines at both protein and mRNA expression levels. In conclusion, HTA from P. cocos improves intestinal barrier function through a GR-mediated PI3K/Akt/NF-κB signaling pathway and may be potentially exploited as a supportive dietary therapeutic strategy for restoring gut health.


Assuntos
Mucosa Intestinal/efeitos dos fármacos , NF-kappa B/metabolismo , Fosfatidilinositol 3-Quinase/metabolismo , Extratos Vegetais/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptores de Glucocorticoides/metabolismo , Triterpenos/farmacologia , Wolfiporia/química , Células CACO-2 , Humanos , Proteínas I-kappa B/genética , Proteínas I-kappa B/metabolismo , Mucosa Intestinal/metabolismo , Simulação de Acoplamento Molecular , NF-kappa B/genética , Fosfatidilinositol 3-Quinase/genética , Fosforilação , Extratos Vegetais/química , Proteínas Proto-Oncogênicas c-akt/genética , Receptores de Glucocorticoides/genética , Transdução de Sinais/efeitos dos fármacos , Triterpenos/química , Verduras/química
2.
Mol Med Rep ; 19(3): 2057-2064, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30664158

RESUMO

The present study aimed to investigate the anti­arthritic effects of curculigoside isolated from the rhizome of Curculigo orchioides Gaertn in vivo and in vitro, as well as to determine the potential underlying mechanisms. A rat model of arthritis was induced with type II collagen. Arthritic rats were treated with curculigoside (50 mg/kg) and blood samples were collected to determine serum levels of tumor necrosis factor (TNF)­α, interleukin (IL)­1ß, IL­6, IL­10, IL­12 and IL­17A. Furthermore, indices of the thymus and spleen were determined. The anti­proliferative effects of curculigoside were detected with Cell Counting kit­8 assays in rheumatoid arthritis­derived fibroblast­like synoviocyte MH7A cells. In addition, expression levels of Janus kinase (JAK)1, JAK3, signal transducer and activator of transcription (STAT)3, nuclear factor (NF)­κB p65 and its inhibitor (IκB) were determined by western blotting. The results revealed that curculigoside inhibited paw swelling and arthritis scores in type II collagen­induced arthritic (CIA) rats. Additionally, curculigoside decreased serum levels of TNF­α, IL­1ß, IL­6, IL­10, IL­12 and IL­17A in CIA rats. Curculigoside also significantly inhibited MH7A cell proliferation in a time and concentration­dependent manner. Furthermore, treatment downregulated the expression of JAK1, JAK3 and STAT3, and upregulated cytosolic nuclear factor (NF)­κB p65 and IκB. In conclusion, the results of the present study indicated that curculigoside exhibited significant anti­arthritic effects in vivo and in vitro, and the molecular mechanism may be associated with the JAK/STAT/NF­κB signaling pathway.


Assuntos
Artrite Reumatoide/tratamento farmacológico , Benzoatos/administração & dosagem , Glucosídeos/administração & dosagem , Janus Quinase 1/genética , Janus Quinase 3/genética , Fator de Transcrição STAT3/genética , Animais , Artrite Reumatoide/induzido quimicamente , Artrite Reumatoide/genética , Artrite Reumatoide/patologia , Proliferação de Células/efeitos dos fármacos , Colágeno Tipo II/toxicidade , Modelos Animais de Doenças , Expressão Gênica/efeitos dos fármacos , Humanos , Proteínas I-kappa B/genética , Interleucinas/genética , Ratos , Transdução de Sinais , Sinoviócitos/efeitos dos fármacos , Fator de Transcrição RelA/genética , Fator de Necrose Tumoral alfa/genética
3.
Am J Ophthalmol ; 200: 150-160, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30653959

RESUMO

PURPOSE: To investigate the expression pattern of nucleotide-binding oligomerization domain (Nod)-like receptors that detects "danger" intracellular signaling and its correlation with clinical dry eye (DE) markers. DESIGN: Cross-sectional study. METHODS: A total of 50 participants with 50 eyes were included: 23 eyes with Sjögren syndrome (SS)-DE, 14 eyes with non-SS-DE, and 13 healthy controls with non-DE. Ocular Surface Disease Index (OSDI) was self-answered and clinical tests including the tear film breakup time (TBUT), Schirmer test, and corneal fluorescein staining (CFS) were performed. Specimens for expression pattern analysis were obtained by conjunctival impression cytology and biopsy. Nod-1, inhibitor kappa B kinase-alpha (IκKα), and nuclear factor kappa B (NF-κB) expression was determined by reverse transcription quantitative real-time polymerase chain reaction and Western blot. Correlations between Nod-1 and ocular surface parameters were determined. RESULTS: Patients with SS-DE had significantly higher OSDI and CFS scores and lower TBUT and Schirmer test scores than those with non-SS-DE patients (all P < .05). Compared with the control group, both the SS-DE and non-SS-DE groups showed significant upregulation in mRNA expression levels of Nod-1 (relative 3.48-fold and 1.72-fold upregulation, respectively, P < .01), IκKα (relative 1.83-fold and 1.24-fold upregulation, respectively, P < .01), and NF-κB (relative 1.84-fold and 1.32-fold upregulation, respectively, P < .01). Western blot analysis showed that Nod-1 protein expression increased in both the SS-DE and non-SS-DE groups (relative 2.71-fold and 1.64-fold upregulation, respectively, P < .05) compared with that in the control group. Similar findings were observed for IκKα and NF-κB. In DE participants, the expression of Nod-1 significantly correlated with the OSDI (R2 = 0.61, r = 0.78, P < .01), Schirmer test score (R2 = 0.44, r = -0.66, P < .01), and CFS (R2 = 0.46, r = 0.68, P < .01) but did not significantly correlate with TBUT (R2 < 0.01, r = 0.08, P = .66). CONCLUSIONS: Nod-1 expression was increased in the conjunctiva of DE, especially SS-DE, and was associated with disease severity. Expression of Nod-like receptors might play an important role in initiating the inflammatory response in DE.


Assuntos
Túnica Conjuntiva/metabolismo , Regulação da Expressão Gênica/fisiologia , Proteína Adaptadora de Sinalização NOD1/genética , Síndrome de Sjogren/genética , Idoso , Western Blotting , Estudos Transversais , Feminino , Marcadores Genéticos/genética , Humanos , Proteínas I-kappa B/genética , Peptídeos e Proteínas de Sinalização Intracelular/genética , Masculino , Pessoa de Meia-Idade , NF-kappa B/genética , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Síndrome de Sjogren/diagnóstico
4.
Nat Commun ; 9(1): 4001, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30275490

RESUMO

Diffuse large B-cell lymphoma (DLBCL) is an aggressive cancer originating from mature B-cells. Prognosis is strongly associated with molecular subgroup, although the driver mutations that distinguish the two main subgroups remain poorly defined. Through an integrative analysis of whole genomes, exomes, and transcriptomes, we have uncovered genes and non-coding loci that are commonly mutated in DLBCL. Our analysis has identified novel cis-regulatory sites, and implicates recurrent mutations in the 3' UTR of NFKBIZ as a novel mechanism of oncogene deregulation and NF-κB pathway activation in the activated B-cell (ABC) subgroup. Small amplifications associated with over-expression of FCGR2B (the Fcγ receptor protein IIB), primarily in the germinal centre B-cell (GCB) subgroup, correlate with poor patient outcomes suggestive of a novel oncogene. These results expand the list of subgroup driver mutations that may facilitate implementation of improved diagnostic assays and could offer new avenues for the development of targeted therapeutics.


Assuntos
Regulação Neoplásica da Expressão Gênica , Genes Reguladores/genética , Variação Genética , Genoma Humano/genética , Linfoma Difuso de Grandes Células B/genética , Regiões 3' não Traduzidas/genética , Linfócitos B/metabolismo , Linfócitos B/patologia , Linhagem Celular Tumoral , Exoma/genética , Estudo de Associação Genômica Ampla , Centro Germinativo/metabolismo , Centro Germinativo/patologia , Humanos , Proteínas I-kappa B/genética , Linfoma Difuso de Grandes Células B/metabolismo , Linfoma Difuso de Grandes Células B/patologia , Mutação , Proteínas Nucleares/genética , Receptores de IgG/genética , Análise de Sequência de DNA , Transcriptoma
5.
J Immunol ; 201(7): 1907-1917, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30127089

RESUMO

In both NOD mice and humans, the development of type 1 diabetes (T1D) is dependent in part on autoreactive CD8+ T cells recognizing pancreatic ß cell peptides presented by often quite common MHC class I variants. Studies in NOD mice previously revealed that the common H2-Kd and/or H2-Db class I molecules expressed by this strain aberrantly lose the ability to mediate the thymic deletion of pathogenic CD8+ T cell responses through interactions with T1D susceptibility genes outside the MHC. A gene(s) mapping to proximal chromosome 7 was previously shown to be an important contributor to the failure of the common class I molecules expressed by NOD mice to mediate the normal thymic negative selection of diabetogenic CD8+ T cells. Using an inducible model of thymic negative selection and mRNA transcript analyses, we initially identified an elevated Nfkbid expression variant as a likely NOD-proximal chromosome 7 region gene contributing to impaired thymic deletion of diabetogenic CD8+ T cells. CRISPR/Cas9-mediated genetic attenuation of Nfkbid expression in NOD mice resulted in improved negative selection of autoreactive diabetogenic AI4 and NY8.3 CD8+ T cells. These results indicated that allelic variants of Nfkbid contribute to the efficiency of intrathymic deletion of diabetogenic CD8+ T cells. However, although enhancing thymic deletion of pathogenic CD8+ T cells, ablating Nfkbid expression surprisingly accelerated T1D onset that was associated with numeric decreases in both regulatory T and B lymphocytes in NOD mice.


Assuntos
Linfócitos T CD8-Positivos/imunologia , Cromossomos Humanos Par 7/genética , Diabetes Mellitus Tipo 1/imunologia , Proteínas I-kappa B/genética , Timo/imunologia , Alelos , Animais , Autoantígenos/imunologia , Diferenciação Celular , Células Cultivadas , Deleção Clonal , Modelos Animais de Doenças , Suscetibilidade a Doenças , Humanos , Proteínas I-kappa B/metabolismo , Camundongos , Camundongos Endogâmicos NOD , Polimorfismo Genético
6.
Int J Mol Sci ; 19(6)2018 06 13.
Artigo em Inglês | MEDLINE | ID: mdl-29899297

RESUMO

Primary cutaneous diffuse large B-cell lymphoma, leg type (PCDLBCL-LT) is one of the well-recognized extranodal lymphomas commonly addicted to the B-cell receptor-MYD88 superpathway. We aimed to describe the genomic changes in a patient who progressed through treatment with ibrutinib, a Bruton's tyrosine kinase (BTK) inhibitor. An 80-year-old woman presented with multiply relapsed PCDLBCL-LT after multiple lines of chemoimmunotherapy and radiotherapy. Pre-treatment testing of the localized cutaneous tumor lesion on a lymphoid amplicon panel demonstrated an MYD88 p.L265P mutation. Ibrutinib therapy was subsequently commenced, resulting in complete resolution of the skin disease. Despite an ongoing skin response, the patient developed progressive nodal disease at two months. Genomic analysis of the cutaneous tumor sample at baseline was compared to that of the inguinal lymph node upon progression, and revealed the acquisition of multiple genomic changes. These included several aberrations expected to bypass BTK inhibition, including two CARD11-activating mutations, and a deleterious mutation in the nuclear factor kappa B (NF-κB) negative regulator, NFKBIE. In addition, an IgH-IRF8 translocation was detected (which brings the IRF8 transcription factor under control of the immunoglobulin heavy chain locus), representing a third plausible mechanism contributing to ibrutinib resistance. Several copy-number changes occurred in both samples, including an amplification of 18q, which encodes the anti-apoptotic protein BCL2. We describe the first case of novel genomic changes of PCDLBCL-LT that occurred while on ibrutinib, providing important mechanistic insights into both pathogenesis and drug resistance.


Assuntos
Antineoplásicos/uso terapêutico , Linfoma Difuso de Grandes Células B/genética , Pirazóis/uso terapêutico , Pirimidinas/uso terapêutico , Neoplasias Cutâneas/genética , Idoso de 80 Anos ou mais , Antineoplásicos/administração & dosagem , Proteínas Adaptadoras de Sinalização CARD/genética , Progressão da Doença , Feminino , Instabilidade Genômica , Guanilato Ciclase/genética , Humanos , Proteínas I-kappa B/genética , Fatores Reguladores de Interferon/genética , Metástase Linfática , Linfoma Difuso de Grandes Células B/tratamento farmacológico , Linfoma Difuso de Grandes Células B/patologia , Mutação , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , Pirazóis/administração & dosagem , Pirimidinas/administração & dosagem , Neoplasias Cutâneas/tratamento farmacológico , Neoplasias Cutâneas/patologia
7.
Exp Dermatol ; 27(9): 1048-1052, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29938836

RESUMO

Antagonists of IL-17A and its receptor have proven to be highly effective in the treatment of psoriasis. However, many of the underlying molecular mechanisms involved in the pathogenesis of psoriasis are still to be determined. IκBζ (encoded by the NFKBIZ gene) plays a key role in the development of psoriasis by mediating IL-17A- and IL-17F-driven effects. Both IL-17A and IL-17F expression are increased in lesional psoriatic skin. IL-17A/A and IL-17F/F homodimers as well as the IL-17A/F heterodimer signal through the same receptors. The aim of this study was to characterize the role of the IL-17A/F heterodimer in the regulation of NFKBIZ expression and in the regulation of selected psoriasis-associated genes. We demonstrated that IL-17A/F stimulation of human keratinocytes significantly induced NFKBIZ expression. Moreover, silencing IκBζ by siRNA revealed that IκBζ is a key regulator of IL-17A/F-inducible psoriasis-associated genes, including CCL20, DEFB4, IL-8, CHI3L1 and S100A7. In addition, IL-17A/F-induced NFKBIZ expression was mediated by a mechanism involving the p38 MAPK and NF-κB signalling pathways. In conclusion, we present IκBζ as a novel key regulator of IL-17A/F-driven effects in psoriasis. Thus, antagonists to IL-17A/F or IκBζ may present a targeted approach for treating psoriasis.


Assuntos
Expressão Gênica/efeitos dos fármacos , Proteínas I-kappa B/genética , Interleucina-17/farmacologia , Proteínas Nucleares/genética , Psoríase/genética , Células Cultivadas , Quimiocina CCL20/genética , Proteína 1 Semelhante à Quitinase-3/genética , Inativação Gênica , Humanos , Interleucina-8/genética , Queratinócitos , Sistema de Sinalização das MAP Quinases , NF-kappa B/metabolismo , Proteína A7 Ligante de Cálcio S100/genética , Fator de Necrose Tumoral alfa/farmacologia , beta-Defensinas/genética
9.
Biochem Biophys Res Commun ; 500(4): 944-951, 2018 06 12.
Artigo em Inglês | MEDLINE | ID: mdl-29705698

RESUMO

The pro-inflammatory vasoconstrictor Angiotensin II can cause endothelial dysfunction and is considered to be one of the mediators of atherosclerosis. Our former results demonstrated that polysaccharides derived from the red alga Porphyridium sp. attenuate inflammatory processes by interfering with tumor necrosis factor-alpha-induced inflammation, in human coronary artery endothelial cells. However, the anti-inflammatory effect of these polysaccharides on inflammation processes occurring under Angiotensin II stimulation is yet unknown. Herein, we studied the polysaccharide's anti-inflammatory effect by quantification of inflammatory markers in Angiotensin II- stimulated Human Coronary Artery Endothelial Cells following pre-treatment with polysaccharides. Inflammatory atherosclerotic pathways up-regulated by Angiotensin II, including adhesion molecule expression and nuclear factor kappa-light-chain-enhancer of activated B cells translocation, were significantly attenuated or diminished in cells pre-treated with the polysaccharides. In addition, the polysaccharides increased the antioxidant response elements activity through the nuclear factor-E2-related factor 2- antioxidant protection system. These polysaccharide's promising abilities may be considered as a basis for future use as a therapeutic agent aimed at improving vascular health by attenuation of the inflammatory atherosclerotic process.


Assuntos
Angiotensina II/farmacologia , Anti-Inflamatórios/farmacologia , Células Endoteliais/efeitos dos fármacos , NF-kappa B/genética , Polissacarídeos/farmacologia , Rodófitas/química , Anti-Inflamatórios/isolamento & purificação , Elementos de Resposta Antioxidante/efeitos dos fármacos , Linhagem Celular , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Regulação da Expressão Gênica , Genes Reporter , Humanos , Proteínas I-kappa B/genética , Proteínas I-kappa B/metabolismo , Inflamação , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/metabolismo , Proteína 1 Associada a ECH Semelhante a Kelch/genética , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Luciferases/genética , Luciferases/metabolismo , Modelos Biológicos , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Polissacarídeos/isolamento & purificação , Transdução de Sinais , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Molécula 1 de Adesão de Célula Vascular/genética , Molécula 1 de Adesão de Célula Vascular/metabolismo
10.
Hum Immunol ; 79(6): 494-498, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29601852

RESUMO

The NF-kappaB pathway might play a role in the pathogenesis of renal disease and type 2 diabetes (T2DM). Our aim was to determine whether common polymorphisms in NF-kappaB genes were associated with impaired renal function and T2DM in a cohort of healthy elderly individuals. We studied 487 individuals, all Caucasian and aged 65-85 years. A total of 104 (21%) had impaired renal function (estimated glomerular filtration rate, eGFR < 60) and 146 (30%) were classified as diabetics. The genotypes of 4 common variants were determined through PCR-RFLP or fluorescent capillary electrophoresis. The NFKB1 variants were significantly associated with T2DM: rs7667496 p = 0.01, OR = 1.68; and rs28362491 p = 0.02, OR = 1.67. They remained significantly associated in a multiple logistic regression with age, gender, hypertension, body mass index, and cholesterol. There was a trend toward the association of these variants with eGFR < 60. The two NFKB1 variants were in linkage disequilibrium (D' = -0.86), and homozygous for the two non-risk alleles (rs7667496 CC + rs28362491 II), were significantly more common in the non-diabetics (p = 0.02). In our cohort the NFKB1 variation was an independent risk factor for developing T2DM. Additional studies to confirm this association are of special interest, as well as studies to give a functional explanation to the genetic association.


Assuntos
Diabetes Mellitus Tipo 2/genética , Genótipo , Proteínas I-kappa B/genética , Rim/metabolismo , Inibidor de NF-kappaB alfa/genética , Subunidade p50 de NF-kappa B/genética , Proteínas Nucleares/genética , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Estudos de Coortes , Grupo com Ancestrais do Continente Europeu , Feminino , Estudos de Associação Genética , Predisposição Genética para Doença , Humanos , Rim/patologia , Masculino , Polimorfismo de Nucleotídeo Único , Fatores Sexuais
11.
Food Funct ; 9(3): 1713-1725, 2018 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-29485160

RESUMO

We conducted the present study to determine the gastric injury preventive effects of polyphenols in Kuding tea (KTPs) in Kunming (KM) mice through the inhibition of gastric-acid secretion and the protection of the gastric mucosa. Mice treated with a high concentration of Kuding tea polyphenols (HKTP) had lower serum levels of interleukin-6 (IL-6), IL-12, tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), motilin (MOT), substance P (SP), and endothelin-1 (ET-1), and higher serum levels of somatostatin (SS) and vasoactive intestinal peptide (VIP) than did the mice in the control group. Serum and gastric tissue levels of nitrous oxide (NO), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and GSH were higher in the HKTP-treated mice than in the control mice, but malondialdehyde (MDA) levels were lower in the HKTP-treated mice than in the control mice. The expression of occludin, epidermal growth factor (EGF), EGF receptor (EGFR), vascular endothelial growth factor (VEGF), nuclear factor of κ-light polypeptide gene enhancer in B-cells inhibitor-α (IκBα), Cu/Zn-SOD (cuprozinc-superoxide dismutase), Mn-SOD (manganese-superoxide dismutase), GSH-Px (glutathione peroxidase), neuronal nitric oxide synthase (nNOS), endothelial NOS (eNOS), messenger RNA (mRNA) and protein in gastric tissue was stronger in the HKTP-treated mice than in the control mice, while the expression of p38 mitogen-activated protein kinase (P38MAPK, or p38), nuclear factor κ-light-chain-enhancer of activated B cells (NF-κB), inducible NOS (iNOS), and cyclooxygenase-2 (COX-2) was weaker in the HKTP group than in the control group. And HKTP also could reduce the TNF-α, IL-1ß (interleukin-1 beta), and IL-6 mRNA expression in gastric injury mice. A high performance liquid chromatography (HPLC) assay showed that Kuding tea polyphenols (KTPs) contained chlorogenic acid, cryptochlorogenic acid, and isochlorogenic acids A, B, and C. These constituents contributed to the preventive effects of KTPs on gastric injury. According to these results, KTPs are a kind of active component that have a strong preventive effect on gastric injury.


Assuntos
Camellia sinensis/química , Extratos Vegetais/administração & dosagem , Polifenóis/administração & dosagem , Gastropatias/tratamento farmacológico , Estômago/lesões , Animais , Etanol/efeitos adversos , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/metabolismo , Glutationa Peroxidase/genética , Glutationa Peroxidase/metabolismo , Humanos , Ácido Clorídrico/efeitos adversos , Proteínas I-kappa B/genética , Proteínas I-kappa B/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Masculino , Malondialdeído/metabolismo , Camundongos , Estômago/efeitos dos fármacos , Gastropatias/induzido quimicamente , Gastropatias/genética , Gastropatias/metabolismo , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Chá/química , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo
12.
Nucleic Acids Res ; 46(8): 4256-4270, 2018 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-29471506

RESUMO

The expression of proteins during inflammatory and immune reactions is coordinated by post-transcriptional mechanisms. A particularly strong suppression of protein expression is exerted by a conserved translational silencing element (TSE) identified in the 3' UTR of NFKBIZ mRNA, which is among the targets of the RNA-binding proteins Roquin-1/2 and MCPIP1/Regnase-1. We present evidence that in the context of the TSE MCPIP1, so far known for its endonuclease activity toward mRNAs specified by distinct stem-loop (SL) structures, also suppresses translation. Overexpression of MCPIP1 silenced translation in a TSE-dependent manner and reduced ribosome occupancy of the mRNA. Correspondingly, MCPIP1 depletion alleviated silencing and increased polysomal association of the mRNA. Translationally silenced NFKBIZ or reporter mRNAs were mostly capped, polyadenylated and ribosome associated. Furthermore, MCPIP1 silenced also cap-independent, CrPV-IRES-dependent translation. This suggests that MCPIP1 suppresses a post-initiation step. The TSE is predicted to form five SL structures. SL4 and 5 resemble target structures reported for MCPIP1 and together were sufficient for MCPIP1 binding and mRNA destabilization. Translational silencing, however, required SL1-3 in addition. Thus the NFKBIZ TSE functions as an RNA element in which sequences adjacent to the site of interaction with MCPIP1 and dispensable for accelerated mRNA degradation extend the functional repertoire of MCPIP1 to translational silencing.


Assuntos
Inativação Gênica , Proteínas I-kappa B/genética , Proteínas Nucleares/genética , Biossíntese de Proteínas , Sequências Reguladoras de Ácido Ribonucleico , Ribonucleases/metabolismo , Fatores de Transcrição/metabolismo , Sítios de Ligação , Células HeLa , Humanos , Elongação Traducional da Cadeia Peptídica , Domínios Proteicos , RNA Mensageiro/metabolismo , Receptor EphB3 , Ribonucleases/química , Ribossomos/metabolismo , Fatores de Transcrição/química
13.
Mol Med Rep ; 17(3): 4376-4382, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29344652

RESUMO

Chrysophanol is an anthraquinone compound, which exhibits anticancer effects on certain types of cancer cells. However, the effects of chrysophanol on human breast cancer remain to be elucidated. The aim of the present study was to clarify the role of chrysophanol on breast cancer cell lines MCF­7 and MDA­MB­231, and to identify the signal transduction pathways regulated by chrysophanol. MTT assay and flow cytometric analysis demonstrated that chrysophanol inhibited cell proliferation, and cell cycle progression in a dose­dependent manner. The expression of cell cycle­associated cyclin D1 and cyclin E were downregulated while p27 expression was upregulated following chrysophanol treatment at the mRNA, and protein levels. The Annexin V/propidium iodide staining assay results revealed that apoptosis levels increased following chrysophanol treatment. Chrysophanol upregulated caspase 3 and poly (ADP­ribose) polymerase cleavage in both cell lines. Furthermore, chrysophanol enhanced the effect of paclitaxel on breast cancer cell apoptosis. In addition, chrysophanol downregulated apoptosis regulator Bcl­2 protein, and transcription factor p65 and IκB phosphorylation. Inhbition of nuclear factor (NF)­κB by ammonium pyrrolidine dithiocarbamate diminished the effect of chrysophanol on apoptosis and associated proteins. In conclusion, the results of the current study demonstrated that chrysophanol effectively suppresses breast cancer cell proliferation and facilitates chemosentivity through modulation of the NF-κB signaling pathway.


Assuntos
Antraquinonas/farmacologia , Antineoplásicos/farmacologia , Ciclina D1/genética , Regulação Neoplásica da Expressão Gênica , Proteínas I-kappa B/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , Protocolos de Quimioterapia Combinada Antineoplásica , Apoptose/efeitos dos fármacos , Caspase 3/genética , Caspase 3/metabolismo , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ciclina D1/metabolismo , Ciclina E/genética , Ciclina E/metabolismo , Inibidor de Quinase Dependente de Ciclina p27/genética , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Humanos , Proteínas I-kappa B/metabolismo , Células MCF-7 , Paclitaxel/farmacologia , Poli(ADP-Ribose) Polimerases/genética , Poli(ADP-Ribose) Polimerases/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Transdução de Sinais , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/metabolismo
14.
Cancer Epidemiol ; 53: 21-26, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29353152

RESUMO

BACKGROUND: Gastric cancer is worldwide the third major cause of cancer related death. Risk factors for gastric cancer includes Helicobacter pylori infection, gastric ulcer, less hygienic condition, use of tobacco, alcohol consumption, use of salted, smoked food, genetic alterations etc. In order to identify the risk factors associated with gastric cancer in South Indian population a case-control study involving 200 proven gastric cancer cases and 400 controls was conducted. METHODS: A structured questionnaire was used to interview all the subjects who participated in our study. Genotyping assay was performed using Taqman allelic discrimination assay for 5 Single Nucleotide Polymorphisms (SNPs)-TGFß C-509T, TGFß T869C, XRCC1 Arg194Trp, IkBα C642T and IL4C-590T. RESULTS: Odds Ratios (ORs) and 95% confidence intervals (CIs) were calculated using conditional logistic regression. Statistical analysis on socio-economic factors, lifestyle factors had showed that subjects from low socio economic status, use of tobacco and consumption of non-vegetarian food had increased risk of developing gastric cancer. Multi-factorial analysis for the SNPs adjusting for the risk factors obtained in this study showed that TGFΒ C-509T TT genotypes had four fold increased risk of gastric cancer (OR = 4.11, CI = 1.02-16.56) and TGFß T869C CC genotype had a decreased risk of gastric cancer (OR = 0.21, CI = 0.05-0.85). CONCLUSION: Economic status, tobacco use and food habits play a significant role in gastric cancer development. TT genotype for TGFß C-509T had an increased risk and CC genotype for TGFß T869C had a decreased risk of gastric cancer in south Indian population after adjusting for socio-economic factors and lifestyle factors.


Assuntos
Predisposição Genética para Doença/genética , Estilo de Vida , Fatores Socioeconômicos , Neoplasias Gástricas/etiologia , Fator de Crescimento Transformador beta/genética , Alelos , Grupo com Ancestrais do Continente Asiático/genética , Estudos de Casos e Controles , Dieta , Feminino , Genótipo , Humanos , Proteínas I-kappa B/genética , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Polimorfismo de Nucleotídeo Único , Fatores de Risco , Proteína 1 Complementadora Cruzada de Reparo de Raio-X/genética
15.
J Biol Chem ; 293(7): 2452-2465, 2018 02 16.
Artigo em Inglês | MEDLINE | ID: mdl-29279332

RESUMO

Nuclear factor-κB (NF-κB) is a family of transcription factors that play a key role in cell survival and proliferation in many hematological malignancies, including multiple myeloma (MM). Bortezomib, a proteasome inhibitor used in the management of MM, can inhibit both canonical and noncanonical activation of NF-κB in MM cells. However, we previously reported that a significant fraction of freshly isolated MM cells harbor bortezomib-resistant NF-κB activity. Here, we report that hyaluronan and proteoglycan link protein 1 (HAPLN1) is produced in bone marrow stromal cells from MM patients, is detected in patients' bone marrow plasma, and can activate an atypical bortezomib-resistant NF-κB pathway in MM cells. We found that this pathway involves bortezomib-resistant degradation of the inhibitor of NF-κB (IκBα), despite efficient bortezomib-mediated inhibition of proteasome activity. Moreover, HAPLN1 can also confer bortezomib-resistant survival of MM cells. We propose that HAPLN1 is a novel pathogenic factor in MM that induces an atypical NF-κB activation and thereby promotes bortezomib resistance in MM cells.


Assuntos
Antineoplásicos/farmacologia , Bortezomib/farmacologia , Proteínas da Matriz Extracelular/metabolismo , Mieloma Múltiplo/metabolismo , NF-kappa B/metabolismo , Proteoglicanas/metabolismo , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Resistencia a Medicamentos Antineoplásicos , Proteínas da Matriz Extracelular/genética , Humanos , Proteínas I-kappa B/genética , Proteínas I-kappa B/metabolismo , Mieloma Múltiplo/tratamento farmacológico , Mieloma Múltiplo/genética , NF-kappa B/genética , Complexo de Endopeptidases do Proteassoma/genética , Complexo de Endopeptidases do Proteassoma/metabolismo , Proteoglicanas/genética , Proteólise
16.
Int J Occup Med Environ Health ; 31(4): 503-515, 2018 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-29165430

RESUMO

OBJECTIVES: To investigate the influence of the anti-tumor necrosis factor-α infliximab (IFX) in the case of rats with silicosis. MATERIAL AND METHODS: Forty-eight Wistar rats were randomly divided into 3 groups. The study group (N = 16) - silicosis was induced by intratracheal instillation of 50 mg silica on day 1, and IFX was subcutaneously administered at a dose of 15 mg/kg of body weight from day 2 to day 6, the vehicle group (N = 16) - silica used as the study group but without IFX, the sham group (N = 16) - 1 ml of saline was intratracheal-used. Eight rats in each group were euthanized on day 7 and on day 14, respectively. Lung tissue sections were stained with hematoxylin and eosin or Masson's trichromedye. The nuclear factor-κB p65 (NF-κB p65) positioning in the lung tissues were determined by immunohistochemical staining. Levels of tumor necrosis factor α (TNF-α) in rat serum and bronchoalveolar lavage fluid were measured with enzyme linked immunosorbent assay. The inducible nitric oxide synthase (iNOS) mRNA in the lung tissues was measured by quantitative real-time polymerase chain reaction, as well as inhibitor protein-κB (I-κB) and NF-κB p65 expression were measured quantitatively by western blotting. RESULTS: Silica installation increased the lung tissues inflammation reaction, oxidative stress and pulmonary fibrosis. Infliximab treatment significantly improved silica-induced lung pathological changes (inflammatory cells, collagen deposition), decreased the TNF-α inhibited NF-κB signaling (I-κB, NF-κB p65) as well as oxidant status (iNOS). CONCLUSIONS: Infliximab may improve silica-induced pulmonary inflammation by decreasing the TNF-α, inhibiting NF-κB signaling (I-κB, NF-κB p65) as well as oxidant status (iNOS), which suggest that IFX has potential role in the treatment of silica-induced lung damage. Int J Occup Med Environ Health 2018;31(4):503-515.


Assuntos
Infliximab/farmacologia , Fibrose Pulmonar/prevenção & controle , Dióxido de Silício , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Animais , Líquido da Lavagem Broncoalveolar/química , Proteínas I-kappa B/genética , Proteínas I-kappa B/metabolismo , Infliximab/administração & dosagem , Injeções Subcutâneas , Pulmão/patologia , Masculino , NF-kappa B , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Fibrose Pulmonar/induzido quimicamente , Ratos Wistar , Transdução de Sinais , Silicose/prevenção & controle , Fator de Necrose Tumoral alfa/sangue
17.
Drug Discov Ther ; 11(4): 206-211, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28867753

RESUMO

We have reported that 3a,4,5,9b-tetrahydro-4-(1-naphthalenyl)-3H-cyclopentan[c]quinoline-8-sulfonamide (TQS), α7 nicotinic acetylcholine receptor (nAChR) positive allosteric modulator (PAM) reduces lipopolysaccharide (LPS)-induced hyperalgesia and allodynia in mice. The objective of the present study was to determine the effects of TQS on LPS-induced activation of hippocampal inhibitor of κB (IκB) and cluster of differentiation 11b (CD11b) gene expression involving hyperalgesia and allodynia in mice. We also examined the effects of TQS on microglial phenotype following LPS administration. Pretreatment of TQS (4 mg/kg) reduced the expressions of IκB and CD11b mRNA. Pretreatment of methyllycaconitine (3 mg/kg), an α7 nAChR antagonist, reversed TQS-induced decrease in IκB and CD11b mRNA expressions in the hippocampus indicating the involvement of α7 nAChR. In addition, TQS (4 mg/kg) reversed the LPS-induced microglial morphological changes. These results suggest that TQS reduces LPS-induced IκB and CD11b gene expression and microglial activation associated with hyperalgesia and allodynia by targeting microglial α7 nAChR in the hippocampus.


Assuntos
Antígeno CD11b/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Proteínas I-kappa B/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Microglia/efeitos dos fármacos , Naftalenos/farmacologia , Quinolinas/farmacologia , Sulfonamidas/farmacologia , Receptor Nicotínico de Acetilcolina alfa7/efeitos dos fármacos , Aconitina/análogos & derivados , Aconitina/farmacologia , Regulação Alostérica , Animais , Antígeno CD11b/genética , Expressão Gênica/efeitos dos fármacos , Hipocampo/metabolismo , Proteínas I-kappa B/genética , Masculino , Camundongos , Microglia/patologia , Antagonistas Nicotínicos/farmacologia , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Receptor Nicotínico de Acetilcolina alfa7/antagonistas & inibidores
18.
Int J Mol Sci ; 18(8)2017 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-28933726

RESUMO

The NF-κB signaling pathway plays a pivotal role in regulating the immune response and inflammation. However, it has been shown that NF-κB also has a major role in oncogenesis. Therefore, NF-κB inhibitors have been considered as potential drugs against cancer. Herein, we searched for NF-κB inhibitors from natural sources and identified mollugin from the roots of Rubia cordifolia L. as an inhibitor of NF-κB activation. We found that mollugin significantly inhibited the expression of an NF-κB reporter gene induced by tumor necrosis factor (TNF)-α in a dose-dependent manner. Moreover, mollugin inhibited TNF-α-induced phosphorylation and nuclear translocation of p65, phosphorylation and degradation of inhibitor of κB (IκBα), and IκB kinase (IKK) phosphorylation. Furthermore, we discovered that pretreatment of cells with mollugin prevented the TNF-α-induced expression of NF-κB target genes, such as genes related to proliferation (COX-2, Cyclin D1 and c-Myc), anti-apoptosis (Bcl-2, cIAP-1 and survivin), invasion (MMP-9 and ICAM-1), and angiogenesis (VEGF). We also demonstrated that mollugin potentiated TNF-α-induced apoptosis and inhibited proliferation of HeLa cells. We further demonstrated in vivo that mollugin suppressed the growth of tumor xenografts derived from HeLa cells. Taken together, mollugin may be a valuable candidate for cancer treatment by targeting NF-κB.


Assuntos
NF-kappa B/genética , Neoplasias/tratamento farmacológico , Piranos/administração & dosagem , Fator de Necrose Tumoral alfa/genética , Animais , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Células HeLa , Humanos , Proteínas I-kappa B/genética , Camundongos , Inibidor de NF-kappaB alfa/genética , Proteínas de Neoplasias/genética , Neoplasias/genética , Neoplasias/patologia , Piranos/química , Fator de Transcrição RelA/genética , Ensaios Antitumorais Modelo de Xenoenxerto
19.
Microb Pathog ; 111: 440-445, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28923602

RESUMO

Candida albicans (C. albicans) is a commensal organism in human and a well-known dimorphic opportunistic pathogenic fungus. Though plenty of researches on the pathogenesis of C. albicans have been performed, the mechanism is not fully understood. The cell wall components of C. albicans have been documented to play important roles in its pathogenic processes. To further study the infectious mechanism of C. albicans, we investigated the potential functional role of its cell wall mannoprotein in cell cycle and apoptosis of HaCaT cells. We found that mannoprotein could promote the transition of cell cycle from G1/G0 to S phase, in which Cyclin D1, CDK4 and p-Rb, the major regulators of the cell cycle progression, showed significant upregulation, and CDKN1A (cyclin dependent kinase inhibitor 1A (p21)) showed significant downregulation. Mannoprotein also could inhibit apoptosis of HaCaT cells, which was well associated with increased expression of BCL2 (Bcl-2). Moreover, mannoprotein could increase the phosphorylation levels of RELA (p65) and NFKBIA (IκBα), as the key factors of NF-κB signal pathway in HaCaT cells, suggesting the activation of NF-κB signal pathway. Additionally, a NF-κB specific inhibitor, PDTC, could rescue the effect of mannoprotein on cell cycle and apoptosis of HaCaT cells, which suggested that mannoprotein could activate NF-κB signal pathway to mediate cell cycle alternation and inhibit apoptosis.


Assuntos
Apoptose , Candida albicans/metabolismo , Candidíase/metabolismo , Candidíase/microbiologia , Ciclo Celular , Parede Celular/metabolismo , Proteínas Fúngicas/metabolismo , Glicoproteínas de Membrana/metabolismo , NF-kappa B/metabolismo , Candida albicans/genética , Candidíase/genética , Candidíase/fisiopatologia , Linhagem Celular Tumoral , Parede Celular/genética , Ciclina D1/genética , Ciclina D1/metabolismo , Proteínas Fúngicas/genética , Interações Hospedeiro-Patógeno , Humanos , Proteínas I-kappa B/genética , Proteínas I-kappa B/metabolismo , Glicoproteínas de Membrana/genética , NF-kappa B/genética
20.
Haematologica ; 102(11): 1901-1912, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28775123

RESUMO

Chronic lymphocytic leukemia cells strongly depend on external stimuli for their survival. Both antigen receptor and co-stimulatory receptors, including Toll-like receptors, can modulate viability and proliferation of leukemic cells. Toll-like receptor ligands, and particularly the TLR9 ligand CpG, mediate heterogeneous responses in patients' samples reflecting the clinical course of the subjects. However, the molecular framework of the key signaling events underlying such heterogeneity is undefined. We focused our studies on a subset of chronic lymphocytic leukemia cases characterized by expression of CD38 and unmutated immunoglobulin genes, who respond to CpG with enhanced metabolic cell activity. We report that, while CpG induces NFKBIZ mRNA in all the samples analyzed, it induces the IκBζ protein in a selected group of cases, through an unanticipated post-transcriptional mechanism. Interestingly, IκBζ plays a causal role in sustaining CpG-induced cell viability and chemoresistance, and CpG stimulation can unleash immunoglobulin secretion by IκBζ-positive malignant cells. These results identify and characterize IκBζ as a marker and effector molecule of distinct key pathways in chronic lymphocytic leukemia.


Assuntos
Regulação Leucêmica da Expressão Gênica , Proteínas I-kappa B/genética , Imunoglobulina M/biossíntese , Leucemia Linfocítica Crônica de Células B/genética , Leucemia Linfocítica Crônica de Células B/metabolismo , Proteínas Nucleares/genética , Receptor Toll-Like 9/agonistas , Autofagia , Biomarcadores , Células Cultivadas , Regulação Leucêmica da Expressão Gênica/efeitos dos fármacos , Humanos , Imunofenotipagem , Leucemia Linfocítica Crônica de Células B/patologia , Oligodesoxirribonucleotídeos/farmacologia , Processamento Pós-Transcricional do RNA
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