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1.
Mol Biol Rep ; 2020 Dec 08.
Artigo em Inglês | MEDLINE | ID: mdl-33289908

RESUMO

Porphyromonas gingivalis triggers a range of innate immune responses in the host that may contribute to the development of periodontitis and dementing diseases including Alzheimer's disease (AD). This study aimed to assess the mode of action of trans-resveratrol in modulating the P. gingivalis lipopolysaccharide (PgLPS) induced metabolic inflammation in a neuronal cell model. Confluent IMR-32 neuroblastoma cells were treated with trans-resveratrol from Polygonum cuspidatum in the presence or absence of PgLPS. The abundance of messenger ribo-nucleic acid (mRNA) transcripts of a panel of 92 genes was quantitatively assessed through targeted transcriptome profiling technique and the biochemical pathways affected were identified through Ingenuity Pathway Analysis. Gene expression analysis revealed that trans-resveratrol down-regulated the mRNA of multiple gene markers including growth factors, transcription factors, kinases, trans-membrane receptors, cytokines and enzymes that were otherwise activated by PgLPS treatment of IMR-32 neuroblastoma cells. Pathway analysis demonstrated that the cellular oxidative stress caused by the activation of phosphoinositide-3-kinase/Akt1 (PI3K/Akt1) pathway that leads to the production of reactive oxygen species (ROS), chronic inflammatory response induced by the activation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) pathway and nutrient utilization pathways were favourably modulated by trans-resveratrol in the PgLPS challenged IMR-32 cells. This study demonstrates the potential of trans-resveratrol as a bioactive compound with multiple modes of intracellular action further supporting its therapeutic application in neuroinflammatory diseases.

2.
Vet Ital ; 56(3): 185-192, 2020 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-33543914

RESUMO

Sumac (Rhus coriaria L.) is a plant species belong to Anacardiaceous family that is worldwide diffused. The sumac seed power (SSP), produced by grinding dried fruits, is recognized to have defensive and beneficial effects on numerous health­related problems. In this study, SSP was included in broilers basal­diet to investigate the comparative effects of different levels of SSP on performance, carcass characteristics, blood parameters, immune system and ileal microorganisms. A total of 225, one day­old male broilers (Ross 308) were randomly assigned to the five dietary treatments with three replicates per treatment. The experimental diets were: basal­diet (BD); and BD including 0.05, 0.10, 0.15 and 0.20% SSP, respectively. During the whole feeding period (42 days), birds fed corn­based grower (1­21 days) and finisher (22­42 days) diets, respectively. Results indicated that supplementing SSP had no effect on broiler body weight gain, feed intake and feed conversion as well as carcass characteristics (P > 0.05). Similarly, blood total protein, albumin, glucose and triglyceride were not influenced by dietary SSP. Conversely, serum total cholesterol and LDL­cholesterol levels were decreased, while HDL­cholesterol increased in all SSP fed groups compared to control (P < 0.05). In this study the addition of SSP in broilers diets did not show any effect on blood heterophils and lymphocyte. Moreover, the lactobacillus count remained unaffected by dietary treatments, while E. coli count in broiler ileal content was lower when fed 0.10% SSP than the other groups (P < 0.05). Thus, the present findings indicated a positive effect of feeding SSP (especially at 0.10% diet) on blood cholesterol levels and E. coli count in broiler chickens

3.
Curr Pharm Des ; 24(5): 595-614, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29278208

RESUMO

BACKGROUND: Potassium bromate (KBrO3), a food additive, has been used in many bakery products as an oxidizing agent. It has been shown to induce renal cancer in many in-vitro and in-vivo experimental models. OBJECTIVES: This study evaluated the carcinogenic potential of potassium bromate (KBrO3) and the chemopreventive mechanisms of the anti-oxidant and anti-inflammatory phytochemical, curcumin against KBrO3-induced carcinogenicity. METHOD: Lactate dehydrogenase (LDH) cytotoxicity assay and morphological characteristics were used to assess curcumin's cytoprotective potential against KBrO3 toxicity. To assess the chemopreventive potential of curcumin against KBrO3-induced oxidative insult, intracellular H2O2 and the nuclear concentration of the DNA adduct 8- OHdG were measured. PCR array, qRT-PCR, and western blot analysis were used to identify dysregulated genes by KBrO3 exposure. Furthermore, immunofluorescence was used to evaluate the ciliary loss and the disturbance of cellular tight junction induced by KBrO3. RESULTS: Oxidative stress assays showed that KBrO3 increased the levels of intracellular H2O2 and the DNA adduct 8-OHdG. Combination of curcumin with KBrO3 efficiently reduced the level of H2O2 and 8-OHdG while upregulating the expression of catalase. PCR array, qRT-PCR, and western blot analysis revealed that KBrO3 dysregulated multiple genes involved in inflammation, proliferation, and apoptosis, namely CTGF, IL-1, and TRAF3. Moreover, qRT-PCR and immunofluorescence studies showed that KBrO3 negatively affected the tight junctional protein (ZO-1) and induced a degeneration of primary ciliary proteins. The negative impact of KBrO3 on cilia was markedly repressed by curcumin. CONCLUSION: Curcumin could potentially be used as a protective agent against carcinogenicity of KBrO3.


Assuntos
Bromatos/antagonistas & inibidores , Carcinógenos/antagonistas & inibidores , Curcumina/farmacologia , Aditivos Alimentares/efeitos adversos , Substâncias Protetoras/farmacologia , Bromatos/farmacologia , Carcinógenos/farmacologia , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Curcumina/análise , Relação Dose-Resposta a Droga , Células Epiteliais/efeitos dos fármacos , Humanos , Substâncias Protetoras/análise
4.
Front Immunol ; 8: 7, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28167941

RESUMO

Dysregulation of inflammatory responses is a hallmark of multiple diseases such as atherosclerosis and rheumatoid arthritis. As constitutively active transcription factors, NR4A nuclear receptors function to control the magnitude of inflammatory responses and in chronic inflammatory disease can be protective or pathogenic. Within this study, we demonstrate that TLR4 stimulation using the endotoxin lipopolysaccharide (LPS) rapidly enhances NR4A1-3 expression in human and murine, primary and immortalized myeloid cells with concomitant gene transcription and protein secretion of MIP-3α, a central chemokine implicated in numerous pathologies. Deficiency of NR4A2 and NR4A3 in human and murine myeloid cells reveals that both receptors function as positive regulators of enhanced MIP-3α expression. In contrast, within the same cell types and conditions, altered NR4A activity leads to suppression of LPS-induced MCP-1 gene and protein expression. An equivalent pattern of inflammatory gene regulation is replicated in TNFα-treated myeloid cells. We show that NF-κB is the critical regulator of NR4A1-3, MIP-3α, and MCP-1 during TLR4 stimulation in myeloid cells and highlight a parallel mechanism whereby NR4A activity can repress or enhance NF-κB target gene expression simultaneously. Mechanistic insight reveals that NR4A2 does not require DNA-binding capacity in order to enhance or repress NF-κB target gene expression simultaneously and establishes a role for NF-κB family member Relb as a novel NR4A target gene involved in the positive regulation of MIP-3α. Thus, our data reveal a dynamic role for NR4A receptors concurrently enhancing and repressing NF-κB activity in myeloid cells leading to altered transcription of key inflammatory mediators.

5.
J Innate Immun ; 9(2): 203-216, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-27902980

RESUMO

BACKGROUND: Chronic inflammation and oxidative stress are hallmarks of chagasic cardiomyopathy (CCM). In this study, we determined if microparticles (MPs) generated during Trypanosoma cruzi (Tc) infection carry the host's signature of the inflammatory/oxidative state and provide information regarding the progression of clinical disease. METHODS: MPs were harvested from supernatants of human peripheral blood mononuclear cells in vitro incubated with Tc (control: LPS treated), plasma of seropositive humans with a clinically asymptomatic (CA) or symptomatic (CS) disease state (vs. normal/healthy [NH] controls), and plasma of mice immunized with a protective vaccine before challenge infection (control: unvaccinated/infected). Macrophages (mφs) were incubated with MPs, and we probed the gene expression profile using the inflammatory signaling cascade and cytokine/chemokine arrays, phenotypic markers of mφ activation by flow cytometry, cytokine profile by means of an ELISA and Bioplex assay, and oxidative/nitrosative stress and mitotoxicity by means of colorimetric and fluorometric assays. RESULTS: Tc- and LPS-induced MPs stimulated proliferation, inflammatory gene expression profile, and nitric oxide (∙NO) release in human THP-1 mφs. LPS-MPs were more immunostimulatory than Tc-MPs. Endothelial cells, T lymphocytes, and mφs were the major source of MPs shed in the plasma of chagasic humans and experimentally infected mice. The CS and CA (vs. NH) MPs elicited >2-fold increase in NO and mitochondrial oxidative stress in THP-1 mφs; however, CS (vs. CA) MPs elicited a more pronounced and disease-state-specific inflammatory gene expression profile (IKBKB, NR3C1, and TIRAP vs. CCR4, EGR2, and CCL3), cytokine release (IL-2 + IFN-γ > GCSF), and surface markers of mφ activation (CD14 and CD16). The circulatory MPs of nonvaccinated/infected mice induced 7.5-fold and 40% increases in ∙NO and IFN-γ production, respectively, while these responses were abolished when RAW264.7 mφs were incubated with circulatory MPs of vaccinated/infected mice. CONCLUSION: Circulating MPs reflect in vivo levels of an oxidative, nitrosative, and inflammatory state, and have potential utility in evaluating disease severity and the efficacy of vaccines and drug therapies against CCM.


Assuntos
Micropartículas Derivadas de Células/metabolismo , Doença de Chagas/imunologia , Inflamação/imunologia , Leucócitos Mononucleares/imunologia , Macrófagos/imunologia , Trypanosoma cruzi/imunologia , Vacinas/imunologia , Animais , Doenças Assintomáticas , Linhagem Celular , Micropartículas Derivadas de Células/imunologia , Citocinas/metabolismo , Progressão da Doença , Feminino , Perfilação da Expressão Gênica , Humanos , Inflamação/genética , Macrófagos/parasitologia , Camundongos , Camundongos Endogâmicos C57BL , Óxido Nítrico/metabolismo , Estresse Oxidativo , Vacinação
6.
Eur J Nutr ; 55(5): 1951-62, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26254196

RESUMO

PURPOSE: Palmaria palmata (P. Palmata) is reported to contain anti-inflammatory and antioxidant compounds albeit no study has investigated these effects in humans. METHODS: A randomised parallel placebo-controlled human intervention study was carried out to investigate the effect of consuming P. Palmata (5 g/day) incorporated into a bread on serum markers of inflammation [C-reactive protein (CRP); cytokine analysis] with secondary analysis investigating changes in lipids (cholesterol, triglycerides), thyroid function [thyroid-stimulating hormone (TSH)] and antioxidant status ferric reducing antioxidant power. ANCOVA with baseline values as covariates, controlling for age, BMI, sex and smoking status, was used to compare differences between treatment groups over time . In vitro studies investigated the inflammatory activity of P. Palmata extracts (hot water, cold water and ethanol extract), protein extracts and associated protein hydrolysates using a Caco-2 inflammation cell model. RESULTS: Consumption of P. Palmata-enriched bread significantly increased serum CRP (+16.1 %, P = 0.011), triglycerides (+31.9 %, P = 0.001) and TSH (+17.2 %, P = 0.017) when compared to the control group. In vitro evaluation of P. palmata extracts and protein hydrolysates identified a significant induction of IL-8 secretion by Caco-2 cells, and the hot water P. palmata extract was shown to increase adipocyte glycerol release (P < 0.05). CONCLUSION: Evidence from this human study suggests that P. palmata stimulates inflammation, increases serum triglycerides and alters thyroid function; however, these changes are not likely to impact health as changes remained within the normal clinical range. The data from the in vitro study provided indications that IL-8 may contribute to the apparent immunostimulation noted in the human study.


Assuntos
Pão/análise , HDL-Colesterol/sangue , LDL-Colesterol/sangue , Rodófitas/química , Glândula Tireoide/metabolismo , Triglicerídeos/sangue , Células 3T3-L1 , Adipócitos , Adolescente , Adulto , Idoso , Animais , Antioxidantes/metabolismo , Biomarcadores/sangue , Índice de Massa Corporal , Células CACO-2 , Dieta , Método Duplo-Cego , Feminino , Humanos , Interferon gama/sangue , Interleucinas/sangue , Masculino , Camundongos , Pessoa de Meia-Idade , Estresse Oxidativo , Extratos Vegetais/análise , Proteínas de Plantas/análise , Alga Marinha/química , Fator de Necrose Tumoral alfa/sangue , Adulto Jovem
7.
Mar Drugs ; 13(8): 5402-24, 2015 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-26308008

RESUMO

Algae contain a number of anti-inflammatory bioactive compounds such as omega-3 polyunsaturated fatty acids (n-3 PUFA) and chlorophyll a, hence as dietary ingredients, their extracts may be effective in chronic inflammation-linked metabolic diseases such as cardiovascular disease. In this study, anti-inflammatory potential of lipid extracts from three red seaweeds (Porphyra dioica, Palmaria palmata and Chondrus crispus) and one microalga (Pavlova lutheri) were assessed in lipopolysaccharide (LPS)-stimulated human THP-1 macrophages. Extracts contained 34%-42% total fatty acids as n-3 PUFA and 5%-7% crude extract as pigments, including chlorophyll a, ß-carotene and fucoxanthin. Pretreatment of the THP-1 cells with lipid extract from P. palmata inhibited production of the pro-inflammatory cytokines interleukin (IL)-6 (p < 0.05) and IL-8 (p < 0.05) while that of P. lutheri inhibited IL-6 (p < 0.01) production. Quantitative gene expression analysis of a panel of 92 genes linked to inflammatory signaling pathway revealed down-regulation of the expression of 14 pro-inflammatory genes (TLR1, TLR2, TLR4, TLR8, TRAF5, TRAF6, TNFSF18, IL6R, IL23, CCR1, CCR4, CCL17, STAT3, MAP3K1) by the lipid extracts. The lipid extracts effectively inhibited the LPS-induced pro-inflammatory signaling pathways mediated via toll-like receptors, chemokines and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling molecules. These results suggest that lipid extracts from P. lutheri, P. palmata, P. dioica and C. crispus can inhibit LPS-induced inflammatory pathways in human macrophages. Therefore, algal lipid extracts should be further explored as anti-inflammatory ingredients for chronic inflammation-linked metabolic diseases.


Assuntos
Anti-Inflamatórios/farmacologia , Lipídeos/química , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Rodófitas/metabolismo , Células Cultivadas , Regulação para Baixo/efeitos dos fármacos , Humanos , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Macrófagos/metabolismo , Microalgas/efeitos dos fármacos , Microalgas/metabolismo , NF-kappa B/metabolismo , Alga Marinha/metabolismo , Transdução de Sinais/efeitos dos fármacos , Receptores Toll-Like/metabolismo
8.
J Immunol ; 195(4): 1436-48, 2015 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-26150530

RESUMO

Adenosine receptor-mediated regulation of monocyte/macrophage inflammatory responses is critical in the maintenance of tissue homeostasis. In this study, we reveal that adenosine potently modulates the expression of NR4A1, 2, and 3 orphan nuclear receptors in myeloid cells, and this modulation is primarily through the adenosine A2a receptor subtype. We demonstrate that A2a receptor activation of NR4A1-3 receptor synthesis is further enhanced in TLR4-stimulated monocytes. After TLR4 stimulation, NR4A receptor-depleted monocyte/macrophage cells display significantly altered expression of cell-surface markers and produce increased inflammatory cytokine and chemokine secretion rendering the cells an enhanced proinflammatory phenotype. Exposure of TLR4 or TNF-α-stimulated monocytes to adenosine analogs directs changes in the expression of MIP-3α and IL-23p19, with NR4A2 depletion leading to significantly enhanced expression of these factors. Furthermore, we establish that nuclear levels of NF-κB/p65 are increased in TLR/adenosine-stimulated NR4A2-depleted cells. We show that, after TLR/adenosine receptor stimulation, NR4A2 depletion promotes significant binding of NF-κB/p65 to a κB consensus binding motif within the MIP-3α proximal promoter leading to increased protein secretion, confirming a pivotal role for NF-κB activity in controlling cellular responses and gene expression outcomes in response to these mediators. Thus, these data demonstrate that during an inflammatory response, adenosine modulation of NR4A receptor activity acts to limit NF-κB-mediated effects and that loss of NR4A2 expression leads to enhanced NF-κB activity and hyperinflammatory responses in myeloid cells.


Assuntos
Adenosina/metabolismo , Monócitos/imunologia , Monócitos/metabolismo , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/metabolismo , Animais , Linhagem Celular , Quimiocina CCL20/genética , Proteínas de Ligação a DNA/genética , Regulação da Expressão Gênica , Humanos , Hipersensibilidade/genética , Hipersensibilidade/imunologia , Hipersensibilidade/metabolismo , Inflamação/genética , Inflamação/imunologia , Inflamação/metabolismo , Subunidade p19 da Interleucina-23/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos , Modelos Biológicos , NF-kappa B/metabolismo , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/genética , Membro 2 do Grupo A da Subfamília 4 de Receptores Nucleares/genética , Membro 2 do Grupo A da Subfamília 4 de Receptores Nucleares/metabolismo , Membro 3 do Grupo A da Subfamília 4 de Receptores Nucleares/genética , Membro 3 do Grupo A da Subfamília 4 de Receptores Nucleares/metabolismo , Receptores Nucleares Órfãos , Receptor A2A de Adenosina/metabolismo
9.
Eur J Pharm Biopharm ; 94: 194-206, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26026287

RESUMO

Epithelial damage caused by intestinal permeation enhancers is a source of debate concerning safety. The medium chain fatty acid, sodium caprate (C10), causes reversible membrane perturbation at high dose levels required for efficacy in vivo, so the aim was to model it in vitro. Exposure of Caco-2 monolayers to 8.5mM C10 for 60min followed by incubation in fresh buffer led to (i) recovery in epithelial permeability (i.e. transepithelial electrical resistance (TEER) and apparent permeability coefficient (Papp) of [(14)C]-mannitol), (ii) recovery of cell viability parameters (monolayer morphology, plasma membrane potential, mitochondrial membrane potential, and intracellular calcium) and (iii) reduction in mRNA expression associated with inflammation (IL-8). Pre-incubation of monolayers with a mucosal prostaglandin cytoprotectant was attempted in order to further decipher the mechanism of C10. Misoprostol (100nM), inhibited C10-induced changes in monolayer parameters, an effect that was partially attenuated by the EP1 receptor antagonist, SC51322. In rat isolated intestinal tissue mucosae and in situ loop instillations, C10-induced respective increases in the [(14)C]-mannitol Papp and the AUC of FITC-dextran 4000 (FD-4) were similarly inhibited by misoprostol, with accompanying morphological damage spared. These data support a temporary membrane perturbation effect of C10, which is linked to its capacity to mainly increase paracellular flux, but which can be prevented by pre-exposure to misoprostol.


Assuntos
Colo/efeitos dos fármacos , Ácidos Decanoicos/toxicidade , Células Epiteliais/efeitos dos fármacos , Absorção Intestinal/efeitos dos fármacos , Mucosa Intestinal/efeitos dos fármacos , Misoprostol/farmacologia , Substâncias Protetoras/farmacologia , Animais , Células CACO-2 , Sobrevivência Celular/efeitos dos fármacos , Colo/metabolismo , Colo/patologia , Citoproteção , Dextranos/metabolismo , Relação Dose-Resposta a Droga , Impedância Elétrica , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/metabolismo , Regulação da Expressão Gênica , Humanos , Técnicas In Vitro , Interleucina-8/genética , Interleucina-8/metabolismo , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Manitol/metabolismo , Permeabilidade , RNA Mensageiro/metabolismo , Ratos , Fatores de Tempo
10.
PLoS One ; 10(5): e0124823, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25978040

RESUMO

Texel lambs are known to be more resistant to gastrointestinal nematode (GIN) infection than Suffolk lambs, with a greater ability to limit infection. The objectives of this study were to: 1) profile the whole transcriptome of abomasal lymph node tissue of GIN-free Texel and Suffolk lambs; 2) identify differentially expressed genes and characterize the immune-related biological pathways and networks associated with these genes. Abomasal lymph nodes were collected from Texel (n = 6) and Suffolk (n = 4) lambs aged 19 weeks that had been GIN-free since 6 weeks of age. Whole transcriptome profiling was performed using RNA-seq on the Illumina platform. At the time of conducting this study, a well annotated Ovine genome was not available and hence the sequence reads were aligned with the Bovine (UMD3.1) genome. Identification of differentially expressed genes was followed by pathway and network analysis. The Suffolk breed accounted for significantly more of the differentially expressed genes, (276 more highly expressed in Suffolk v 162 in Texel; P < 0.001). The four most significant differentially expressed pathways were all related to immunity and were classified as: Role of Pattern Recognition Receptors in Recognition of Bacteria and Viruses, Activation of IRF by Cytosolic Pattern Recognition Receptors, Role of RIG-I-like Receptors in Antiviral Innate Immunity, and Interferon Signaling. Of significance is the fact that all of these four pathways were more highly expressed in the Suffolk. These data suggest that in a GIN-free environment, Suffolk lambs have a more active immune profile relative to the Texel: this immune profile may contribute to the poorer efficiency of response to a GIN challenge in the Suffolk breed compared to the Texel breed.


Assuntos
Linfonodos/metabolismo , Nematoides/patogenicidade , Infecções por Nematoides/genética , Infecções por Nematoides/parasitologia , Doenças dos Ovinos/genética , Animais , Bovinos , Ovinos , Doenças dos Ovinos/parasitologia , Transcriptoma/genética
11.
Food Sci Nutr ; 2(6): 712-23, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25493190

RESUMO

Bioactive milk peptides are reported to illicit a range of physiological benefits and have been proposed as potential functional food ingredients. The objective of this study was to characterize the anti-inflammatory properties of sodium caseinate (NaCAS), its enzyme hydrolysate (EH) and peptide-enriched fractions (5 kDa retentate [R], 1 kDaR and 1 kDa permeate [P]), both in vitro using a Caco-2 cell line, and also ex vivo using a porcine colonic tissue explant system. Caco-2 cells were stimulated with tumour necrosis factor alpha (TNFα) and co-treated with casein hydrolysates for 24 h. Following this, interleukin (IL)-8 concentrations in the supernatant were measured using enzyme-linked immunosorbent assay. Porcine colonic tissue was stimulated with lipopolysaccharide and co-treated with casein hydrolysates for 3 h. The expression of a panel of inflammatory cytokines was measured using qPCR. While dexamethasone reduced the IL-8 concentration by 41.6%, the 1 kDaR and 1 kDaP fractions reduced IL-8 by 68.7% and 66.1%, respectively, relative to TNFα-stimulated Caco-2 cells (P < 0.05). In the ex vivo system, only the 1 kDaR fraction elicited a decrease inIL1-α,IL1-ß,IL-8,TGF-ß andIL-10 expression (P < 0.05). This study provides evidence that the bioactive peptides present in the 1 kDaR fraction of the NaCAS hydrolysate possess anti-inflammatory properties in vitro and ex vivo. Further in vivo analysis of the anti-inflammatory properties of the 1 kDaR is proposed.

12.
Mol Immunol ; 62(1): 159-68, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25019566

RESUMO

Active pulmonary tuberculosis (APTB) is associated with a failure of the host immune system to control the invading Mycobacterium tuberculosis (Mtb). The objective of this study was to quantify and assess the role of serum inflammatory cytokines in active pulmonary tuberculosis patients following anti-tuberculosis drug (ATD) therapy. Blood samples were collected from APTB patients and normal healthy subjects (NHS) (total n=204) at baseline and 2, 4 and 6 months post-therapy and the abundance of serum inflammatory cytokines were measured by cytokine specific ELISA. Compared to NHS, APTB patients at baseline had higher levels of serum pro-inflammatory cytokines IL-12p40 (P<0.001), IFN-γ (P<0.001), TNF-α (P<0.01), IL-1ß (P<0.001) and IL-6 (P<0.001) and anti-inflammatory cytokines IL-10 (P<0.001) and TGF-ß1 (P<0.001) while there was no change in the level of IL-4. In APTB patients, the serum levels of IFN-γ, TNF-α, IL-6 and TGF-ß1 directly relate to the bacterial load while the TNF-α, IL-1ß, IL-6 and TGF-ß1 relate to radiological severity. At baseline, the IL-6 level in NHS and APTB patients differed most and following ATD therapy, this level rapidly decreased and stabilized by 4-month in APTB patients. It is concluded that a subtle reduction in the serum level of IL-6 of the APTB patients following ATD therapy might play a vital role in immune-protection of the host against Mtb infection and hence the serum IL-6 level can be a useful marker to diagnose the effectiveness of therapy in the patients.


Assuntos
Antituberculosos/uso terapêutico , Citocinas/sangue , Mediadores da Inflamação/sangue , Tuberculose Pulmonar/sangue , Tuberculose Pulmonar/tratamento farmacológico , Adulto , Estudos de Casos e Controles , Feminino , Humanos , Interferon gama/sangue , Subunidade p40 da Interleucina-12/sangue , Interleucina-6/sangue , Masculino , Prognóstico , Fator de Crescimento Transformador beta1/sangue , Resultado do Tratamento , Tuberculose Pulmonar/diagnóstico , Fator de Necrose Tumoral alfa/sangue
13.
Rapid Commun Mass Spectrom ; 28(9): 1011-8, 2014 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-24677522

RESUMO

RATIONALE: Isotope ratio analysis of bovine tissues is a tool for inferring aspects of the dietary history of cattle. The objective of this experiment was to quantify the carbon (C) and nitrogen (N) isotopic turnover in blood (serum and residue) and inner organs (liver, kidney, heart and brain) of beef cattle. METHODS: Growing beef cattle (n = 70 in total) were either switched from a control diet containing barley and urea to an experimental diet containing maize and (15)N-enriched urea, for various intervals prior to slaughter or maintained on the control diet for 168 days pre-slaughter. Samples of blood, liver, kidney, heart and brain were collected at 0, 14, 28, 56, 112 and 168 days and analysed using Isotope Ratio Mass Spectrometry. RESULTS: After 168 days, C- and N-isotopic equilibrium was reached in the blood serum, liver and kidney, approached in the heart and brain, but not reached in the non-serum component of blood. The estimated C and N half-lives were 16.5 and 20.7 days for liver, 19.2 and 25.5 days for kidney, 29.2 and 35.6 days for blood serum, 37.6 and 49.9 days for heart, 53.3 and 52.2 days for brain and 113.3 and 115.0 days for the non-serum blood residue, respectively. Modelling the C and N turnover in the different tissue combinations revealed that a combined analysis of liver and heart as well as brain and kidney can provide the most accurate estimation of the timing of the diet switch. CONCLUSIONS: Based on the difference in turnover rates, bovine soft tissues can provide isotopic information on short- and long-term dietary changes, which in turn may be linked to the geographic or production origin of beef cattle. This study also provides basic biological data on organ C and N turnover in a large herbivorous mammal.


Assuntos
Isótopos de Carbono/análise , Isótopos de Carbono/metabolismo , Isótopos de Nitrogênio/análise , Isótopos de Nitrogênio/metabolismo , Ração Animal , Animais , Química Encefálica , Isótopos de Carbono/administração & dosagem , Isótopos de Carbono/sangue , Bovinos , Rim/química , Fígado/química , Espectrometria de Massas/métodos , Isótopos de Nitrogênio/administração & dosagem , Isótopos de Nitrogênio/sangue , Distribuição Tecidual
14.
PLoS One ; 8(3): e60011, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23544120

RESUMO

Chito-oligosaccharide (COS) is a natural bioactive compound, which has been shown to suppress lipid metabolic genes and lipid accumulation in differentiating adipocytes. Leptin has been identified as a key regulator of energy homeostasis and is known to be under epigenetic regulation during adipogenesis. Hence, the first objective of this experiment was to compare leptin gene (LEP) expression and leptin secretion during the different stages of adipogenesis and to investigate the effect of COS on these processes. As COS inhibited LEP expression during adipogenesis, the second aim was to investigate the methylation dynamics of a 'CpG' island in the proximal region of the LEP promoter during adipogenesis and to determine the effect of COS on this process. Mouse 3T3-L1 cells were stimulated to differentiate in the absence or presence of COS and the levels of leptin mRNA and protein were evaluated on days 0, 2, 4 and 6 post-induction of differentiation (PID). The extent of de-methylation of six CpG sites was evaluated. LEP mRNA transcript and protein could not be detected on either day 0PID or 2PID. In contrast, both were detected on day 4PID (P<0.05) and 6PID (P<0.001) and both were inhibited by COS (P<0.001). Of the six CpG sites analyzed, CpG_52, CpG_62 and CpG_95 became 11.5, 5.0 and 5.0% de-methylated between day 2PID and 6PID, respectively. COS blocked this de-methylation event at CpG_52 (P<0.001), CpG_62 (P<0.01) and CpG_95 (P<0.01) on day 6PID. These data suggest that COS can have an epigenetic effect on differentiating adipocytes, a novel biological function of COS which has potential applications for the manipulation of leptin gene expression, adipogenesis, and conditions within the metabolic syndrome spectrum.


Assuntos
Adipogenia/genética , Quitosana/farmacologia , Ilhas de CpG/genética , Metilação de DNA/genética , Leptina/genética , Oligossacarídeos/farmacologia , Regiões Promotoras Genéticas , Células 3T3-L1 , Adipogenia/efeitos dos fármacos , Animais , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Metilação de DNA/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Leptina/metabolismo , Camundongos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transcrição Genética/efeitos dos fármacos
15.
PLoS One ; 8(1): e53828, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23342013

RESUMO

Chitosan, a natural polysaccharide comprising copolymers of glucosamine and N-acetylglucosamine, has been shown to have anti-obesity properties. Two experiments (Exp. 1 and Exp. 2) were performed to determine the role of chitosan on dietary intake, body weight gain, and fat deposition in a pig model, as well as identifying potential mechanisms underlying the anti-obesity effect of chitosan. In Exp. 1, the nutrient digestibility experiment, 16 pigs (n = 4/treatment) were randomly allocated to one of four dietary treatments as follows: 1) basal diet; 2) basal diet plus 300 ppm chitosan; 3) basal diet plus 600 ppm chitosan; 4) basal diet plus 1200 ppm chitosan. The main observation was that crude fat digestibility was lower in the 1200 ppm chitosan group when compared with the control group (P<0.05). In Exp. 2, a total of 80 pigs (n = 20/treatment) were offered identical dietary treatments to that offered to animals in Exp. 1. Blood samples were collected on day 0, day 35 and at the end of the experiment (day 57). Animals offered diets containing 1200 ppm chitosan had a lower daily dietary intake (P<0.001) and body weight gain (P<0.001) from day 35 to 57 when compared with all the other treatment groups. Animals offered diets containing 1200 ppm chitosan had a significantly lower final body weight (P<0.01) when compared with all the other treatment groups. The decreased dietary intake observed in the 1200 ppm chitosan group was associated with increased serum leptin concentrations (P<0.001) and a decrease in serum C-reactive protein (CRP) concentrations (P<0.05). In conclusion, the results of this study highlight novel endocrine mechanisms involving the modulation of serum leptin and CRP concentrations by which chitosan exhibits anti-obesity properties in vivo.


Assuntos
Fármacos Antiobesidade/farmacologia , Quitosana/farmacologia , Obesidade/prevenção & controle , Adipocinas/sangue , Animais , Peso Corporal/efeitos dos fármacos , Suplementos Nutricionais , Digestão/efeitos dos fármacos , Ingestão de Alimentos/efeitos dos fármacos , Obesidade/sangue , Obesidade/dietoterapia , Obesidade/fisiopatologia , Suínos
16.
Mol Immunol ; 51(3-4): 283-91, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22512945

RESUMO

Chitooligosaccharides (COSs) are bioactive carbohydrate derivatives that have numerous health benefits, including stimulation of the immune system. The objectives of this study were to evaluate the effect of chitooligosaccharide (COS) on expression of a specific panel of cytokine genes involved in inflammation and to delineate the signal transduction pathway underlying the COS mediated inflammatory response. Human intestinal epithelial-like (Caco-2) cells were treated with COS (5000-10,000Da) and expression of a panel of eighty-four cytokine genes was analyzed by quantitative real-time PCR. COS induced up-regulation of a total of 11 genes including CCL20 and IL8 and concurrent down-regulation of 10 genes including pro-inflammatory mediators CCL15, CCL25 and IL1B. To further establish the signal transduction pathway of COS mediated response in Caco-2 cells, two major inflammatory signal transduction pathways (NF-κB and AP-1) were investigated. COS had inhibitory effect (P<0.01) on TNF-α induced NF-κB binding activity while stimulatory effect (P<0.001) on AP-1 binding activity. COS also inhibited the expression of RELA (P<0.01) and IKBKB (P<0.01) genes of NF-κB pathway while stimulate the expression of JUN (P<0.05) gene of AP-1 pathway. In conclusion, COS elicits an acute inflammatory cytokine response in Caco-2 cells and hence it has the potential to stimulate the immune system in the gut epithelium.


Assuntos
Inflamação/metabolismo , Oligossacarídeos/farmacologia , Fator de Transcrição AP-1/metabolismo , Células CACO-2 , Células Cultivadas , Quimiocina CCL20/genética , Quimiocina CCL20/metabolismo , Quimiocinas CC/genética , Quimiocinas CC/metabolismo , Regulação para Baixo/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Humanos , Inflamação/induzido quimicamente , Inflamação/genética , Inflamação/patologia , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Interleucina-8/genética , Interleucina-8/metabolismo , Proteínas Inflamatórias de Macrófagos/genética , Proteínas Inflamatórias de Macrófagos/metabolismo , NF-kappa B/genética , NF-kappa B/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fator de Transcrição AP-1/genética , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Regulação para Cima/efeitos dos fármacos
17.
J Clin Microbiol ; 50(6): 2082-4, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22461679

RESUMO

Conventional and molecular techniques were applied to detect and characterize drug resistance of mycobacteria in the sputum samples of clinically confirmed tuberculosis. The sensitivities of mycobacterium detection by ZN staining, culture, multiplex PCR, and restriction fragment length polymorphism (RFLP) were 27.7%, 19.9%, 92.9%, and 95.7%, respectively, but all were 100% specific. The conventional and multiple-allele-specific PCR (MAS-PCR) methods enabled establishment of the drug resistance in 19.3% and 86.9% cases, respectively. We demonstrated that molecular techniques have potential in the accurate diagnosis of tuberculosis.


Assuntos
Antituberculosos/farmacologia , Farmacorresistência Bacteriana , Mycobacterium tuberculosis/isolamento & purificação , Escarro/microbiologia , Tuberculose Resistente a Múltiplos Medicamentos/diagnóstico , Tuberculose Pulmonar/diagnóstico , Humanos , Testes de Sensibilidade Microbiana/métodos , Reação em Cadeia da Polimerase Multiplex/métodos , Mycobacterium tuberculosis/efeitos dos fármacos , Polimorfismo de Fragmento de Restrição , Sensibilidade e Especificidade , Tuberculose Resistente a Múltiplos Medicamentos/microbiologia , Tuberculose Pulmonar/microbiologia
18.
Mol Biol Rep ; 39(2): 919-28, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21562764

RESUMO

Neuropeptide Y (NPY) is a potent orexigenic agent. The molecular mechanisms underlying the regulation of bovine NPY gene expression by its promoter region is currently unknown. The objectives of this research were to: (i) identify the SNPs in the promoter region of the bovine NPY gene, (ii) investigate the effects of these SNPs by measuring promoter transcriptional activities of different bovine NPY promoter haplotypes and; (iii) identify the minimal promoter region (MPR) required for basal activity of the NPY gene in vitro. Seventeen SNPs were identified in the promoter region. Of these, 14 affected putative transcription factors binding motifs including a TATA binding protein factor at -20, GC-Box factors SP1 at -170 and GATA binding motifs at -120 and -347. The SNPs were assigned to five major haplotypes (BtNPY_H1-5), of which BtNPY_H5 had maximum transcriptional activity. The region extending to -134 nt was identified as the MPR. This MPR was confirmed by the identification of a putative TATA box (-29 nt) and two SP1/GC binding sites (-94 and -118 nt), within this region. However, promoter expression was significantly enhanced when the construct contained the -614 to -1019 nt region. In conclusion, a number of SNPs characterised in the bovine NPY promoter especially those affecting the transcription factor binding sites, enhancer and repressor regions have the potential to affect NPY gene expression. Natural variation exists in the promoter region of the bovine NPY gene, which should be further explored for selection of energetic efficiency in cattle.


Assuntos
Bovinos/genética , Regulação da Expressão Gênica/genética , Variação Genética , Neuropeptídeo Y/genética , Polimorfismo de Nucleotídeo Único/genética , Regiões Promotoras Genéticas/genética , Animais , Sequência de Bases , Primers do DNA/genética , Haplótipos/genética , Modelos Lineares , Desequilíbrio de Ligação , Dados de Sequência Molecular , Neuropeptídeo Y/metabolismo , Reação em Cadeia da Polimerase , Domínios e Motivos de Interação entre Proteínas/genética , Análise de Sequência de DNA
19.
Mol Biol Rep ; 39(4): 4411-21, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21947839

RESUMO

The neuropeptide Y 5 receptor (NPY5R) plays an important role in the regulation of appetite and feeding behaviour in mammals by modulating the effect of the neurotransmitter neuropeptide Y. As single nucleotide polymorphism (SNP) variation in the bovine NPY5R gene is likely to influence the expression and/or function of this gene, the objectives of this study were to identify SNPs in the bovine NPY5R gene and to predict their functional role in the expression and physico-chemical characteristics of the protein product. Nineteen novel SNPs were identified in a 2.1 kb genomic region of the NPY5R gene in a total of 419 beef cattle from 13 Bos taurus breeds and eight Bos indicus animals. Four of these SNPs were non-synonymous (Met â†’ Ile, Leu â†’ Phe, Pro â†’ Leu, Arg â†’ Stop codon), while 10 were synonymous. Of particular interest was one non-synonymous SNP (c.1090C>T) that introduced a stop codon in the third intracellular loop of the NPY5R molecule. This stop codon is predicted to create a truncated NPY5R molecule with different physico-chemical properties compared to the native NPY5R protein. A further four SNPs were located in the 5' untranslated region (UTR) and one in the 3'UTR. Two of the 5'UTR SNPs affected putative transcription factor binding sites (GATA binding factor and snRNA-activating protein complex). In conclusion, regulatory and functional SNPs were identified in the bovine NPY5R gene. These include SNPs which potentially modify transcription factor binding sites as well as SNPs that cause amino acid changes and premature termination of the NPY5R protein. Such polymorphisms are likely to play vital physiological roles in the neuropeptide Y mediated appetite, feed intake and energy homeostasis in cattle.


Assuntos
Bovinos/genética , Modelos Moleculares , Polimorfismo de Nucleotídeo Único/genética , Receptores de Neuropeptídeo Y/química , Receptores de Neuropeptídeo Y/genética , Regiões 3' não Traduzidas/genética , Região 5'-Flanqueadora/genética , Regiões 5' não Traduzidas/genética , Animais , Sequência de Bases , Cruzamento , Membrana Celular/metabolismo , Biologia Computacional , Sequência Conservada/genética , Evolução Molecular , Frequência do Gene/genética , Genética Populacional , Haplótipos/genética , Humanos , Dados de Sequência Molecular , Proteínas Mutantes/metabolismo , Nucleotídeos/genética , Fases de Leitura Aberta/genética , Peptídeos/metabolismo
20.
Br J Nutr ; 106(8): 1142-53, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21736830

RESUMO

Recent studies have suggested that chito-oligosaccharides can have anti-adipogenic properties. The objectives of the present study were to evaluate the anti-adipogenic potential of four different chito-oligosaccharides (molecular weight (MW) < 1000, 1000-3000, 3000-5000 and 5000-10,000 Da) and to identify molecular mechanisms underlying the chito-oligosaccharide-mediated inhibition of adipogenesis. Mouse 3T3-L1 cells were allowed to differentiate in the presence of chito-oligosaccharide. At day 8 post-induction of differentiation, lipid accumulation, free glycerol release and the quantitative expression of adipogenic marker genes were evaluated. Chito-oligosaccharides had concentration- and MW-dependent inhibitory effects on lipid accumulation (P < 0·001 and < 0·05, respectively), as well as a concentration-dependent effect (P < 0·001) on free glycerol release and the expression of adipogenic marker genes. The 5000-10,000 Da chito-oligosaccharide was selected for subsequent molecular studies. A panel of forty-four lipid metabolic pathway-specific genes was analysed by quantitative real-time PCR. Chito-oligosaccharide-mediated inhibition of adipogenesis was associated with the up-regulation of the IL-6 gene at all concentrations of chito-oligosaccharide examined and the PG-endoperoxide synthase 2 (PTGS2) gene at higher concentrations of chito-oligosaccharide. The effect of chito-oligosaccharide on gene expression was validated by measuring IL-6 protein concentrations in the media. Finally, an IL-6 promoter assay was developed to characterise the effect of chito-oligosaccharide on the transcriptional activity of the IL-6 promoter, which was increased in a concentration-dependent manner (P < 0·001). We conclude that IL-6 is a candidate signalling molecule in the chito-oligosaccharide-mediated inhibition of adipogenesis in 3T3-L1 cells.


Assuntos
Adipogenia/efeitos dos fármacos , Adipogenia/fisiologia , Interleucina-6/fisiologia , Oligossacarídeos/farmacologia , Células 3T3-L1 , Adipócitos/citologia , Adipócitos/efeitos dos fármacos , Adipócitos/fisiologia , Adipogenia/genética , Animais , Sequência de Bases , Primers do DNA/genética , Expressão Gênica/efeitos dos fármacos , Interleucina-6/genética , Metabolismo dos Lipídeos/efeitos dos fármacos , Metabolismo dos Lipídeos/genética , Redes e Vias Metabólicas/efeitos dos fármacos , Redes e Vias Metabólicas/genética , Camundongos , Peso Molecular , Oligossacarídeos/química , Regiões Promotoras Genéticas , Reação em Cadeia da Polimerase em Tempo Real , Transcriptoma/efeitos dos fármacos
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