RESUMEN
LPS binding protein (LBP) is an important innate sensor of microbial cell wall structures. Frequent functionally relevant mutations exist and have been linked to influence susceptibility to and course of bacterial infections. We examined functional properties of a single nucleotide polymorphism resulting in an exchange of phenylalanine to leucine at position 436 of LBP (rs2232618) and compared the frequent variant of the molecule with the rare one in ligand binding experiments. We then stimulated RAW cells with bacterial ligands in the presence of serum obtained from individuals with different LBP genotypes. We, furthermore, determined the potential effects of structural changes in the molecule by in silico modeling. Finally, we analyzed 363 surgical patients for this genetic variant and examined incidence and course of sepsis following surgery. We found that binding of LBP to bacterial ligands was reduced, and stimulation of RAW cells resulted in an increased release of TNF when adding serum from individuals carrying the F436L variant as compared with normal LBP. In silico analysis revealed structural changes of LBP, potentially explaining some of the effects observed for the LBP variant. Finally, patients carrying the F436L variant were found to be similarly susceptible for sepsis. However, we observed a more favorable course of severe infections in this cohort. Our findings reveal new insights into LPS recognition and the subsequent activation of the innate immune system brought about by LBP. The identification of a genetic variant of LBP influencing the course of sepsis may help to stratify individuals at risk and thus reduce clinical complications of patients.
Asunto(s)
Proteínas de Fase Aguda/genética , Proteínas de Fase Aguda/fisiología , Proteínas Portadoras/genética , Proteínas Portadoras/fisiología , Variación Genética/genética , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/fisiología , Sepsis/genética , Sepsis/inmunología , Animales , Línea Celular , Simulación por Computador , Genotipo , Humanos , Ratones , Polimorfismo de Nucleótido SimpleRESUMEN
OBJECTIVES: To explore the role of LPS binding protein (LBP) in metabolism and optimize sepsis treatment. DESIGN: A sepsis model was established by injecting LPS into LBP-/- rats and WT rats and observing changes in the liver over time (0, 1, 6, and 24âh). SETTING: Detecting liver inflammation and injury. Optimizing the treatment of sepsis. SUBJECTS: WT rats and LBP-/- rats. INTERVENTIONS: We established a sepsis model by injecting LPS intravenously. MEASUREMENTS AND MAIN RESULTS: First, we induced sepsis in WT and LBP-/- rats with LPS. The rats were sacrificed, and serum and liver samples were collected at 1, 6, and 24âh after LPS injection. We found that the deletion of LBP reduced LPS-induced liver inflammation and injury at 1 and 6âh. Ballooning degeneration was clearly present in LBP-/- rat livers at 24âh after LPS injection. We found that mitochondrial damage and reactive oxygen species (ROS) levels were higher in LBP-/- rat livers than in WT rat livers at 24âh after LPS injection. According to the transcriptomic results, the peroxisome proliferator-activated receptor (PPAR) pathway may be the reason for lesions in LBP-/- rats. To further investigate the function of PPARα in sepsis, we inhibited mTOR with rapamycin and examined mitochondrial injury and ROS levels. The levels of mitochondrial damage and ROS were reduced after LBP-/- rats were pretreated with rapamycin in the context of LPS-induced sepsis. Inhibiting CYP4a2, one of the PPARα-target gene products, reduced the level of LPS-induced ROS in LBP-/- rats. CONCLUSION: LBP protects hepatic mitochondria against LPS-induced damage via the LBP-PPARα-CYP4a2 signaling pathway.
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Proteínas de Fase Aguda/fisiología , Proteínas Portadoras/fisiología , Sistema Enzimático del Citocromo P-450/fisiología , Glicoproteínas de Membrana/fisiología , Mitocondrias Hepáticas/metabolismo , Receptores Activados del Proliferador del Peroxisoma/fisiología , Sepsis/metabolismo , Transducción de Señal/fisiología , Animales , Modelos Animales de Enfermedad , RatasRESUMEN
OBJECTIVE: Pulmonary fibrosis is a fatal interstitial lung disease that is characterized by excessive accumulation of extracellular matrix (ECM) and remodeling of lung. The precise mechanisms underlying pulmonary fibrosis still remain unclear. In the current study, we aimed to investigate the alteration and function of serine (or cysteine) peptidase inhibitor, clade A, member 3 N (Serpina3n) in pulmonary fibrotic models and explore the potential mechanisms. METHODS: We induced pulmonary fibrosis in mice by silica and bleomycin respectively and determined Serpina3n in lung tissues, and then verified the expression of Serpina3n and its correlation with pulmonary fibrosis at seven time points in a bleomycin longstanding model. Moreover, adeno-associated virus type 9 (AAV9)-mediated Serpina3n knockdown was used to treat pulmonary fibrosis in the bleomycin model, whose possible mechanisms would be preliminarily explored by detecting chymotrypsin C as an example. RESULTS: Serpina3n was up-regulated significantly in lungs of both models at mRNA and protein levels relative to control. Notably, the expression of Serpina3n peaked during the 3rd week and then decreased until nearly normal levels during the 10th week, which was closely related to fibrotic procession in bleomycin-treated mice. AAV-mediated Serpina3n knockdown in the lung tissues alleviated bleomycin-induced fibrotic symptoms at various levels and disinhibit chymotrypsin C. CONCLUSIONS: Our study revealed that Serpina3n is a critical regulator in pulmonary fibrosis and suggested Serpina3n inhibition as a potential therapeutic strategy in chronic pulmonary injuries.
Asunto(s)
Proteínas de Fase Aguda/fisiología , Fibrosis Pulmonar/metabolismo , Serpinas/fisiología , Proteínas de Fase Aguda/genética , Proteínas de Fase Aguda/metabolismo , Animales , Bleomicina , Quimotripsina/metabolismo , Técnicas de Silenciamiento del Gen , Ratones , Fibrosis Pulmonar/inducido químicamente , Fibrosis Pulmonar/enzimología , Fibrosis Pulmonar/patología , Serpinas/genética , Serpinas/metabolismo , Regulación hacia ArribaRESUMEN
OBJECTIVE: Cancer cachexia and muscle loss are associated with increased morbidity and mortality. In preclinical animal models, blocking activin receptor (ACVR) ligands has improved survival and prevented muscle wasting in cancer cachexia without an effect on tumour growth. However, the underlying mechanisms are poorly understood. This study aimed to identify cancer cachexia and soluble ACVR (sACVR) administration-evoked changes in muscle proteome. METHODS: Healthy and C26 tumour-bearing (TB) mice were treated with recombinant sACVR. The sACVR or PBS control were administered either prior to the tumour formation or by continued administration before and after tumour formation. Muscles were analysed by quantitative proteomics with further examination of mitochondria and nicotinamide adenine dinucleotide (NAD+) metabolism. To complement the first prophylactic experiment, sACVR (or PBS) was injected as a treatment after tumour cell inoculation. RESULTS: Muscle proteomics in TB cachectic mice revealed downregulated signatures for mitochondrial oxidative phosphorylation (OXPHOS) and increased acute phase response (APR). These were accompanied by muscle NAD+ deficiency, alterations in NAD+ biosynthesis including downregulation of nicotinamide riboside kinase 2 (Nrk2), and decreased muscle protein synthesis. The disturbances in NAD+ metabolism and protein synthesis were rescued by treatment with sACVR. Across the whole proteome and APR, in particular, Serpina3n represented the most upregulated protein and the strongest predictor of cachexia. However, the increase in Serpina3n expression was associated with increased inflammation rather than decreased muscle mass and/or protein synthesis. CONCLUSIONS: We present evidence implicating disturbed muscle mitochondrial OXPHOS proteome and NAD+ homeostasis in experimental cancer cachexia. Treatment of TB mice with a blocker of activin receptor ligands restores depleted muscle NAD+ and Nrk2, as well as decreased muscle protein synthesis. These results indicate putative new treatment therapies for cachexia and that although acute phase protein Serpina3n may serve as a predictor of cachexia, it more likely reflects a condition of elevated inflammation.
Asunto(s)
Proteínas de Fase Aguda/metabolismo , Músculo Esquelético/metabolismo , NAD/metabolismo , Serpinas/metabolismo , Receptores de Activinas/antagonistas & inhibidores , Receptores de Activinas/efectos de los fármacos , Receptores de Activinas/metabolismo , Activinas/metabolismo , Activinas/farmacología , Proteínas de Fase Aguda/fisiología , Animales , Caquexia/metabolismo , Caquexia/fisiopatología , Línea Celular Tumoral , Modelos Animales de Enfermedad , Masculino , Ratones , Mitocondrias/metabolismo , Músculo Esquelético/fisiología , Atrofia Muscular/metabolismo , Miostatina/metabolismo , Fosforilación Oxidativa , Serpinas/fisiologíaRESUMEN
Hypercoagulability has been recognized as a common complication of COVID-19. Exact mechanisms for this extreme coagulation activation have not yet been elucidated. However, one of the consistent laboratory finding is the increase in fibrinogen, in some cases, marked elevation. High circulating levels of fibrinogen have been linked to thrombosis for years and for this reason, hyperfibrinogenemia is considered one of the mechanisms for COVID-19 coagulopathy. In this forum article, instead of the prothrombotic role, a protective function for fibrinogen is discussed. Fibrinogen, like the other well-known acute phase reactants, is increased in COVID-19 possibly to protect the host.
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Betacoronavirus , Infecciones por Coronavirus/sangre , Fibrinógeno/fisiología , Inflamación/sangre , Neumonía Viral/sangre , Trombofilia/sangre , Trombosis/prevención & control , Enfermedad Aguda , Proteínas de Fase Aguda/fisiología , Biomarcadores , Plaquetas/fisiología , COVID-19 , Infecciones por Coronavirus/complicaciones , Productos de Degradación de Fibrina-Fibrinógeno/análisis , Productos de Degradación de Fibrina-Fibrinógeno/fisiología , Fibrinógeno/análisis , Humanos , Modelos Cardiovasculares , Pandemias , Neumonía Viral/complicaciones , Riesgo , SARS-CoV-2 , Trombofilia/etiología , Factor de von Willebrand/análisis , Factor de von Willebrand/fisiologíaRESUMEN
During the aging process, bone marrow mesenchymal stem cells (BMSCs) exhibit declined osteogenesis accompanied by excess adipogenesis, which will lead to osteoporosis. Here, we report that the H3 lysine 36 trimethylation (H3K36me3), catalyzed by histone methyltransferase SET-domain-containing 2 (SETD2), regulates lineage commitment of BMSCs. Deletion of Setd2 in mouse bone marrow mesenchymal stem cells (mBMSCs), through conditional Cre expression driven by Prx1 promoter, resulted in bone loss and marrow adiposity. Loss of Setd2 in BMSCs in vitro facilitated differentiation propensity to adipocytes rather than to osteoblasts. Through conjoint analysis of RNA sequencing (RNA-seq) and chromatin immunoprecipitation sequencing (ChIP-seq) data, we identified a SETD2 functional target gene, Lbp, on which H3K36me3 was enriched, and its expression was affected by Setd2 deficiency. Furthermore, overexpression of lipopolysaccharide-binding protein (LBP) could partially rescue the lack of osteogenesis and enhanced adipogenesis resulting from the absence of Setd2 in BMSCs. Further mechanistic studies demonstrated that the trimethylation level of H3K36 could regulate Lbp transcriptional initiation and elongation. These findings suggest that H3K36me3 mediated by SETD2 could regulate the cell fate of mesenchymal stem cells (MSCs) in vitro and in vivo, indicating that the regulation of H3K36me3 level by targeting SETD2 and/or the administration of downstream LBP may represent a potential therapeutic way for new treatment in metabolic bone diseases, such as osteoporosis.
Asunto(s)
Células de la Médula Ósea/fisiología , N-Metiltransferasa de Histona-Lisina/fisiología , Histonas/genética , Proteínas de Fase Aguda/genética , Proteínas de Fase Aguda/fisiología , Adipocitos/citología , Animales , Proteínas Portadoras/genética , Proteínas Portadoras/fisiología , Diferenciación Celular/genética , Metilación de ADN/genética , N-Metiltransferasa de Histona-Lisina/metabolismo , Lisina/metabolismo , Masculino , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/fisiología , Células Madre Mesenquimatosas/fisiología , Ratones , Ratones Endogámicos C57BL , Regiones Promotoras Genéticas/genéticaRESUMEN
While torpor is a beneficial energy-saving strategy, it may incur costs if an animal is unable to respond appropriately to external stimuli, which is particularly true when it is necessary to escape from threats such as fire. We aimed to determine whether torpid bats, which are potentially threatened because they must fly to escape, can sense smoke and whether respiration rate (RR), heart rate (HR) and reaction time of torpid bats prior to and following smoke introduction is temperature-dependent. To test this we quantified RR and HR of captive Australian tree-roosting bats, Nyctophilus gouldi (n=5, ~10g), in steady-state torpor in response to short-term exposure to smoke from Eucalyptus spp. leaves between ambient temperatures (Ta) of 11 and 23°C. Bats at lower Ta took significantly longer (28-fold) to respond to smoke, indicated by a cessation of episodic breathing and a rapid increase in RR. Bats at lower Ta returned to torpor more swiftly following smoke exposure than bats at higher Ta. Interestingly, bats at Ta<15°C never returned to thermoconforming steady-state torpor prior to the end of the experimental day, whereas all bats at Ta≥15°C did, as indicated by apnoeic HR. This shows that although bats at lower Ta took longer to respond, they appear to maintain vigilance and prevent deep torpor after the first smoke exposure, likely to enable fast escape. Our study reveals that bats can respond to smoke stimuli while in deep torpor. These results are particularly vital within the framework of fire management conducted at Ta<15°C, as most management burns are undertaken during winter when bats will likely respond more slowly to fire cues such as smoke, delaying the time to escape from the fire.
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Quirópteros/fisiología , Percepción Olfatoria/fisiología , Humo , Letargo/fisiología , Proteínas de Fase Aguda/fisiología , Animales , Reacción de Fuga/fisiología , Eucalyptus , Frecuencia Cardíaca/fisiología , Masculino , Movimiento/fisiología , Hojas de la Planta , Respiración , TemperaturaRESUMEN
Toll-like receptor 4 (TLR4) together with MD2, one of the key pattern recognition receptors for a pathogen-associated molecular pattern, activates innate immunity by recognizing lipopolysaccharide (LPS) of Gram-negative bacteria. Although LBP and CD14 catalyze LPS transfer to the TLR4/MD2 complex, the detail mechanisms underlying this dynamic LPS transfer remain elusive. Using negative-stain electron microscopy, we visualized the dynamic intermediate complexes during LPS transfer-LBP/LPS micelles and ternary CD14/LBP/LPS micelle complexes. We also reconstituted the entire cascade of LPS transfer to TLR4/MD2 in a total internal reflection fluorescence (TIRF) microscope for a single molecule fluorescence analysis. These analyses reveal longitudinal LBP binding to the surface of LPS micelles and multi-round binding/unbinding of CD14 to single LBP/LPS micelles via key charged residues on LBP and CD14. Finally, we reveal that a single LPS molecule bound to CD14 is transferred to TLR4/MD2 in a TLR4-dependent manner. These discoveries, which clarify the molecular mechanism of dynamic LPS transfer to TLR4/MD2 via LBP and CD14, provide novel insights into the initiation of innate immune responses. [BMB Reports 2017; 50(2): 55-57].
Asunto(s)
Proteínas de Fase Aguda/fisiología , Proteínas Portadoras/fisiología , Receptores de Lipopolisacáridos/fisiología , Lipopolisacáridos/metabolismo , Antígeno 96 de los Linfocitos/metabolismo , Glicoproteínas de Membrana/fisiología , Receptor Toll-Like 4/metabolismo , Proteínas de Fase Aguda/química , Proteínas de Fase Aguda/metabolismo , Animales , Transporte Biológico , Proteínas Portadoras/química , Proteínas Portadoras/metabolismo , Humanos , Inmunidad Innata , Receptores de Lipopolisacáridos/química , Receptores de Lipopolisacáridos/metabolismo , Lipopolisacáridos/química , Lipopolisacáridos/inmunología , Antígeno 96 de los Linfocitos/química , Glicoproteínas de Membrana/química , Glicoproteínas de Membrana/metabolismo , Modelos Moleculares , Complejos Multiproteicos/química , Complejos Multiproteicos/metabolismo , Unión Proteica , Transducción de Señal/inmunología , Receptor Toll-Like 4/químicaRESUMEN
BACKGROUND AND AIM: It has been suggested in several studies that an increased translocation of bacterial lipopolysaccharide (LPS) and, subsequently, an activation of toll-like receptor (TLR)-dependent signaling pathways in the liver may contribute to the development of non-alcoholic fatty liver disease. METHODS: Eight-week-old lipopolysaccharide-binding protein (LBP)-/- and wild-type (WT) mice were pair fed either a liquid diet rich in fat, fructose, and cholesterol (Western-style diet [WSD]) or a control liquid diet for 8 weeks. Parameters of liver injury, markers of TLR-4-dependent signaling pathway, and glucose/lipid metabolism were determined. RESULTS: Despite similar total caloric intake, weight gain, fasting blood glucose levels, and liver-to-bodyweight ratio, indices of liver damage determined by liver histology and transaminases were markedly lower in WSD-fed LBP-/- mice than in WSD-fed WT animals. In line with these findings, number of neutrophils, F4/80 positive cells, and plasminogen activator inhibitor 1 were only found to be significantly increased in livers of WSD-fed WT mice. While mRNA expressions of TLR-4 and myeloid differentiation primary response 88 were similar between WSD-fed groups, concentrations of inducible nitric oxide synthase protein and 4-hydroxynonenal protein adducts were significantly higher in livers of WSD-fed WT mice than in WSD-fed LBP-/- animals. Markers of lipid metabolism, for example, sterol regulatory element-binding protein 1c and fatty acid synthase per se, were significantly lower in livers of LBP-/- mice; however, mRNA expressions did not differ between controls and WSD-fed mice within the respective mouse strain. CONCLUSION: Taken together, our results suggest that LBP is a critical factor in the development of non-alcoholic fatty liver disease in mice.
Asunto(s)
Proteínas de Fase Aguda/deficiencia , Proteínas de Fase Aguda/fisiología , Proteínas Portadoras/fisiología , Lipopolisacáridos/metabolismo , Hígado/metabolismo , Glicoproteínas de Membrana/deficiencia , Glicoproteínas de Membrana/fisiología , Enfermedad del Hígado Graso no Alcohólico/etiología , Animales , Modelos Animales de Enfermedad , Glucosa/metabolismo , Metabolismo de los Lípidos , Ratones Endogámicos BALB C , Ratones Noqueados , Enfermedad del Hígado Graso no Alcohólico/genética , Transducción de Señal/fisiología , Receptores Toll-Like/metabolismoRESUMEN
OBJECTIVE: Cross-sectional studies suggest that lipopolysaccharide-binding protein (LBP) may be associated with obesity and metabolic disorders. However, prospective studies examining LBP are lacking. This prospective study investigated the association between LBP and metabolic abnormalities in 580 African ancestry men (mean age, 59.1 ± 10.5 years). RESEARCH DESIGN AND METHODS: We measured fasting serum LBP at baseline. Changes in adiposity and glucose homeostasis as well as case subjects with new type 2 diabetes and impaired fasting glucose (IFG) were assessed at a follow-up visit ~6 years later. Baseline LBP values were tested across quartiles for linear trend with metabolic measures. Multivariable logistic regression was used to determine the odds of new cases of IFG or diabetes per 1-SD greater baseline LBP. RESULTS: LBP was significantly associated with baseline BMI, waist circumference, whole-body and trunk fat, skeletal muscle density, fasting serum insulin, and HOMA-insulin resistance (IR) (all P < 0.01). Greater baseline LBP was significantly associated with longitudinal increases in the percentage of trunk fat (P = 0.025) and HOMA-IR (P = 0.034), but only borderline so with a decrease in skeletal muscle density (P = 0.057). In men with normal glucose, baseline LBP was associated with increased odds of having IFG at follow-up after adjustment for age, baseline trunk fat, and lifestyle factors (odds ratio per 1-SD LBP: 1.51; 95% CI 1.02-2.21). This association was attenuated after additional adjustment for change in trunk fat (P = 0.067). CONCLUSIONS: LBP may be a marker of prediabetes. Some of this association appears to be mediated through increased central and ectopic skeletal muscle adiposity.
Asunto(s)
Adiposidad/fisiología , Proteínas Portadoras/sangre , Glicoproteínas de Membrana/sangre , Estado Prediabético/sangre , Proteínas de Fase Aguda/fisiología , Adulto , Anciano , Envejecimiento/sangre , Envejecimiento/fisiología , Biomarcadores/sangre , Población Negra , Glucemia/análisis , Glucemia/metabolismo , Proteínas Portadoras/fisiología , Estudios Transversales , Humanos , Masculino , Glicoproteínas de Membrana/fisiología , Persona de Mediana Edad , Obesidad/sangre , Obesidad/fisiopatología , Estado Prediabético/fisiopatología , Estudios ProspectivosRESUMEN
The tubular lipid-binding (TULIP) superfamily has emerged in recent years as a major mediator of lipid sensing and transport in eukaryotes. It currently encompasses three protein families, SMP-like, BPI-like, and Takeout-like, which share a common fold. This fold consists of a long helix wrapped in a highly curved anti-parallel ß-sheet, enclosing a central, lipophilic cavity. The SMP-like proteins, which include subunits of the ERMES complex and the extended synaptotagmins (E-Syts), appear to be mainly located at membrane contacts sites (MCSs) between organelles, mediating inter-organelle lipid exchange. The BPI-like proteins, which include the bactericidal/permeability-increasing protein (BPI), the LPS (lipopolysaccharide)-binding protein (LBP), the cholesteryl ester transfer protein (CETP), and the phospholipid transfer protein (PLTP), are either involved in innate immunity against bacteria through their ability to sense lipopolysaccharides, as is the case for BPI and LBP, or in lipid exchange between lipoprotein particles, as is the case for CETP and PLTP. The Takeout-like proteins, which are comprised of insect juvenile hormone-binding proteins and arthropod allergens, transport, where known, lipid hormones to target tissues during insect development. In all cases, the activity of these proteins is underpinned by their ability to bind large, hydrophobic ligands in their central cavity and segregate them away from the aqueous environment. Furthermore, where they are involved in lipid exchange, recent structural studies have highlighted their ability to establish lipophilic, tubular channels, either between organelles in the case of SMP domains or between lipoprotein particles in the case of CETP. Here, we review the current knowledge on the structure, versatile functions, and evolution of the TULIP superfamily. We propose a deep evolutionary split in this superfamily, predating the Last Eukaryotic Common Ancestor, between the SMP-like proteins, which act on lipids endogenous to the cell, and the BPI-like proteins (including the Takeout-like proteins of arthropods), which act on exogenous lipids. This article is part of a Special Issue entitled: The cellular lipid landscape edited by Tim P. Levine and Anant K. Menon.
Asunto(s)
Células Eucariotas/metabolismo , Proteínas de Unión a Ácidos Grasos/fisiología , Metabolismo de los Lípidos , Proteínas de Fase Aguda/química , Proteínas de Fase Aguda/fisiología , Animales , Péptidos Catiónicos Antimicrobianos/química , Péptidos Catiónicos Antimicrobianos/fisiología , Transporte Biológico/genética , Proteínas Sanguíneas/química , Proteínas Sanguíneas/fisiología , Proteínas Portadoras/química , Proteínas Portadoras/fisiología , Proteínas de Transferencia de Ésteres de Colesterol/química , Proteínas de Transferencia de Ésteres de Colesterol/fisiología , Proteínas de Unión a Ácidos Grasos/química , Humanos , Metabolismo de los Lípidos/genética , Glicoproteínas de Membrana/química , Glicoproteínas de Membrana/fisiología , Modelos Moleculares , Familia de Multigenes/fisiología , Proteínas de Transferencia de Fosfolípidos/química , Proteínas de Transferencia de Fosfolípidos/fisiología , FilogeniaRESUMEN
BACKGROUND AND AIM: Because neutrophil gelatinase-associated lipocalin (NGAL) is known to provide significant bacteriostatic effects during infectious conditions, we tested the hypothesis that this protein is up-regulated and secreted into the intraluminal cavity of the gut under critically ill conditions and is thus responsible for the regulation of bacterial overgrowth. METHODS: With our institutional approval, male C57BL/6J mouse (6-7 weeks) were enrolled and applied for lipopolysaccharide or peritonitis model compared with naïve control. We assessed NGAL protein concentrations in intestinal lumen and up-regulation of NGAL expression in intestinal tissues in in vivo as well as ex vivo settings. Simultaneously, we examined the effects of NGAL protein administration on the growth of Escherichia coli (E. coli) in in vivo and in vitro experimental settings. The localization of NGAL in intestinal tissues and lumen was also assessed by immunohistological approach using NGAL antibody. RESULTS: Both lipopolysaccharide and peritonitis insults evoked the marked up-regulation of NGAL mRNA and protein levels in gut tissues such as crypt cells. In addition, the administration of NGAL protein significantly inhibited the outgrowth of enteric E. coli under both in vitro and in vivo conditions, accompanied by histological evidence. CONCLUSION: Neutrophil gelatinase-associated lipocalin protein accompanied by apparent bacteriostatic action accumulated in the intestinal wall and streamed into the mucosal layer during critically ill state, thereby possibly shaping microbiota homeostasis in the gut.
Asunto(s)
Proteínas de Fase Aguda/farmacología , Proteínas de Fase Aguda/fisiología , Intestinos/microbiología , Lipocalinas/farmacología , Lipocalinas/fisiología , Microbiota/efectos de los fármacos , Proteínas Oncogénicas/farmacología , Proteínas Oncogénicas/fisiología , Proteínas de Fase Aguda/genética , Proteínas de Fase Aguda/metabolismo , Animales , Enfermedad Crítica , Modelos Animales de Enfermedad , Escherichia coli/crecimiento & desarrollo , Expresión Génica , Homeostasis/efectos de los fármacos , Mucosa Intestinal/metabolismo , Lipocalina 2 , Lipocalinas/genética , Lipocalinas/metabolismo , Lipopolisacáridos , Masculino , Ratones Endogámicos C57BL , Microbiota/fisiología , Proteínas Oncogénicas/genética , Proteínas Oncogénicas/metabolismo , Peritonitis/microbiología , Regulación hacia ArribaRESUMEN
Chronic itch is an intractable symptom of inflammatory skin diseases, such as atopic and contact dermatitis. Recent studies have revealed neuronal pathways selective for itch, but the mechanisms by which itch turns into a pathological chronic state are poorly understood. Using mouse models of atopic and contact dermatitis, we demonstrate a long-term reactive state of astrocytes in the dorsal horn of the spinal segments that corresponds to lesioned, itchy skin. We found that reactive astrogliosis depended on the activation of signal transducer and activator of transcription 3 (STAT3). Conditional disruption of astrocytic STAT3 suppressed chronic itch, and pharmacological inhibition of spinal STAT3 ameliorated the fully developed chronic itch. Mice with atopic dermatitis exhibited an increase in scratching elicited by intrathecal administration of the itch-inducer gastrin-releasing peptide (GRP), and this enhancement was normalized by suppressing STAT3-mediated reactive astrogliosis. Moreover, we identified lipocalin-2 (LCN2) as an astrocytic STAT3-dependent upregulated factor that was crucial for chronic itch, and we demonstrated that intrathecal administration of LCN2 to normal mice increased spinal GRP-evoked scratching. Our findings indicate that STAT3-dependent reactive astrocytes act as critical amplifiers of itching through a mechanism involving the enhancement of spinal itch signals by LCN2, thereby providing a previously unrecognized target for treating chronic itch.
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Prurito/etiología , Factor de Transcripción STAT3/fisiología , Asta Dorsal de la Médula Espinal/patología , Proteínas de Fase Aguda/fisiología , Animales , Astrocitos/fisiología , Enfermedad Crónica , Péptido Liberador de Gastrina/fisiología , Lipocalina 2 , Lipocalinas/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Proteínas Oncogénicas/fisiologíaRESUMEN
PURPOSE: The current study was done to determine the role of lipocalin-2 (LCN2) in the pathogenesis of demyelinating optic neuritis using an experimental autoimmune optic neuritis (EAON) model. METHODS: The EAON was induced by subcutaneous immunization with an emulsified mixture of myelin oligodendrocyte glycoprotein (MOG35-55) peptide in mice. The LCN2 expression was examined in the optic nerve after MOG peptide injection. Degree of demyelination, inflammatory infiltration, glial activation, and expression profile of inflammatory mediators in the optic nerve were compared between LCN2 knockout (KO) animals and wild-type littermates by histological analysis and real-time PCR following EAON induction. Plasma levels of LCN2 in patients with optic neuritis were measured by ELISA. RESULTS: The expression of LCN2 was notably increased in the optic nerve after EAON induction. Expression of LCN2 was colocalized with reactive astrocytes. A significant reduction of demyelination, inflammatory infiltration, and gliosis was demonstrated in the optic nerve of LCN2 KO mice. The LCN2 KO mice also showed markedly reduced gene expression associated with the M1-polarized glia phenotype and toll-like receptor signaling in the optic nerve. The LCN2 levels in plasma were significantly higher in optic neuritis patients (71.6 ± 10.6 ng/mL) compared to healthy controls (37.4 ± 9.1 ng/mL, P = 0.0284). CONCLUSIONS: In this study, we demonstrated a significant induction of LCN2 expression in astrocytes of the optic nerve following EAON induction. Our results imply that astrocyte-derived LCN2 may have a pivotal role in the development of demyelinating optic neuritis, and LCN2 can be a therapeutic target to alleviate immune and inflammatory damage in the optic nerve.
Asunto(s)
Proteínas de Fase Aguda/fisiología , Astrocitos , Enfermedades Desmielinizantes/etiología , Lipocalinas/fisiología , Neuritis Óptica/etiología , Proteínas Proto-Oncogénicas/fisiología , Proteínas de Fase Aguda/biosíntesis , Adulto , Animales , Astrocitos/metabolismo , Enfermedades Desmielinizantes/sangre , Femenino , Humanos , Lipocalina 2 , Lipocalinas/biosíntesis , Lipocalinas/sangre , Masculino , Ratones , Ratones Endogámicos C57BL , Neuritis Óptica/sangre , Proteínas Proto-Oncogénicas/biosíntesis , Proteínas Proto-Oncogénicas/sangreRESUMEN
Activation of the mineralocorticoid receptor has been shown to be deleterious in cardiovascular diseases (CVDs). We have recently shown that lipocalin 2 (Lcn2), or neutrophil gelatinase-associated lipocalin (NGAL), is a primary target of aldosterone/mineralocorticoid receptor in the cardiovascular system. Lcn2 is a circulating protein, which binds matrix metalloproteinase 9 and modulates its stability. We hypothesized that Lcn2 could be a mediator of aldosterone/mineralocorticoid receptor profibrotic effects in the cardiovascular system. Correlations between aldosterone and profibrotic markers, such as procollagen type I N-terminal peptide, were investigated in healthy subjects and subjects with abdominal obesity. The implication of Lcn2 in the mineralocorticoid pathway was studied using Lcn2 knockout mice subjected to a nephrectomy/aldosterone/salt (NAS) challenge for 4 weeks. In human subjects, NGAL/matrix metalloproteinase 9 was positively correlated with plasma aldosterone and fibrosis biomarkers. In mice, loss of Lcn2 prevented the NAS-induced increase of plasma procollagen type I N-terminal peptide, as well as the increase of collagen fibers deposition and collagen I expression in the coronary vessels and the aorta. The lack of Lcn2 also blunted the NAS-induced increase in systolic blood pressure. Ex vivo, treatment of human fibroblasts with recombinant Lcn2 induced the expression of collagen I and the profibrotic galectin-3 and cardiotrophin-1 molecules. Our results showed that Lcn2 plays a key role in aldosterone/mineralocorticoid receptor-mediated vascular fibrosis. The clinical data indicate that this may translate in human patients. Lcn2 is, therefore, a new biotarget in cardiovascular fibrosis induced by mineralocorticoid activation.
Asunto(s)
Proteínas de Fase Aguda/fisiología , Aldosterona/toxicidad , Lipocalinas/fisiología , Obesidad Abdominal/fisiopatología , Proteínas Oncogénicas/fisiología , Proteínas Proto-Oncogénicas/fisiología , Proteínas de Fase Aguda/deficiencia , Proteínas de Fase Aguda/genética , Proteínas de Fase Aguda/farmacología , Aldosterona/sangre , Animales , Aorta/efectos de los fármacos , Aorta/patología , Cardiomiopatía Hipertrófica/inducido químicamente , Cardiomiopatía Hipertrófica/fisiopatología , Células Cultivadas , Citocinas/biosíntesis , Citocinas/genética , Femenino , Fibroblastos , Fibrosis , Galectina 3/biosíntesis , Galectina 3/sangre , Galectina 3/genética , Humanos , Hipertensión/fisiopatología , Hipertrofia , Riñón/patología , Lipocalina 2 , Lipocalinas/sangre , Lipocalinas/genética , Lipocalinas/farmacología , Masculino , Ratones , Miocardio/citología , Miocardio/metabolismo , Miocitos del Músculo Liso/efectos de los fármacos , Miocitos del Músculo Liso/fisiología , Nefrectomía/efectos adversos , Obesidad Abdominal/sangre , Proteínas Oncogénicas/deficiencia , Proteínas Oncogénicas/genética , Fragmentos de Péptidos/sangre , Procolágeno/sangre , Proteínas Proto-Oncogénicas/sangre , Proteínas Proto-Oncogénicas/farmacología , Ratas , Proteínas Recombinantes/farmacologíaAsunto(s)
Acné Vulgar/tratamiento farmacológico , Coristoma/tratamiento farmacológico , Fármacos Dermatológicos/uso terapéutico , Isotretinoína/uso terapéutico , Enfermedades de los Labios/tratamiento farmacológico , Glándulas Sebáceas , Acné Vulgar/complicaciones , Proteínas de Fase Aguda/biosíntesis , Proteínas de Fase Aguda/fisiología , Adulto , Coristoma/complicaciones , Coristoma/patología , Fármacos Dermatológicos/farmacología , Humanos , Isotretinoína/farmacología , Enfermedades de los Labios/complicaciones , Enfermedades de los Labios/patología , Lipocalina 2 , Lipocalinas/biosíntesis , Lipocalinas/fisiología , Masculino , Proteínas Proto-Oncogénicas/biosíntesis , Proteínas Proto-Oncogénicas/fisiología , Glándulas Sebáceas/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacosRESUMEN
BACKGROUND: Lipocalin-2 is an adipocytokine implicated in apoptosis, innate immunity, angiogenesis, and the development of chronic kidney disease. OBJECTIVES: To investigate the role of lipocalin-2 in systemic sclerosis (SSc). MATERIALS AND METHODS: Serum lipocalin-2 levels were determined by enzyme-linked immunosorbent assay in 50 patients with SSc and 19 healthy subjects. Lipocalin-2 expression was evaluated in the skin of patients with SSc and bleomycin (BLM)-treated mice and in Fli1-deficient endothelial cells by reverse transcriptase-real time polymerase chain reaction, immunoblotting and/or immunohistochemistry. RESULTS: Although serum lipocalin-2 levels were comparable between patients with SSc and healthy controls, the prevalence of scleroderma renal crisis was significantly higher in patients with SSc with elevated serum lipocalin-2 levels than in those with normal levels. Furthermore, serum lipocalin-2 levels inversely correlated with estimated glomerular filtration rate in patients with SSc with renal dysfunction. Among patients with SSc with normal renal function, serum lipocalin-2 levels positively correlated with skin score in patients with diffuse cutaneous SSc with disease duration of < 3 years and inversely correlated with estimated right ventricular systolic pressure in total patients with SSc. Importantly, in SSc lesional skin, lipocalin-2 expression was increased in dermal fibroblasts and endothelial cells. In BLM-treated mice, lipocalin-2 was highly expressed in dermal fibroblasts, but not in endothelial cells. On the other hand, the deficiency of transcription factor Fli1, which is implicated in SSc vasculopathy, induced lipocalin-2 expression in cultivated endothelial cells. CONCLUSIONS: Lipocalin-2 may be involved in renal dysfunction and dermal fibrosis of SSc. Dysregulated matrix metalloproteinase-9/lipocalin-2-dependent angiogenesis due to Fli1 deficiency may contribute to the development of pulmonary arterial hypertension associated with SSc.
Asunto(s)
Proteínas de Fase Aguda/fisiología , Lipocalinas/fisiología , Enfermedades Pulmonares/etiología , Proteínas Proto-Oncogénicas/fisiología , Insuficiencia Renal Crónica/etiología , Esclerodermia Sistémica/etiología , Piel/patología , Enfermedades Vasculares/etiología , Proteínas de Fase Aguda/metabolismo , Adulto , Anciano , Animales , Apoptosis/fisiología , Estudios de Casos y Controles , Femenino , Fibrosis/etiología , Fibrosis/patología , Fibrosis/fisiopatología , Tasa de Filtración Glomerular/fisiología , Humanos , Lipocalina 2 , Lipocalinas/metabolismo , Enfermedades Pulmonares/fisiopatología , Masculino , Ratones , Persona de Mediana Edad , Proteínas Proto-Oncogénicas/metabolismo , Esclerodermia Sistémica/patología , Esclerodermia Sistémica/fisiopatología , Enfermedades Cutáneas Vasculares/etiología , Enfermedades Cutáneas Vasculares/fisiopatología , Enfermedades Vasculares/patología , Enfermedades Vasculares/fisiopatologíaRESUMEN
BACKGROUND: Ethylene glycol (EG) exposure caused formation of calcium oxalate crystal that led to renal failure, which is associated with higher prevalence of hypertension. L-Arginine is known to have an antioxidant and nephro-protective potential. OBJECTIVE: To evaluate the effect of L-arginine against EG-induced urolithiasis in uninephrectomized hypertensive rats. MATERIAL AND METHODS: Uninephrectomized male Wistar rats (180-200 g) were used to induce urinary calculi through oral administration of EG (0.75%) in distilled water. Rats were treated with either distilled water (10 mg/kg, p.o.) or telmisartan (10 mg/kg, p.o.) or Cystone (500 mg/kg, p.o.) or L-arginine (250, 500, and 1000 mg/kg, p.o.) for 28 days. Various hemodynamic, biochemical, molecular, and histological parameters were assessed in kidney and heart. RESULTS: Rats treated with L-arginine (500 and 1000 mg/kg) significantly restored altered relative organ weight, urine output, urine density, urinary pH, and water intake. EG-induced alterations in electrocardiographic (QRS interval, HR, and ST height) and hemodynamic (SBP, DBP, MABP, and LVEDP) abnormalities were significantly restored by L-arginine (500 and 1000 mg/kg) treatment. It also significantly restored alteration in serum and urine biochemical parameters induced by EG. The elevated oxido-nitrosative stress was also significantly decreased by L-arginine (500 and 1000 mg/kg) treatment. It also significantly down-regulated EG-induced up-regulated renal KIM-1, NGAL, eNOS, and iNOs mRNA expressions. Histological aberrations induced in the renal and cardiac tissues were also ameliorated by l-arginine treatment. CONCLUSION: L-Arginine exerts its nephro- and cardio-protective potential in EG-induced urolithiasis in uninephrectomized hypertensive rats via modulation of KIM-1, NGAL, eNOS, and iNOs mRNA expression.
Asunto(s)
Arginina/uso terapéutico , Urolitiasis/tratamiento farmacológico , Proteínas de Fase Aguda/fisiología , Animales , Moléculas de Adhesión Celular/fisiología , Glicol de Etileno/administración & dosificación , Lipocalina 2 , Lipocalinas/fisiología , Masculino , Óxido Nítrico Sintasa de Tipo III/fisiología , Proteínas Proto-Oncogénicas/fisiología , Ratas , Ratas Endogámicas SHR , Ratas Wistar , Urolitiasis/inducido químicamenteRESUMEN
NGAL (neutrophil gelatinase-associated lipocalin) is an acute phase protein, participating in antibacterial immunity. NGAL forms a complex with metalloproteinase 9 (MMP-9), thereby increasing its activity and preventing its degradation. NGAL is freely filtered through the glomerular membrane and reabsorbed by endocytosis in the proximal tubule. NGAL detected in urine is produced mainly in the distal nephron. Elevated serum and urine NGAL allows diagnosis of acute kidney injury approximately 24 hours earlier than plasma creatinine concentration. Increased levels of NGAL were detected in patients with acute myocardial infarction, heart failure or stroke and were demonstrated to be strong predictors of adverse prognosis.