RESUMO
Staphylococcus aureus is a species of Gram-positive staphylococcus. It can cause sinusitis, respiratory infections, skin infections, and food poisoning. Recently, it was discovered that S. aureus infects epithelial cells, but the interaction between S. aureus and the host is not well known. In this study, we confirmed S. aureus to be internalized by HaCaT cells using the ESAT-6-like protein EsxB and amplified within the host over time by escaping host immunity. S. aureus increases the expression of decay-accelerating factor (CD55) on the surfaces of host cells, which inhibits the activation of the complement system. This mechanism makes it possible for S. aureus to survive in host cells. S. aureus, sufficiently amplified within the host, is released through the initiation of cell death. On the other hand, the infected host cells increase their surface expression of UL16 binding protein 1 to inform immune cells that they are infected and try to be eliminated. These host defense systems seem to involve the alteration of tight junctions and the induction of ligand expression to activate immune cells. Taken together, our study elucidates a novel aspect of the mechanisms of infection and immune system evasion for S. aureus.
Assuntos
Antígenos CD55/metabolismo , Ativação do Complemento , Staphylococcus aureus/fisiologia , Animais , Proteínas de Bactérias/metabolismo , Morte Celular , Endocitose , Células HaCaT , Interações Hospedeiro-Patógeno , Humanos , Masculino , Camundongos Endogâmicos BALB C , Proteínas de Junções Íntimas/metabolismo , Junções Íntimas/metabolismo , Regulação para Cima/genéticaRESUMO
We previously showed that Lactiplantibacillus plantarum K8 and its cell wall components have immunoregulatory effects. In this study, we demonstrate that pre-treatment of L. plantarum K8 lysates reduced LPS-induced TNF-α production in THP-1 cells by down-regulating the early signals of mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB). The down-regulation of signals may be caused by the induction of negative regulators involved in toll-like receptor (TLR)-mediated signaling. However, co-treatment with high concentrations of L. plantarum K8 lysates and lipopolysaccharide (LPS) activated the late signaling of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and NF-κB pathways and resulted in the induction of absent in melanoma 2 (AIM2) inflammasome-mediated interleukin (IL)-1ß secretion. Intraperitoneal injection of L. plantarum K8 lysates in LPS-induced endotoxin shock mice alleviated mortality and reduced serum tumor-necrosis factor (TNF)-α, IL-1ß, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels. In addition, the mRNA levels of TNF-α, IL-1ß, and IL-6 decreased in livers from mice injected with L. plantarum K8 followed by LPS. Hematoxylin and eosin (H&E) staining of the liver showed that the cell size was enlarged by LPS injection and slightly reduced by L. plantarum K8 lysate pre-injection followed by LPS injection. Macrophage infiltration of the liver also decreased in response to the combination injection compared with mice injected with only LPS. Taken together, our results show that although L. plantarum K8 lysates differentially regulated the production of LPS-induced inflammatory cytokines in THP-1 cells, the lysates inhibited overall inflammation in mice. Thus, this study suggests that L. plantarum K8 lysates could be developed as a substance that modulates immune homeostasis by regulating inflammation.
Assuntos
Inflamação/genética , Lactobacillaceae/química , Fígado/efeitos dos fármacos , Choque Séptico/genética , Animais , Proteínas de Ligação a DNA/genética , Endotoxinas/toxicidade , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Inflamação/induzido quimicamente , Inflamação/patologia , Interleucina-1beta/genética , Interleucina-6/genética , Proteínas Quinases JNK Ativadas por Mitógeno/genética , Lipopolissacarídeos/farmacologia , Lipopolissacarídeos/toxicidade , Fígado/metabolismo , Fígado/patologia , Camundongos , NF-kappa B/genética , Choque Séptico/induzido quimicamente , Choque Séptico/patologia , Fator de Necrose Tumoral alfa/genéticaRESUMO
Toward the standardization of common evening primrose (Oenothera biennis) sprout extract (OBS-E), we aimed to obtain indicator compounds and use a validated method. HPLC-UVD allowed simultaneous quantification of the indicator compounds quercetin-3-O-glucuronide and ellagic acid. The method was validated in terms of specificity, linearity, precision, accuracy, and limit of detection/limit of quantification (LOD/LOQ). High specificity and linearity was demonstrated, with correlation coefficients of 1.0000 for quercetin-3-O-glucuronide and 0.9998 for ellagic acid. The LOD/LOQ values were 0.486/1.472 µg/mL for quercetin-3-O-glucuronide and 1.003/3.039 µg/mL for ellagic acid. Intra-day and inter-day variability tests produced relative standard deviation for each compound of <2%, a generally accepted precision criterion. High recovery rate were also obtained, indicating accuracy validation. The OBS-E prepared using various concentrations of ethanol were then analyzed. The 50% ethanol extract had highest content of quercetin-3-O-glucuronide, whereas the 70% ethanol extract possessed the lowest. However, the ellagic acid content was highest in the 70% ethanol extract and lowest in the 90% ethanol extract. Thus, quercetin-3-O-glucuronide and ellagic acid can be used industrially as indicator compounds for O. biennis sprout products, and our validated method can be used to establish indicator compounds for other natural products.
Assuntos
Ácido Elágico/química , Oenothera biennis/química , Quercetina/análogos & derivados , Cromatografia Líquida de Alta Pressão , Estrutura Molecular , Extratos Vegetais/química , Quercetina/química , Análise EspectralRESUMO
Lipoteichoic acid (LTA), a cell wall component of gram-positive bacteria, up-regulates inflammatory cytokine production through the toll-like receptor 2 (TLR2) signaling pathway, and also contributes to anti-inflammatory responses against immune cells stimulated by lipopolysaccharides. In the current study, we examined the effects of LTAs isolated from Staphylococcus aureus (aLTA) and Lactobacillus plantarum (pLTA) on the aggravation and alleviation of atopic dermatitis (AD). aLTA strongly induced CCL2 production in THP-1 cells. CCL2 was regulated by the TLR2 pathway including the activation of IRAK2, NF-κB and JNK. CCL2 induced Th2 polarization of CD4+T cells through induction of interleukin (IL)-2, -4, and -5 and inhibition of interferon-gamma (IFN-γ). CCL2 levels and immunoglobulin E (IgE) production were increased in aLTA-injected mice. On the other hand, pLTA moderately affected CCL2 production and it inhibited aLTA-mediated CCL2 production. The serum levels of CCL2 and IgE were inhibited by pLTA pre-injection followed by aLTA reinjection, which resulted in the alleviation of irritant contact dermatitis (ICD) symptoms. Our results suggest that S. aureus infection causes an increase in CCL2 production, and may exacerbate atopic dermatitis (AD)-like symptoms through the excessive IgE production. Alternatively, pLTA alleviated AD-like symptoms by inhibiting aLTA-induced CCL2 and IgE production.
Assuntos
Dermatite Atópica , Lactobacillus plantarum , Animais , Lipopolissacarídeos , Camundongos , Staphylococcus aureus , Ácidos TeicoicosRESUMO
Human skin is the first defense barrier against the external environment, especially microbial pathogens and physical stimulation. Many studies on skin health with Lactic acid bacteria (LAB) have been published for many years, including prevention of skin disease and improvement of skin conditions. LAB, a major group of gram-positive bacteria, are known to be beneficial to human health by acting as probiotics. Recent studies have shown that LAB and their extracts have beneficial effects on maintenance and improvement of skin health. Oral administration of Lactobacillus delbrueckii inhibits the development of atopic disease. In addition, LAB and LAB extracts are known to have beneficial effects on intestinal diseases, with Lactobacillus plantarum having been shown to attenuate IL-10 deficient colitis. In addition to intestinal health, L. plantarum also has beneficial effects on skin. pLTA, which is lipoteichoic acid isolated from L. plantarum, has anti-photoaging effects on human skin cells by regulating the expression matrix meralloprotionase-1 (MMP-1) expression. While several studies have proposed a relationship between diseases of the skin and small intestines, there are currently no published reviews of the effects of LAB for skin health through regulation of intestinal conditions and the immune system. In this review, we discuss recent findings on the effects of LAB on skin health and its potential applications in beauty foods.
Assuntos
Lactobacillus/fisiologia , Probióticos/farmacologia , Dermatopatias/prevenção & controle , Extratos Celulares , Alimento Funcional/microbiologia , HumanosRESUMO
Methanol extract of Zizania latifolia was partitioned with EtOAc, n-BuOH, and H2O. From the EtOAc layers, a new flavonolignan along with a known flavone and three known flavonolignans, tricin (1), salcolin A (2), salcolin B (3), and salcolin C (4), were isolated through repeated silica gel and ODS column chromatography. The chemical structure of the new flavonolignan was determined to be tricin-4'-O-[erythro-ß-guaiacyl-(7â³-O-methyl)-glyceryl] ether and was named salcolin D (5) based on physicochemical and spectroscopic data, including FT-NMR and ESI-MS. All compounds were isolated for the first time from this plant. Compounds 2-5, tricin derivatives, all exhibited higher anti-inflammatory and anti-allergy activities than tricin. In particular, salcolin D (5) was shown to have the strongest inhibitory activity against LPS-induced NO production in RAW 264.7 cells as well as ß-hexosaminidase release in IgE-sensitized RBL-2H3 cells. These results suggest that the presence of tricin derivatives conveys allergy and inflammation treatment ability to Z. latifolia.
Assuntos
Antialérgicos/farmacologia , Anti-Inflamatórios não Esteroides/farmacologia , Flavonoides/química , Poaceae/química , Animais , Antialérgicos/química , Anti-Inflamatórios não Esteroides/química , Linhagem Celular/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos/métodos , Flavonas/química , Flavonas/farmacologia , Flavonoides/farmacologia , Flavonolignanos/química , Flavonolignanos/isolamento & purificação , Flavonolignanos/farmacologia , Imunoglobulina E/farmacologia , Lignanas/química , Lignanas/farmacologia , Lipopolissacarídeos/farmacologia , Camundongos , Estrutura Molecular , Óxido Nítrico/metabolismo , Componentes Aéreos da Planta/química , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Ratos , beta-N-Acetil-Hexosaminidases/antagonistas & inibidores , beta-N-Acetil-Hexosaminidases/metabolismoRESUMO
Two new flavonolignan glycosides, tricin-4'-O-(threo-ß-guaiacylglyceryl) ether 7''-O-ß-D-glucopyranose (4) and tricin-4'-O-(erythro-ß-guaiacylglyceryl) ether 7''-O-ß-D-glucopyranose (5) were isolated from the roots of Zizania latifolia, together with tricin-7-O-ß-D-glucopyranose (1), tricin-4'-O-(threo-ß-guaiacylglyceryl) ether 7-O-ß-D-glucopyranose (2), and tricin-4'-O-(erythro-ß-guaiacylglyceryl) ether 7-O-ß-D-glucopyranose (3). Their structures were identified on the basis of spectroscopic techniques, including HR-ESI/MS, 1D-NMR (1H, 13C, DEPT), 2D-NMR (gCOSY, gHSQC, gHMBC), and IR spectroscopy.
Assuntos
Flavonolignanos/isolamento & purificação , Glicosídeos/isolamento & purificação , Extratos Vegetais/química , Poaceae/química , Flavonolignanos/química , Glicosídeos/química , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Componentes Aéreos da Planta/química , Raízes de Plantas/químicaRESUMO
Skin hydration is one of the primary aims of beauty and anti-aging treatments. Barley (Hordeum vulgare) and soybean (Glycine max) are major food crops, but can also be used as ingredients for the maintenance of skin health. We developed a natural product-based skin treatment using a barley and soybean formula (BS) incorporating yeast fermentation, and evaluated its skin hydration effects as a dietary supplement in a clinical study. Participants ingested a placebo- (n = 33) or BS- (3 g/day) containing drink (n = 32) for 8 weeks. A significant increase in hydration in the BS group as compared to the placebo group was observed on the faces of subjects after 4 and 8 weeks, and on the forearm after 4 weeks. Decreases in stratum corneum (SC) thickness were also observed on the face and forearm. BS enhanced hyaluronan (HA) and skin barrier function in vitro and reduced Hyal2 expression in human dermal fibroblasts (HDF). BS also recovered ultraviolet (UV) B-induced downregulation of HA in HaCaT cells. These results suggest that BS has promising potential for development as a health functional food to enhance skin health.
RESUMO
Developmentally regulated GTP-binding protein 2 (DRG2) represents a novel subclass of GTP-binding proteins. We here report that transgenic overexpression of DRG2 in mice ameliorates experimental autoimmune encephalomyelitis (EAE), a murine model of multiple sclerosis (MS). The protective effect of DRG2 in EAE was mediated by the inhibition of the development of T(H)17 cells. DRG2 enhanced the activity of PPARγ, which led to an inhibition of the nuclear factor kappa B (NF-κB) activity and IL-6 production in antigen presenting cells and an inhibition of the development of T(H)17 cells. Our results demonstrate that DRG2 is an essential modulator of EAE.
Assuntos
Encefalomielite Autoimune Experimental/genética , Encefalomielite Autoimune Experimental/imunologia , Proteínas de Ligação ao GTP/genética , Células Th17/imunologia , Animais , Células Apresentadoras de Antígenos/imunologia , Células Apresentadoras de Antígenos/metabolismo , Diferenciação Celular , Proteínas Correpressoras/metabolismo , Citocinas/metabolismo , Encefalomielite Autoimune Experimental/metabolismo , Proteínas de Ligação ao GTP/metabolismo , Expressão Gênica , Genótipo , Mediadores da Inflamação/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Masculino , Camundongos , Camundongos Transgênicos , NF-kappa B/metabolismo , PPAR gama/metabolismo , Regiões Promotoras Genéticas , Ligação Proteica , Subpopulações de Linfócitos T/citologia , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Células Th17/citologia , Células Th17/metabolismoRESUMO
Three recombinant polypeptides, VP1-His, VP1-3N-His, and 3D2-His, were produced by Escherichia coli expression system. Recombinant VP1-His, VP1-3N-His, and 3D2-His were expressed as bands with molecular weights of 32, 38, and 30 kDa, respectively. These were purified by affinity chromatography using Ni-NTA Fast-flow resin and/or ion-exchange chromatography using DEAE-Sepharose Fast-flow resin. Intraperitoneal immunizations of recombinant polypeptides successfully elicited the productions of VP1-His, VP1-3N-His, and 3D2-His specific IgG antibodies (IgG subclass distribution of IgG1>IgG2a>IgG2b>IgG3) in sera and induced the secretions of cytokines IFN-γ and IL-6 in spleen cells. Sera from recombinant VP1-His-, VP1-3N-His-, and 3D2-His-immunized mice neutralized the propagation of HAV. The highest neutralizing activity was shown in sera from recombinant VP1-3N-His-immunized mice. These results suggest that recombinant VP1-3N-His can be a useful source for developing hepatitis A virus (HAV) subunit vaccine candidates.
Assuntos
Vírus da Hepatite A/imunologia , Hepatite A/prevenção & controle , Proteínas Recombinantes/imunologia , Proteínas Estruturais Virais/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/imunologia , Proteínas do Capsídeo/isolamento & purificação , Células Cultivadas , Clonagem Molecular , Escherichia coli/genética , Feminino , Hepatite A/sangue , Hepatite A/imunologia , Imunização , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Interferon gama/imunologia , Interleucina-6/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Baço/citologia , Baço/imunologia , Proteínas Estruturais Virais/genética , Proteínas Estruturais Virais/isolamento & purificação , Vacinas Virais/genética , Vacinas Virais/isolamento & purificaçãoRESUMO
OBJECTIVE: Interleukin-6 (IL-6), which is increased in patients who are suffering from septic shock, is an important mediator of the inflammatory response. Here, we examined the priming effect of lipoteichoic acid (LTA) and lipopolysaccharide (LPS) on IL-6 production in a monocyte-like cell line. METHODS: THP-1 cells were primed by treatingwith a low or high dose of LTA isolated from Staphylococcus aureus (aLTA) and then re-treated with LPS. IL-6 production, receptor expression, and the variation of signaling molecules were examined by ELISA, reverse transcriptase polymerase chain reaction, and western blotting, respectively. RESULTS: LPS-mediated IL-6 production was dramatically increased in THP-1 cells pretreated with a low dose aLTA, while it was significantly decreased when a high dose of aLTA was given along with LPS. LPS-induced IL-6 production in low dose aLTA priming cells mediated by NF-κB and MAPKs pathways, and Akt functioned as a negative regulator of IL-6 production. Together, the results of this study suggest that different doses of bacterial cell surface components can mediate a diverse range of responses with respect to inflammatory cytokine production.
Assuntos
Interleucina-6/metabolismo , Lipopolissacarídeos/administração & dosagem , Ácidos Teicoicos/administração & dosagem , Linhagem Celular Tumoral , Humanos , Lactobacillus plantarum , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Staphylococcus aureusRESUMO
Hypoxic responses have been implicated in critical pathologies, including inflammation, immunity, and tumorigenesis. Recently, efforts to identify effective natural remedies and health supplements are increasing. Previous studies have reported that the cell lysates and the cell wall-bound lipoteichoic acids of Lactiplantibacillus plantarum K8 (K8) exert anti-inflammatory and immunomodulative effects. However, the effect of K8 on cellular hypoxic responses remains unknown. In this study, we found that K8 lysates had a potent suppressive effect on gene expression under hypoxia. K8 lysates markedly downregulated hypoxia-induced HIF1α accumulation in the human bone marrow and lung cancer cell lines, SH-SY5Y and H460. Consequently, the transcription of known HIF1α target genes, such as p21, GLUT1, and ALDOC, was notably suppressed in the K8 lysate supplement and purified lipoteichoic acids of K8, upon hypoxic induction. Intriguingly, K8 lysates decreased the expression of PHD2 and VHL proteins, which are responsible for HIF1α destabilization under normoxic conditions, suggesting that K8 may regulate HIF1α stability in a non-canonical pathway. Overall, our results suggest that K8 lysates desensitize the cells to hypoxic stresses and suppress HIF1α-mediated hypoxic gene activation.
Assuntos
Neuroblastoma , Humanos , Hipóxia Celular/genética , Linhagem Celular , Hipóxia/metabolismo , Expressão Gênica , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismoRESUMO
Multiple studies have confirmed that Lactiplantibacillus plantarum has beneficial effects in respiratory diseases, including respiratory tract infections, asthma, and chronic obstructive pulmonary disease. However, the role of L. plantarum lysates in respiratory diseases is unclear. Staphylococcus aureus infects the lungs of mice, recruits immune cells, and induces structural changes in alveoli. Lung diseases can be further aggravated by inflammatory cytokines such as CCL2 and interleukin (IL)-6. In in vivo studies, L. plantarum K8 nanoparticles (K8NPs) restored lung function and prevented lung damage caused by S. aureus infection. They inhibited the S. aureus infection and the infiltration of immune cells and prevented the increase in goblet cell numbers in the lungs of S. aureus-infected mice. K8NPs suppressed the expression of CCL2 and IL-6, which were increased by the combination treatment of tumor necrosis factor alpha and interferon gamma (TI), in a dose-dependent manner. In in vitro studies, the anti-inflammatory effect of K8NPs in TI-treated A549 cells and TI-injected mice occurred through the reduction in activated mitogen-activated protein kinases and nuclear factor kappa-B. These findings suggest that the efficacy of K8NPs in controlling respiratory inflammation and infection can be used to develop functional materials that can prevent or alleviate respiratory diseases.
Assuntos
Lactobacillus plantarum , Pneumonia , Infecções Respiratórias , Infecções Estafilocócicas , Animais , Camundongos , Fator de Necrose Tumoral alfa/metabolismo , Interferon gama/metabolismo , Staphylococcus aureus , Citocinas/metabolismo , Pneumonia/prevenção & controle , Interleucina-6/metabolismo , Infecções Respiratórias/prevenção & controle , Lactobacillus plantarum/metabolismoRESUMO
Probiotics exert anti-obesity effects in high-fat diet (HFD) obese mice, but there are few studies on anti-obesity using heat-killed probiotics. Here, we investigated the effect of heat-killed Lactiplantibacillus plantarum K8 (K8HK) on the anti-differentiation of 3T3-L1 preadipocytes and on anti-obesity in HFD mice. K8HK decreased triglyceride (TG) accumulation in 3T3-L1 cells. Specifically, 1 × 109 CFU/mL K8HK showed the greatest anti-obesity effect, while the same concentration of live L. plantarum K8 (K8 Live) showed cytotoxicity. K8HK increased suppressor of cytokine signaling (SOCS)-1, which might affect the JAK2-STAT3 signaling pathway activated during differentiation. As a result, the levels of transcription factors of adipogenesis such as Peroxisome proliferator-activated receptor γ (PPARγ) and CCAAT/enhancer binding protein α (C/EBPα) decreased in K8HK-treated cells. We also observed a decrease in the lipogenic enzymes and fatty acid binding protein 4 (FABP4). In the mouse study, oral ingestion of K8 Live and K8HK showed weight reduction and decrease in blood TG content at 12 weeks of feeding. In addition, TG synthesis was suppressed in liver and adipose tissues, and genes related to fat metabolism were suppressed. This study suggests that K8HK could be a good material to prevent obesity by inhibiting adipogenesis genes related to fat metabolism.
RESUMO
A phytosterol derivative, 3-O-ß-D-glucopyanosylspinasterol (spinasterol-Glc) isolated from leaves of Stewartia koreana was reported to inhibit LPS-induced cytokine production in macrophage cells. Thymus and activation regulated chemokine (TARC/CCL17) is produced in response to pro-inflammatory cytokines in keratinocytes, which is implicated in the development of inflammatory skin diseases. In present study, we investigated the effect of spinasterol-Glc on production of TARC/CCL17 induced by TNF-α and IFN-γ in human HaCaT keratinocytes. Spinasterol-Glc inhibited the mRNA and protein expression of TARC/CCL17 induced by TNF-α/IFN-γ in a dose-dependent manner. Inhibitors of c-Raf-1, p38 MAPK, and JAK2, suppressed the TNF-α/IFN-γ-induced production of TARC/CCL17, and phosphorylation of these signaling molecules were attenuated by spinasterol-Glc. The compound also inhibited phosphorylation of IKKα/ß and IκB-α, and reduced translocation of NF-κB to the nucleus. We demonstrated that spinasterol-Glc suppressed the NF-κB-driven and the GAS-driven expression of luciferase reporter gene induced by TNF-α and IFN-γ. In addition, spinasterol-Glc inhibited the DNA binding of NF-κB and STAT1 to its cognate binding site. These results suggest that spinasterol-Glc has effective inhibitory effects on production of TARC/CCL17 in keratinocytes via inhibition of NF-κB as well as STAT activation, and could be utilized for development of a potential therapeutic agent against skin inflammatory diseases.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Quimiocina CCL17/antagonistas & inibidores , Fármacos Dermatológicos/farmacologia , Queratinócitos/efeitos dos fármacos , NF-kappa B/antagonistas & inibidores , Fator de Transcrição STAT1/antagonistas & inibidores , Estigmasterol/análogos & derivados , Linhagem Celular , Quimiocina CCL17/biossíntese , Dermatite Atópica/tratamento farmacológico , Dermatite Atópica/metabolismo , Humanos , Interferon gama/farmacologia , Janus Quinase 2/metabolismo , Queratinócitos/metabolismo , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-raf/metabolismo , Transdução de Sinais/efeitos dos fármacos , Estigmasterol/farmacologia , Fator de Necrose Tumoral alfa/farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Lipoteichoic acid isolated from Staphylococcus aureus (aLTA) is known to regulate the production of pro-inflammatory cytokines through TLR2-mediated signaling pathways. In our previous study, we found that aLTA significantly increased manganese superoxide dismutase (MnSOD) in the THP-1 human monocyte-like cell line, but the role of MnSOD in the regulation of cytokine production was not elucidated. In the current study, we found that MnSOD was involved in aLTA-mediated cytokine production. The signaling pathways associated with aLTA-mediated MnSOD induction in THP-1 cells included TLR2-MyD88-IRAK2, JNK (c-Jun N-terminal kinases)1/2 and nuclear factor- κB (NF-κB). We also found MnSOD was involved in the regulation of IL-1ß and TNF-α, which were induced by early signaling pathways, including JNK1/2, p38, and NF-κB p65. In addition, MnSOD was also involved in the production of IL-6 and CCL2 in aLTA-stimulated THP-1 cells through activation of late signaling pathways such as JAK2-STAT3. Taken together, our data suggest that aLTA-mediated MnSOD production involved in the regulation of cytokine production and it may be the cause of one of the excessive inflammatory reactions caused by S. aureus.
Assuntos
Citocinas , Staphylococcus aureus , Citocinas/metabolismo , Humanos , Lipopolissacarídeos , Staphylococcus aureus/metabolismo , Superóxido Dismutase , Células THP-1 , Ácidos TeicoicosRESUMO
Lipoteichoic acid is a major cell wall virulence factor of gram-positive bacteria. LTAs from various bacteria have differential immunostimulatory potentials due to heterogeneity in their structures. Although recent studies have demonstrated that LTA isolated from Lactobacillus plantarum (pLTA) has anti-inflammatory properties and is less inflammatory than LTAs from pathogenic bacteria, little is known about the structure of pLTA. In this study, high-field NMR spectra of the pLTA were compared with those of LTA from pathogenic bacterium, Staphylococcus aureus (aLTA). The 2D NMR results demonstrated that pLTA possesses α-linked hexose sugar substituents on the poly-glycerophosphate backbone instead of N-acetylglucosamine substituents, and unsaturated fatty acids in its glycolipids. The sugar substituents were revealed as an approximately 29:1 molar ratio of the glucose to galactose by HPAEC-PAD analysis. MALDI-TOF/TOF MS analyses identified the presence of unsaturated fatty acids in the glycolipid moieties of pLTA. In addition, the glycolipid structure was found to be composed of trihexosyl-diacyl- and/or trihexosyl-triacyl-glycerol ceramide units by means of unique fragment ions of the glycolipids. These results enabled us to elucidate the pLTA structure, which is distinctively different from canonical LTA structure, and suggest that the unique immunological property of pLTA might be caused by the pLTA structure.
Assuntos
Lactobacillus plantarum/química , Lipopolissacarídeos/química , Ácidos Teicoicos/química , Lipopolissacarídeos/isolamento & purificação , Espectroscopia de Ressonância Magnética , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Staphylococcus aureus/química , Ácidos Teicoicos/isolamento & purificaçãoRESUMO
Methanol extract of Koreana stewartia leaves (SKE) stimulated collagen production in ultraviolet-B (UVB)-irradiated human fibroblast cells. An active compound was isolated from SKE by successive partitioning and chromatography, and the chemical structure was determined to be 3-O-ß-D-glucopyranosylspinasterol (spinasterol-Glc) by spectroscopic characterization. Spinasterol-Glc increased collagen production in the supernatant of UVB-irradiated dermal fibroblast cell cultures in a dose-dependent manner. The effects of spinasteol-Glc on expression of procollagen and matrix metalloproteinase-1 (MMP-1) were further evaluated. We found that the compound stimulated collagen production in UVB-treated fibroblasts than in vehicle-treated control cells by about 3-fold. In addition, we also demonstrate that the compound increased the mRNA and protein levels of procollagen in UVB-treated fibroblast cells, while it inhibited expression of MMP-1. These results indicate that spinasterol-Glc protects fibroblast cells from the adverse effects of UV radiation via stimulation of procollagen synthesis as well as inhibition of MMP-1 expression. Spinasterol-Glc may be useful in the future development of therapeutic and cosmetic applications.
Assuntos
Metaloproteinase 1 da Matriz/metabolismo , Pró-Colágeno/biossíntese , Pele/efeitos da radiação , Estigmasterol/análogos & derivados , Theaceae/química , Sequência de Bases , Western Blotting , Células Cultivadas , Primers do DNA , Fibroblastos/enzimologia , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Humanos , Pele/enzimologia , Pele/metabolismo , Espectrometria de Massas de Bombardeamento Rápido de Átomos , Estigmasterol/isolamento & purificação , Estigmasterol/farmacologiaRESUMO
Most of the studies on common evening primrose (Oenothera biennis L.) are focused on its oils (isolated from seed, root, and stem tissues). We aimed to investigate the effect of the oral administration of OBS-E on the improvement of skin function in skin-damaged hairless mice exposed to excessive ultraviolet B (UVB) radiation owing to the preliminary in vitro findings regarding the antioxidant, anti-wrinkle, and skin moisturizing activities of OBS-E. OBS-E administration for 14 weeks did not significantly affect the body weight or clinical signs. Significant reductions were observed in wrinkle parameters (area, number, length, and depth, and metalloproteinase levels) in OBS-E-administered mice compared with those in UVB-irradiated control mice. OBS-E significantly increased skin elasticity and hyaluronic acid content, but it significantly decreased transepidermal water loss. Histomorphometrical analysis revealed that OBS-E significantly reduced the epidermal thickness, area of the collagen-occupied region, and number of microfolds and inflammatory and mast cells. These results demonstrate that OBS-E can effectively enhance skin functions in terms of ameliorating wrinkle formation, promoting skin-moisturization, enhancing skin barrier function, and inhibiting inflammatory reactions. The obtained results provide good starting point for the continuation in the process of developing new inner beauty products based on OBS-E.
RESUMO
Staphylococcus aureus inhibits complement activity by secreting a variety of toxins. However, the underlying mechanism of complement component regulation by lipoteichoic acid (LTA), a cell wall component of S. aureus, has not been elucidated. In this study, we observed that aLTA (LTA of S. aureus) increased C3 expression in THP-1 cells. The mechanism of aLTA-mediated C3 induction includes an aLTA-toll-like receptor (TLR) 2 interaction, interleukin 1 receptor associated kinase (IRAK) 2 recruitment, and nuclear factor kappa B (NF-kB) activation. In HepG2 cells, C3 protein production begins to increase from 3 h and increases steadily until 48 h. On the other hand, CD55 levels increased up to 6 h after aLTA treatment and started to decrease after 24 h and levels were decreased at 48 h by more than 50% compared to untreated cells. The expression of CD55 in HepG2 cells was shown to be regulated by IRAK-M induced by aLTA. Serum C3 levels increased in mice injected with aLTA, which resulted in an increase in the amount and activity of the membrane attack complex (MAC). We also observed that CD55 mRNA was increased in the liver 24 h after aLTA injection, but was decreased 48 h after injection. These results suggest that aLTA increases complement levels via induction of C3 and inhibition of CD55, which may cause associated MAC-mediated liver damage.