ABSTRACT
NLRP3-inflammasome-driven inflammation is involved in the pathogenesis of a variety of diseases. Identification of endogenous inflammasome activators is essential for the development of new anti-inflammatory treatment strategies. Here, we identified that apolipoprotein C3 (ApoC3) activates the NLRP3 inflammasome in human monocytes by inducing an alternative NLRP3 inflammasome via caspase-8 and dimerization of Toll-like receptors 2 and 4. Alternative inflammasome activation in human monocytes is mediated by the Toll-like receptor adapter protein SCIMP. This triggers Lyn/Syk-dependent calcium entry and the production of reactive oxygen species, leading to activation of caspase-8. In humanized mouse models, ApoC3 activated human monocytes in vivo to impede endothelial regeneration and promote kidney injury in an NLRP3- and caspase-8-dependent manner. These data provide new insights into the regulation of the NLRP3 inflammasome and the pathophysiological role of triglyceride-rich lipoproteins containing ApoC3. Targeting ApoC3 might prevent organ damage and provide an anti-inflammatory treatment for vascular and kidney diseases.
Subject(s)
Acute Kidney Injury/immunology , Apolipoprotein C-III/immunology , Caspase 8/metabolism , Kidney Diseases/immunology , Monocytes/immunology , NLR Family, Pyrin Domain-Containing 3 Protein/immunology , Acute Kidney Injury/pathology , Adaptor Proteins, Signal Transducing , Animals , Apolipoprotein C-III/genetics , Cell Line , Disease Models, Animal , HEK293 Cells , Humans , Inflammasomes/immunology , Inflammation/genetics , Inflammation/immunology , Kidney Diseases/pathology , Membrane Proteins , Mice , Mice, Inbred C57BL , Mice, Inbred NOD , Mice, Knockout , Reactive Oxygen Species/metabolism , Toll-Like Receptor 2/metabolism , Toll-Like Receptor 4/metabolismABSTRACT
Nicotinamide phosphoribosyltransferase (NAMPT) is the rate-limiting enzyme in the NAD+ salvage pathway from nicotinamide. By controlling the biosynthesis of NAD+, NAMPT regulates the activity of NAD+-converting enzymes, such as CD38, poly-ADP-ribose polymerases, and sirtuins (SIRTs). SIRT6 is involved in the regulation of a wide number of metabolic processes. In this study, we investigated the ability of SIRT6 to regulate intracellular NAMPT activity and NAD(P)(H) levels. BxPC-3 cells and MCF-7 cells were engineered to overexpress a catalytically active or a catalytically inactive SIRT6 form or were engineered to silence endogenous SIRT6 expression. In SIRT6-overexpressing cells, NAD(H) levels were up-regulated, as a consequence of NAMPT activation. By immunopurification and incubation with recombinant SIRT6, NAMPT was found to be a direct substrate of SIRT6 deacetylation, with a mechanism that up-regulates NAMPT enzymatic activity. Extracellular NAMPT release was enhanced in SIRT6-silenced cells. Also glucose-6-phosphate dehydrogenase activity and NADPH levels were increased in SIRT6-overexpressing cells. Accordingly, increased SIRT6 levels reduced cancer cell susceptibility to H2O2-induced oxidative stress and to doxorubicin. Our data demonstrate that SIRT6 affects intracellular NAMPT activity, boosts NAD(P)(H) levels, and protects against oxidative stress. The use of SIRT6 inhibitors, together with agents inducing oxidative stress, may represent a promising treatment strategy in cancer.-Sociali, G., Grozio, A., Caffa, I., Schuster, S., Becherini, P., Damonte, P., Sturla, L., Fresia, C., Passalacqua, M., Mazzola, F., Raffaelli, N., Garten, A., Kiess, W., Cea, M., Nencioni, A., Bruzzone, S. SIRT6 deacetylase activity regulates NAMPT activity and NAD(P)(H) pools in cancer cells.
Subject(s)
Cytokines/metabolism , NADP/metabolism , Neoplasms/metabolism , Nicotinamide Phosphoribosyltransferase/metabolism , Sirtuins/metabolism , Cell Line , Cell Line, Tumor , Doxorubicin/pharmacology , Glucosephosphate Dehydrogenase/metabolism , HEK293 Cells , Hep G2 Cells , Humans , Hydrogen Peroxide/pharmacology , MCF-7 Cells , Neoplasms/drug therapy , Oxidative Stress/drug effects , Oxidative Stress/physiology , Poly(ADP-ribose) Polymerases/metabolism , Up-Regulation/drug effects , Up-Regulation/physiologyABSTRACT
The NLRP3 inflammasome, a caspase-1 activation platform, plays a key role in the modulation of liver inflammation and fibrosis. Here, we tested the hypothesis that interleukin 17 (IL-17) and tumor necrosis factor (TNF) are key cytokines involved in amplifying and perpetuating the liver damage and fibrosis resulting from NLRP3 activation. To address this hypothesis, gain-of-function Nlrp3A350V knock-in mice were bred onto il17a and Tnf knockout backgrounds allowing for constitutive Nlrp3 activation in myeloid derived cells in mice deficient in IL-17 or TNF. Livers of Nlrp3A350V knock-in mice exhibited severe liver inflammatory changes characterized by infiltration with neutrophils, increased expression of chemokine (C-X-C motif) ligand (CXCL) 1 and CXCL2 chemokines, activated inflammatory macrophages, and elevated levels of IL-17 and TNF. Mutants with ablation of il17a signal showed fewer neutrophils when compared to intact Nlrp3A350V mutants, but still significant inflammatory changes when compared to the nonmutant il17a knockout littermates. The severe inflammatory changes associated with mutant Nlrp3 were almost completely rescued by Tnf knockout in association with a marked decrease in circulating IL-1ß levels. Intact Nlrp3A350V mutants showed changes in liver fibrosis, as evidenced by morphometric quantitation of Sirius Red staining and increased mRNA levels of profibrotic genes, including connective tissue growth factor and tissue inhibitor of matrix metalloproteinase 1. Il17a lacking mutants exhibited amelioration of the aforementioned fibrosis, whereas Tnf-deficient mutants showed no signs of fibrosis when compared to littermate controls. Conclusion: Our study uncovers key roles for TNF and, to a lesser extent, IL-17 as mediators of liver inflammation and fibrosis induced by constitutive NLRP3 inflammasome activation in myeloid-derived cells. These findings may lead to therapeutic strategies aimed at halting the progression of liver injury and fibrogenesis in various liver pathogeneses driven by NLRP3 activation. (Hepatology 2018;67:736-749).
Subject(s)
Hepatitis/etiology , Interleukin-17/physiology , Liver Cirrhosis, Experimental/etiology , NLR Family, Pyrin Domain-Containing 3 Protein/physiology , Tumor Necrosis Factor-alpha/physiology , Animals , Hepatic Stellate Cells/physiology , Macrophages/physiology , Mice , Neutrophil Infiltration , Signal TransductionABSTRACT
Circulating oxidized linoleic acid (LA) metabolites (OXLAMs) are increased in patients with nonalcoholic steatohepatitis (NASH) and their levels correlate with disease severity. However, the mechanisms by which OXLAMs contribute to NASH development are incompletely understood. We tested the hypothesis that LA or OXLAMs provided directly through the diet are involved in the development of hepatic injury. C57BL/6 mice were fed an isocaloric high-fat diet containing low LA, high LA, or OXLAMs for 8 weeks. The livers of OXLAM-fed mice showed lower triglyceride concentrations, but higher FA oxidation and lipid peroxidation in association with increased oxidative stress. OXLAM-induced mitochondrial dysfunction was associated with reduced Complex I protein and hepatic ATP levels, as well as increased mitochondrial biogenesis and cytoplasmic mitochondrial DNA. Oxidative stress increased thioredoxin-interacting protein (TXNIP) in the liver and stimulated the activation of mitochondrial apoptosis signal-regulating kinase 1 (ASK1) leading to apoptosis. We also found increased levels of NOD-like receptor protein 3 (NLRP3) inflammasome components and Caspase-1 activation in the livers of OXLAM-fed mice. In vitro, OXLAMs induced hepatocyte cell death, which was partly dependent on Caspase-1 activation. This study identified key mechanisms by which dietary OXLAMs contribute to NASH development, including mitochondrial dysfunction, hepatocyte cell death, and NLRP3 inflammasome activation.
Subject(s)
Apoptosis/drug effects , Linoleic Acid/metabolism , Linoleic Acid/pharmacology , Mitochondria, Liver/drug effects , Mitochondria, Liver/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Animals , Body Weight/drug effects , Carrier Proteins/metabolism , Diet, High-Fat/adverse effects , Gene Expression Regulation/drug effects , Inflammasomes/metabolism , Lipid Peroxidation/drug effects , Male , Mice , Mice, Inbred C57BL , Oxidative Stress/drug effects , Thioredoxins/metabolismABSTRACT
Alcoholic liver disease is a major human health problem leading to significant morbidity and mortality in the United States and worldwide. Dietary fat plays an important role in alcoholic liver disease pathogenesis. Herein, we tested the hypothesis that a combination of ethanol and a diet rich in linoleic acid (LA) leads to the increased production of oxidized LA metabolites (OXLAMs), specifically 9- and 13-hydroxyoctadecadienoic acids (HODEs), which contribute to a hepatic proinflammatory response exacerbating liver injury. Mice were fed unsaturated (with a high LA content) or saturated fat diets (USF and SF, respectively) with or without ethanol for 10 days, followed by a single binge of ethanol. Compared to SF+ethanol, mice fed USF+ethanol had elevated plasma alanine transaminase levels, enhanced hepatic steatosis, oxidative stress, and inflammation. Plasma and liver levels of 9- and 13-HODEs were increased in response to USF+ethanol feeding. We demonstrated that primarily 9-HODE, but not 13-HODE, induced the expression of several proinflammatory cytokines in vitro in RAW264.7 macrophages. Finally, deficiency of arachidonate 15-lipoxygenase, a major enzyme involved in LA oxidation and OXLAM production, attenuated liver injury and inflammation caused by USF+ethanol feeding but had no effect on hepatic steatosis. This study demonstrates that OXLAM-mediated induction of a proinflammatory response in macrophages is one of the potential mechanisms underlying the progression from alcohol-induced steatosis to alcoholic steatohepatitis.
Subject(s)
Dietary Fats/adverse effects , Inflammation/pathology , Linoleic Acid/adverse effects , Liver/metabolism , Liver/pathology , Animals , Arachidonate 15-Lipoxygenase/metabolism , Binge Drinking , Body Composition , Cytokines/metabolism , Disease Models, Animal , Ethanol , Linoleic Acids/metabolism , Linoleic Acids, Conjugated/metabolism , Macrophages/metabolism , Metabolome , Mice , Mice, Inbred C57BL , Oxidation-Reduction , Oxidative Stress , RAW 264.7 CellsABSTRACT
Circular RNAs (circRNAs) are covalently closed single-stranded RNA molecules derived from exons by alternative mRNA splicing. Circularization of single-stranded RNA molecules was already described in 1976 for viroids in plants. Since then several additional types of circular RNAs in many species have been described such as the circular single-stranded RNA genome of the hepatitis delta virus (HDV) or circular RNAs as products or intermediates of tRNA and rRNA maturation in archaea. CircRNAs are generally formed by covalent binding of the 5' site of an upstream exon with the 3' of the same or a downstream exon. Meanwhile, two different models of circRNA biogenesis have been described, the lariat or exon skipping model and the direct backsplicing model. In the lariat model, canonical splicing occurs before backsplicing, whereas in the direct backsplicing model, the circRNA is generated first. In this chapter, we will review the formation of circular RNAs and highlight the derivation of different types of circular RNAs.
Subject(s)
Alternative Splicing , RNA/genetics , Exons/genetics , Gene Expression Regulation , Humans , Introns/genetics , Models, Genetic , Nucleic Acid Conformation , RNA/metabolism , RNA, Bacterial/genetics , RNA, Bacterial/metabolism , RNA, Circular , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , RNA, Viral/genetics , RNA, Viral/metabolism , Spliceosomes/metabolism , Viroids/geneticsABSTRACT
BACKGROUND: Obese individuals differ in their risk of developing metabolic and cardiovascular complications depending on fat distribution (subcutaneous versus visceral) and adipose tissue (AT) phenotype (hyperplasic versus hypertrophic). However, the exact mechanisms which determine whether an obese individual is metabolically healthy or unhealthy are not clear, and analyses of the underlying pathomechanisms are limited by the lack of suitable in vivo models in which metabolically healthy versus metabolically unhealthy AT accumulation can be specifically induced. In the current study, we aimed to establish a protocol for the use of zebrafish as a model for obesity-related metabolically healthy versus metabolically unhealthy AT accumulation. METHODS: We overfed adult male zebrafish of the AB strain with normal fat diet (NFD) or high fat diet (HFD) for 8 weeks and compared parameters related to obesity, i.e. body weight, body mass index, condition index and body fat percentage, to control zebrafish fed under physiological conditions. In addition, we investigated the presence of early obesity-related metabolic alterations by quantifying blood glucose levels, plasma triglyceride and cholesterol levels, and by assessing ectopic lipid accumulation in the liver of zebrafish. Finally, we determined gene expression levels of marker genes related to lipid metabolism, inflammation and fibrosis in visceral AT and liver. RESULTS: We show that 8-weeks overfeeding with either NFD or HFD leads to a significant increase in body weight and AT mass compared to controls. In contrast to NFD-overfed zebrafish, HFD-overfed zebrafish additionally present metabolic alterations, e.g. hyperglycemia and ectopic lipid accumulation in the liver, and a metabolically unhealthy AT phenotype with adipocyte hypertrophy especially in the visceral AT depot, which is accompanied by changes in the expression of marker genes for lipid metabolism, inflammation and fibrosis. CONCLUSIONS: In summary, we have established a method for the specific induction of metabolically distinct obesity phenotypes in zebrafish. Our results indicate that zebrafish represents an attractive model to study regulatory mechanisms involved in the determination of AT phenotype during development of metabolically healthy versus metabolically unhealthy obesity.
Subject(s)
Adipose Tissue/metabolism , Body Weight/physiology , Diet, High-Fat , Lipid Metabolism/physiology , Obesity/metabolism , Animals , Blood Glucose/metabolism , Disease Models, Animal , Inflammation/metabolism , Male , Triglycerides/metabolism , ZebrafishABSTRACT
BACKGROUND: Nicotinamide phosphoribosyltransferase (NAMPT) and nicotinamide adenine dinucleotide (NAD) levels are crucial for liver function. The saturated fatty acid palmitate and the unsaturated fatty acid oleate are the main free fatty acids in adipose tissue and human diet. We asked how these fatty acids affect cell survival, NAMPT and NAD levels in HepG2 cells and primary human hepatocytes. METHODS: HepG2 cells were stimulated with palmitate (0.5mM), oleate (1mM) or a combination of both (0.5mM/1mM) as well as nicotinamide mononucleotide (NMN) (0.5 mM) or the specific NAMPT inhibitor FK866 (10nM). Cell survival was measured by WST-1 assay and Annexin V/propidium iodide staining. NAD levels were determined by NAD/NADH Assay or HPLC. Protein and mRNA levels were analysed by Western blot analyses and qPCR, respectively. NAMPT enzyme activity was measured using radiolabelled 14C-nicotinamide. Lipids were stained by Oil red O staining. RESULTS: Palmitate significantly reduced cell survival and induced apoptosis at physiological doses. NAMPT activity and NAD levels significantly declined after 48h of palmitate. In addition, NAMPT mRNA expression was enhanced which was associated with increased NAMPT release into the supernatant, while intracellular NAMPT protein levels remained stable. Oleate alone did not influence cell viability and NAMPT activity but ameliorated the negative impact of palmitate on cell survival, NAMPT activity and NAD levels, as well as the increased NAMPT mRNA expression and secretion. NMN was able to normalize intracellular NAD levels but did not ameliorate cell viability after co-stimulation with palmitate. FK866, a specific NAMPT inhibitor did not influence lipid accumulation after oleate-treatment. CONCLUSIONS: Palmitate targets NAMPT activity with a consequent cellular depletion of NAD. Oleate protects from palmitate-induced apoptosis and variation of NAMPT and NAD levels. Palmitate-induced cell stress leads to an increase of NAMPT mRNA and accumulation in the supernatant. However, the proapoptotic action of palmitate seems not to be mediated by decreased NAD levels.
Subject(s)
Cytokines/genetics , Hepatocytes/drug effects , NAD/metabolism , Nicotinamide Phosphoribosyltransferase/genetics , Oleic Acid/pharmacology , Palmitic Acid/pharmacology , Acrylamides/pharmacology , Apoptosis/drug effects , Cell Survival/drug effects , Cytokines/antagonists & inhibitors , Cytokines/metabolism , Enzyme Inhibitors/pharmacology , Gene Expression , Hep G2 Cells , Hepatocytes/cytology , Hepatocytes/metabolism , Humans , NAD/antagonists & inhibitors , Nicotinamide Mononucleotide/pharmacology , Nicotinamide Phosphoribosyltransferase/antagonists & inhibitors , Nicotinamide Phosphoribosyltransferase/metabolism , Palmitic Acid/antagonists & inhibitors , Piperidines/pharmacology , Primary Cell Culture , RNA, Messenger/genetics , RNA, Messenger/metabolismABSTRACT
Patients with phosphatase and tensin homolog (PTEN) hamartoma tumor syndrome and germline mutations in PTEN frequently develop lipomatosis, for which there is no standard treatment. Rapamycin was shown to reduce the growth of lipoma cells with heterozygous PTEN deficiency in vitro, but concomitantly induced an upregulation of AKT phosphorylation. Since it was shown that resveratrol stabilizes PTEN, we asked whether co-incubation with resveratrol could suppress the rapamycin-induced AKT phosphorylation in PTEN-deficient lipoma cells. Resveratrol incubation resulted in decreased lipoma cell viability by inducing G1-phase cell cycle arrest and apoptosis. PTEN expression and AKT phosphorylation were not significantly changed, whereas p70S6 kinase (p70S6K) phosphorylation was reduced in PTEN-deficient lipoma cells after resveratrol incubation. Rapamycin/resveratrol co-incubation significantly decreased viability further at lower doses of resveratrol and resulted in decreased p70S6K phosphorylation compared to rapamycin incubation alone, suggesting that resveratrol potentiated the growth inhibitory effects of rapamycin by reducing p70S6K activation. Both viability and p70S6K phosphorylation of primary PTEN wild-type preadipocytes were less affected compared to PTEN-deficient lipoma cells by equimolar concentrations of resveratrol. These results support the concept of combining chemopreventive natural compounds with mammalian target of rapamycin (mTOR) inhibitors to increase the efficacy of chemotherapeutic drugs for patients suffering from overgrowth syndromes.
Subject(s)
Lipoma/drug therapy , PTEN Phosphohydrolase/metabolism , Ribosomal Protein S6 Kinases, 70-kDa/antagonists & inhibitors , Sirolimus/pharmacology , Stilbenes/pharmacology , Adipocytes/drug effects , Antineoplastic Combined Chemotherapy Protocols/pharmacology , Apoptosis/drug effects , Cell Line, Tumor , G1 Phase Cell Cycle Checkpoints/drug effects , Humans , Lipoma/metabolism , Lipoma/pathology , Phosphorylation/drug effects , Protein Kinase Inhibitors/pharmacology , Resveratrol , Ribosomal Protein S6 Kinases, 70-kDa/metabolism , Sirolimus/administration & dosage , Stilbenes/administration & dosageABSTRACT
BACKGROUND: Nicotinamide phosphoribosyltransferase (NAMPT) is the key enzyme of the NAD salvage pathway starting from nicotinamide. Cancer cells have an increased demand for NAD due to their high proliferation and DNA repair rate. Consequently, NAMPT is considered as a putative target for anti-cancer therapies. There is evidence that AMP-activated protein kinase (AMPK) and mammalian target of rapamycin (mTOR) become dysregulated during the development of hepatocellular carcinoma (HCC). Here, we investigated the effects of NAMPT inhibition by its specific inhibitor FK866 on the viability of hepatocarcinoma cells and analyzed the effects of FK866 on the nutrient sensor AMPK and mTOR complex1 (mTORC1) signaling. RESULTS: FK866 markedly decreased NAMPT activity and NAD content in hepatocarcinoma cells (Huh7 cells, Hep3B cells) and led to delayed ATP reduction which was associated with increased cell death. These effects could be abrogated by administration of nicotinamide mononucleotide (NMN), the enzyme product of NAMPT. Our results demonstrated a dysregulation of the AMPK/mTOR pathway in hepatocarcinoma cells compared to non-cancerous hepatocytes with a higher expression of mTOR and a lower AMPKα activation in hepatocarcinoma cells. We found that NAMPT inhibition by FK866 significantly activated AMPKα and inhibited the activation of mTOR and its downstream targets p70S6 kinase and 4E-BP1 in hepatocarcinoma cells. Non-cancerous hepatocytes were less sensitive to FK866 and did not show changes in AMPK/mTOR signaling after FK866 treatment. CONCLUSION: Taken together, these findings reveal an important role of the NAMPT-mediated NAD salvage pathway in the energy homeostasis of hepatocarcinoma cells and suggest NAMPT inhibition as a potential treatment option for HCC.
Subject(s)
AMP-Activated Protein Kinases/metabolism , Acrylamides/pharmacology , Carcinoma, Hepatocellular/metabolism , Cytokines/antagonists & inhibitors , Cytokines/metabolism , Liver Neoplasms/metabolism , Nicotinamide Phosphoribosyltransferase/antagonists & inhibitors , Nicotinamide Phosphoribosyltransferase/metabolism , Piperidines/pharmacology , TOR Serine-Threonine Kinases/metabolism , Adenosine Triphosphate/metabolism , Apoptosis/drug effects , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/pathology , Cell Line, Tumor , Cell Survival/drug effects , Down-Regulation/drug effects , Enzyme Activation , Gene Expression Regulation, Neoplastic , Humans , Liver Neoplasms/genetics , Liver Neoplasms/pathology , Signal Transduction/drug effectsABSTRACT
BACKGROUND: Phosphatase and tensin homolog (PTEN) hamartoma tumor syndrome (PHTS) is caused by germ line mutations in the PTEN gene. Symptoms include cancer predisposition, immune deviations, and lipomas/lipomatosis. No causal standard therapy is available. We describe a therapeutic attempt with the mammalian target of rapamycin (mTOR) inhibitor sirolimus for a PHTS patient suffering from thymus hyperplasia and lipomatosis. We furthermore assessed the in vitro effects of sirolimus and other inhibitors on lipoma cells of the patient. METHODS: The patient underwent clinical and blood examinations and whole-body magnetic resonance imaging to assess tumor sizes. Lipoma cells of the patient were incubated with inhibitors of the phosphoinositide-3-kinase (PI3K)/AKT/mTOR signaling pathway to analyze the effects on proliferation, adipocyte differentiation, and survival in vitro. RESULTS: Sirolimus treatment improved somatic growth and reduced thymus volume. These effects diminished over the treatment period of 19 mo. Sirolimus decreased lipoma cell proliferation and adipocyte differentiation in vitro but did not cause apoptosis. PI3K and AKT inhibitors induced apoptosis significantly. CONCLUSION: Sirolimus treatment led to an improvement of the patient's clinical status and a transient reduction of the thymus. Our in vitro findings point to PI3K and AKT inhibitors as potential treatment options for patients with severe forms of PHTS.
Subject(s)
Hamartoma Syndrome, Multiple/drug therapy , PTEN Phosphohydrolase/genetics , Sirolimus/therapeutic use , Child, Preschool , Hamartoma Syndrome, Multiple/genetics , Humans , In Vitro Techniques , Infant , Infant, NewbornABSTRACT
The adipokine leptin has been detected in human breast milk, but its effect on postnatal growth and development remains largely unclear. We hypothesized that leptin could affect infant's body weight gain during early lactation in the first 6 mo of life. Therefore, we evaluated leptin levels in maternal serum and breast milk of 23 healthy, lactating mothers and their neonates in a prospective, longitudinal study. Leptin concentration was quantified by a commercially available human leptin RIA. Our results showed that leptin levels in breast milk were 22-fold lower than in maternal serum, but both parameters were positively correlated to each other (r = 0.431, p = 0.001) and to maternal BMI (serum: r = 0.512, p < 0.001; milk: r = 0.298, p < 0.001) over 6 mo of lactation. A negative association was found between breast milk leptin levels during the first week after delivery and the infant weight gain from the end of the first to the sixth month (r = -0.681, p = 0.007). This suggests that milk-borne leptin provides a link between maternal body composition and infant growth and development and plays a critical role in regulating appetite and food intake during early infancy.
Subject(s)
Child Development/physiology , Lactation/physiology , Leptin/analysis , Milk, Human/chemistry , Weight Gain/physiology , Adult , Body Composition/physiology , Female , Humans , Infant , Infant, Newborn , Leptin/blood , Longitudinal Studies , Radioimmunoassay , Statistics, NonparametricABSTRACT
Nicotinamide phosphoribosyltransferase Nicotinamide phosphoribosyltransferase (Nampt Nampt ) is a key nicotinamide adenine dinucleotide (NAD) NAD biosynthetic enzyme in mammals, converting nicotinamide nicotinamide into nicotinamide mononucleotide nicotinamide mononucleotide (NMN NMN ), an NAD intermediate. First identified in humans as a cytokine cytokine pre-B-cell colony enhancing factor pre-B cell colony enhancing factor (PBEF PBEF ) and subsequently described as an insulin-mimetic hormone visfatin visfatin , Nampt has recently excited the scientific interest of researchers from diverse fields, including NAD biology, metabolic regulation, and inflammation. As an NAD biosynthetic enzyme, Nampt regulates the activity of NAD-consuming enzymes such as sirtuins sirtuins and influences a variety of metabolic and stress responses. Nampt plays an important role in the regulation of insulin secretion insulin secretion in pancreatic ß-cells. Nampt also functions as an immunomodulatory cytokine cytokine and is involved in the regulation of inflammatory responses. This chapter summarizes the various functional aspects of Nampt and discusses its potential roles in diseases, with special focus on type 2 diabetes mellitus (T2DM).
Subject(s)
Diabetes Mellitus, Type 2/enzymology , Diabetes Mellitus, Type 2/physiopathology , Nicotinamide Phosphoribosyltransferase/physiology , Animals , Biomarkers , Diabetes Mellitus, Type 2/blood , Humans , Inflammation/enzymology , Inflammation/pathology , Insulin-Secreting Cells/enzymology , Insulin-Secreting Cells/physiology , Liver/enzymology , Nicotinamide Phosphoribosyltransferase/blood , Nicotinamide Phosphoribosyltransferase/metabolism , Obesity/bloodABSTRACT
INTRODUCTION: In emergency care, geriatric requirements and risks are often not taken sufficiently into account. In addition, there are neither evidence-based recommendations nor scientifically developed quality indicators (QI) for geriatric emergency care in German emergency departments. As part of the GeriQ-ED© research project, quality indicators for geriatric emergency medicine in Germany have been developed using the QUALIFY-instruments. METHODS: Using a triangulation methodology, a) clinical experience-based quality aspects were identified and verified, b) research-based quality statements were formulated and assessed for relevance, and c) preliminary quality indicators were operationalized and evaluated in order to recommend a feasible set of final quality indicators. RESULTS: Initially, 41 quality statements were identified and assessed as relevant. Sixty-seven QI (33 process, 29 structure and 5 outcome indicators) were extrapolated and operationalised. In order to facilitate implementation into daily practice, the following five quality statements were defined as the GeriQ-ED© TOP 5: screening for delirium, taking a full medications history including an assessment of the indications, education of geriatric knowledge and skills to emergency staff, screening for patients with geriatric needs, and identification of patients with risk of falls/ recurrent falls. DISCUSSION: QIs are regarded as gold standard to measure, benchmark and improve emergency care. GeriQ-ED© QI focused on clinical experience- and research-based recommendations and describe for the first time a standard for geriatric emergency care in Germany. GeriQ-ED© TOP 5 should be implemented as a minimum standard in geriatric emergency care.
Subject(s)
Emergency Medicine/standards , Emergency Service, Hospital , Geriatric Assessment , Geriatrics/standards , Quality Indicators, Health Care , Accidental Falls/prevention & control , Aged , Delirium/diagnosis , Delphi Technique , Humans , Inservice Training , Mass Screening/standards , Medication Reconciliation , Needs Assessment , Quality Improvement , Risk AssessmentABSTRACT
LDL-cholesterol (LDL-C) is a causal pathogenic factor in atherosclerosis. Monoclonal anti-proprotein convertase subtilisin/kexin type 9 (PCSK9) neutralizing antibodies are novel potent LDL-lowering drugs which reduce cardiovascular events. To characterize their effect on atherogenesis, APOE*3Leiden.CETP mice were fed a high cholesterol/high fat diet (WTD) or normal chow (NC) for 18 weeks. Mice on WTD were injected with the human anti-PCSK9 antibody mAb1 (PL-45134, 10 mg*kg-1 s.c.) or 0.9% saline every 10 days. PCSK9 inhibition decreased total cholesterol in serum of APOE*3Leiden.CETP mice and prevented the development of atherosclerosis. The plaque area in the aortic root was reduced by half and macrophage infiltration determined by Ly6c and Mac-3 staining was ameliorated. PCSK9 inhibition decreased markers of inflammation in mononuclear cells (Il-6, Tnfa mRNA), and in serum (CXCL-1,-10,-13; complement factor C5a) compared to control WTD fed animals. The number of circulating Sca-1/VEGF-R2 positive endothelial progenitor cells of the peripheral blood and spleen-derived diLDL/lectin double positive circulating angiogenic cells was increased. To conclude, the PCSK9-mediated anti-atherosclerotic effect involves the upregulation of pro-regeneratory endothelial progenitor cells, a reduction of inflammation and change of plaque composition.
Subject(s)
Apolipoproteins E/metabolism , Atherosclerosis/metabolism , Plaque, Atherosclerotic/metabolism , Proprotein Convertase 9/metabolism , Animals , Antibodies, Monoclonal/physiology , Atherosclerosis/drug therapy , Cholesterol/metabolism , Cholesterol, LDL/metabolism , Endothelial Progenitor Cells/drug effects , Endothelial Progenitor Cells/metabolism , Humans , Hypolipidemic Agents/pharmacology , Inflammation/metabolism , Lectins/metabolism , Macrophages/drug effects , Macrophages/metabolism , Mice , Plaque, Atherosclerotic/drug therapy , Up-Regulation/drug effectsABSTRACT
Nonalcoholic steatohepatitis (NASH) is considered the progressive form of nonalcoholic fatty liver disease (NAFLD) and is characterized by liver steatosis, inflammation, hepatocellular injury and different degrees of fibrosis. A central issue in this field relates to the identification of those factors that trigger inflammation, thus fuelling the transition from nonalcoholic fatty liver to NASH. These triggers of liver inflammation might have their origins outside the liver (such as in adipose tissue or the gut) as well as inside the organ (for instance, lipotoxicity, innate immune responses, cell death pathways, mitochondrial dysfunction and endoplasmic reticulum stress), both of which contribute to NASH development. In this Review, we summarize the currently available information on the key upstream triggers of inflammation in NASH. We further delineate the mechanisms by which liver inflammation is resolved and the implications of a defective pro-resolution process. A better knowledge of these mechanisms should help to design targeted therapies able to halt or reverse disease progression.
Subject(s)
Non-alcoholic Fatty Liver Disease/etiology , Humans , Inflammation/etiology , Inflammation/pathology , Liver/pathology , Non-alcoholic Fatty Liver Disease/pathology , Non-alcoholic Fatty Liver Disease/therapyABSTRACT
NAMPT (Nicotinamide phosphoribosyltransferase) catalyses the rate-limiting step in the NAD biosynthesis from nicotinamide and thereby regulates the activity of NAD-dependent enzymes. Cancer cells are highly dependent on NAD for energy and DNA repair processes and are assumed to be more susceptible to an inhibition of NAD synthesis than non-transformed cells. We aimed to investigate whether or not inhibition of NAMPT with its specific inhibitor FK866 can sensitize leukemia cells for chemotherapeutic agents. NAMPT protein abundance, enzymatic activity and NAD concentrations were significantly higher in Jurkat and Molt-4 leukemia cell lines compared to normal peripheral blood mononuclear cells. Combination of etoposide and FK866 caused increased cell death in leukemia cell lines compared to etoposide alone. Etoposide decreased protein abundance of NAD-dependent deacetylases SIRTUIN1. After combining etoposide and FK866 treatment SIRTUIN2 was further decreased and accumulation and acetylation of the downstream target p53 was further enhanced in MOLT4 cells. Concomitantly, protein abundance of p21 and cleaved BAX was increased. Targeting NAMPT could be a novel therapeutic strategy to enhance the efficacy of chemotherapeutic agents such as etoposide against leukemia.
Subject(s)
Cytokines/antagonists & inhibitors , Enzyme Inhibitors/pharmacology , Etoposide/pharmacology , Leukemia/drug therapy , Nicotinamide Phosphoribosyltransferase/antagonists & inhibitors , Sirtuin 2/metabolism , Topoisomerase II Inhibitors/pharmacology , Tumor Suppressor Protein p53/metabolism , Cell Death/drug effects , Cell Line, Tumor , Humans , Leukemia/enzymology , Leukemia/metabolism , Signal TransductionABSTRACT
Background Adipokines were shown to affect glucose homeostasis and ß-cell function in patients with pancreatic dysfunction which is associated with changes in the adipose tissue secretory profile. However, information about adipokines associated with ß-cell dysfunction is lacking in pediatric patients with type 1 diabetes. Methods (1) We compared serum concentrations of nicotinamide phosphoribosyltransferase (NAMPT), omentin-1 and caspase-cleaved cytokeratin 18 fragment M30 (CK-18) in pediatric type 1 diabetes patients (n=245) and healthy age, sex and body mass index standard deviation score (BMI-SDS) matched controls (n=243). (2) We investigated the influence of insulin treatment on serum concentrations of NAMPT, omentin-1 and CK-18 in groups of patients with type 1 diabetes stratified according to the duration of their disease: at onset (n=50), ≥6 months and <5 years (n=185), ≥5 and <10 years (n=98), and ≥10 years (n=52). Results Patients at onset compared with healthy controls demonstrated no significant differences in NAMPT levels (p=0.129), whereas omentin-1 levels were elevated (p<0.001) and CK-18 levels were lowered (p=0.034). In contrast, NAMPT and omentin-1 were elevated and CK-18 serum levels were lower in longstanding patients compared to healthy controls (p<0.001). NAMPT serum levels did not change significantly during the duration of type 1 diabetes (p=0.546). At onset, omentin-1 and CK-18 levels were higher than in any group of longstanding type 1 diabetes (p<0.025). Conclusions Altered serum levels of NAMPT, omentin-1 and CK-18 in pediatric type 1 diabetes patients indicate metabolic changes caused by adipose tissue dysregulation which do not normalize during insulin therapy.
Subject(s)
Cytokines/blood , Diabetes Mellitus, Type 1/blood , Keratin-18/blood , Lectins/blood , Nicotinamide Phosphoribosyltransferase/blood , Adolescent , Blood Glucose , Body Mass Index , Child , Female , GPI-Linked Proteins/blood , Humans , Insulin/blood , Insulin Resistance/physiology , MaleABSTRACT
BACKGROUND: Linoleic acid (LA) is abundant in modern industrialized diets. Oxidized LA metabolites (OXLAMs) and reactive aldehydes, such as 4-hydroxy-2-nonenal (4-HNE), are present in heated vegetable oils and can be endogenously synthesized following consumption of dietary LA. OXLAMs have been implicated in cerebellar degeneration in chicks; 4-HNE is linked to neurodegenerative conditions in mammals. It unknown whether increasing dietary LA or OXLAMs alters the levels of oxidized fatty acids (oxylipins), precursor fatty acids, or 4-HNE in mammalian brain. OBJECTIVES: To determine the effects of increases in dietary OXLAMs and dietary LA, on levels of fatty acids, oxylipins, and 4-HNE in mouse brain tissues. METHODS: Mice (nâ¯=â¯8 per group) were fed one of three controlled diets for 8â¯weeks: (1) a low LA diet, (2) a high LA diet, or (3) the low LA diet with added OXLAMs. Brain fatty acids, oxylipins, and 4-HNE were quantified in mouse cerebellum and cerebral cortex by gas chromatography-flame ionization detection, liquid chromatography-tandem mass spectrometry, and immunoblot, respectively. RESULTS: Increasing dietary LA significantly increased omega-6 fatty acids, decreased omega-3 fatty acids, and increased OXLAMs in brain. Dietary OXLAMs had minimal effect on oxidized lipids but did decrease both omega-6 and omega-3 fatty acids. Neither dietary LA nor OXLAMs altered 4-HNE levels. CONCLUSION: Brain fatty acids are modulated by both dietary LA and OXLAMs, while brain OXLAMs are regulated by endogenous synthesis from LA, rather than incorporation of preformed OXLAMs.
Subject(s)
Aldehydes/metabolism , Brain/metabolism , Diet , Linoleic Acids/pharmacology , Lipid Peroxidation , Oxylipins/metabolism , Animals , Male , Metabolome , Mice, Inbred C57BL , Oxidation-ReductionABSTRACT
Nicotinamide phosphoribosyltransferase (NAMPT) is a regulator of the intracellular nicotinamide adenine dinucleotide (NAD) pool. NAD is an essential coenzyme involved in cellular redox reactions and is a substrate for NAD-dependent enzymes. In various metabolic disorders and during ageing, levels of NAD are decreased. Through its NAD-biosynthetic activity, NAMPT influences the activity of NAD-dependent enzymes, thereby regulating cellular metabolism. In addition to its enzymatic function, extracellular NAMPT (eNAMPT) has cytokine-like activity. Abnormal levels of eNAMPT are associated with various metabolic disorders. NAMPT is able to modulate processes involved in the pathogenesis of obesity and related disorders such as nonalcoholic fatty liver disease (NAFLD) and type 2 diabetes mellitus (T2DM) by influencing the oxidative stress response, apoptosis, lipid and glucose metabolism, inflammation and insulin resistance. NAMPT also has a crucial role in cancer cell metabolism, is often overexpressed in tumour tissues and is an experimental target for antitumour therapies. In this Review, we discuss current understanding of the functions of NAMPT and highlight progress made in identifying the physiological role of NAMPT and its relevance in various human diseases and conditions, such as obesity, NAFLD, T2DM, cancer and ageing.