RESUMO
Harvest season exerts great influence on tea quality. Herein, the variations in non-volatile flavor substances in spring and summer fresh tea leaves of four varieties were comprehensively investigated by integrating UHPLC-Q-Exactive based lipidomics and metabolomics. A total of 327 lipids and 99 metabolites were detected, among which, 221 and 58 molecules were significantly differential. The molecular species of phospholipids, glycolipids and acylglycerolipids showed most prominent and structure-dependent seasonal changes, relating to polar head, unsaturation and total acyl length. Particularly, spring tea contained higher amount in aroma precursors of highly unsaturated glycolipids and phosphatidic acids. The contents of umami-enhancing amino acids and phenolic acids, e.g., theanine, theogallin and gallotannins, were increased in spring. Besides, catechins, theaflavins, theasinensins and flavone/flavonol glycosides showed diverse changes. These phytochemical differences covered key aroma precursors, tastants and colorants, and may confer superior flavor of black tea processed using spring leaves, which was verified by sensory evaluation.
Assuntos
Camellia sinensis , Aromatizantes , Lipidômica , Espectrometria de Massas , Metabolômica , Folhas de Planta , Estações do Ano , Camellia sinensis/química , Camellia sinensis/metabolismo , Folhas de Planta/química , Folhas de Planta/metabolismo , Cromatografia Líquida de Alta Pressão , Aromatizantes/química , Aromatizantes/metabolismo , Humanos , Paladar , Odorantes/análise , Lipídeos/análise , Lipídeos/químicaRESUMO
Reversed-phase ultrahigh-performance liquid chromatography-mass spectrometry (RP-UHPLC/MS) method is optimized for the quantitation of a large number of lipid species in biological samples, primarily in human plasma and serum. The method uses a C18 bridged ethylene hybrid (BEH) column (150 × 2.1 mm; 1.7 µm) for the separation of lipids from 23 subclasses with a total run time of 25 min. Lipid species separation allows the resolution of isobaric and isomeric lipid forms. A triple quadrupole mass spectrometer is used for targeted lipidomic analysis using multiple reaction monitoring (MRM) in the positive ion mode. Data are evaluated by Skyline software, and the concentrations of analytes are determined using internal standards per each individual lipid class.
Assuntos
Cromatografia de Fase Reversa , Lipidômica , Lipídeos , Cromatografia Líquida de Alta Pressão/métodos , Cromatografia de Fase Reversa/métodos , Humanos , Lipidômica/métodos , Lipídeos/análise , Espectrometria de Massas/métodos , Ensaios de Triagem em Larga Escala/métodos , Espectrometria de Massas em Tandem/métodos , Software , Espectrometria de Massa com Cromatografia LíquidaRESUMO
Sphingolipids (SLs) are essential lipids with important functions in membrane formation and cell signaling. The presence of a long chain base (LCB) structure is common to all SLs. De novo SL synthesis is initiated by the enzyme serine-palmitoyltransferase (SPT), which forms an LCB by the conjugation from serine and fatty acyl-CoAs. SPT can metabolize a variety of acyl-CoA substrates, which form diverse LCB structures within and across species. The LCB then undergoes further metabolic modifications resulting in an extraordinarily diverse spectrum of sphingolipids formed. SL analysis, using liquid chromatography-mass spectrometry (LC-MS)-based methods, poses challenges due to the diverse range of frequently isobaric species. This complexity complicates the identification of underlying LCB structures using standard lipidomics approaches. Here, we describe a simplified method to analyze the LCB profile in cells, tissue, and blood. The procedure involves chemical hydrolysis to remove the conjugated headgroups and N-acyl chains, allowing to specifically resolve the underlying LCB structures by LC-MS. This method can also be combined with an isotope labeling approach to determine in vivo SPT activity and total SL de novo synthesis over time.
Assuntos
Esfingolipídeos , Cromatografia Líquida/métodos , Esfingolipídeos/metabolismo , Esfingolipídeos/análise , Esfingolipídeos/química , Lipidômica/métodos , Espectrometria de Massas/métodos , Animais , Humanos , Serina C-Palmitoiltransferase/metabolismo , Acil Coenzima A/metabolismo , Espectrometria de Massas em Tandem/métodosRESUMO
Sphingolipids are the most diverse class of lipids due to the numerous variations in their structural components. This diversity is also reflected in their extremely different functions. Sphingolipids are not only constituents of cell membranes but have emerged as key signaling molecules involved in a variety of cellular functions, such as cell growth and differentiation, proliferation and apoptotic cell death. Lipidomic analyses in clinical research have identified pathways and products of sphingolipid metabolism that are altered in several human pathologies. In this article, we describe how to properly design a lipidomic experiment in clinical research, how to handle plasma and serum samples for this purpose, and how to measure sphingolipids using liquid chromatography-mass spectrometry.
Assuntos
Lipidômica , Esfingolipídeos , Esfingolipídeos/metabolismo , Esfingolipídeos/sangue , Humanos , Lipidômica/métodos , Cromatografia Líquida/métodos , Espectrometria de Massas/métodosRESUMO
Bioactive lipid mediators derived from arachidonic acid constitute an attractive pool of metabolites that reflect cellular function and signaling, as well as potential biomarkers that may respond quantitatively to disease progression or pharmacological treatment. Their quantitative measurement in biological samples is complicated by the number of isomers that share common structural features, which are not easily distinguished by immunoassays or reverse phase chromatography-tandem mass spectrometry. Here, we present a method that enables the rapid analysis of a panel of over 25 biologically important eicosanoids in a 96-well format for cell culture supernatants, plasma, and organ tissues using convergence chromatography-tandem mass spectrometry to resolve these analytes of interest.
Assuntos
Espectrometria de Massas em Tandem , Espectrometria de Massas em Tandem/métodos , Humanos , Eicosanoides/análise , Eicosanoides/metabolismo , Animais , Cromatografia Líquida/métodos , Lipídeos/análise , Lipídeos/química , Biomarcadores , Lipidômica/métodosRESUMO
Shotgun Lipidomics is a robust methodology for the characterization of the lipidome of complex biological samples. This assay is among the most quantitative lipidomics methods and is capable of surveying a wide breadth of lipid subclasses, both neutral and polar. The shortfalls of the technique include limitations in lipid species characterization and computationally demanding data analysis requiring isotopic and isobaric overlap correction. Differential Mobility Spectrometry (DMS) has demonstrated its utility in enabling acyl tail characterization within a Shotgun Lipidomics experiment. Here, we present a workflow for DMS Shotgun Lipidomics that measures 1400 possible lipid species. It utilizes the Shotgun Lipidomics Assistant (SLA) application, an open-source application that supervises the data analysis for an expansive Shotgun Lipidomics experiment.
Assuntos
Lipidômica , Lipídeos , Lipidômica/métodos , Animais , Lipídeos/análise , Lipídeos/química , Software , Fluxo de Trabalho , Mamíferos/metabolismo , Metabolismo dos Lipídeos , Espectrometria de Mobilidade Iônica/métodos , Humanos , Espectrometria de Massas/métodosRESUMO
Recent developments in LC-MS instrumentation and analytical technologies together with bioinformatics tools supporting high-throughput processing of large omics datasets significantly enhanced our capabilities and efficiency of identification and quantification of lipids in diverse biological materials. However, each biological matrix is characterized by its unique lipid composition, thus requiring optimization of analytical and bioinformatics workflows for each studied lipidome. Here, we describe an integrated workflow for deep lipidome profiling, accurate annotation, and semi-absolute quantification of complex lipidomes based on reversed phase chromatography and high resolution mass spectrometry. This chapter provides details on selection of the optimal extraction protocol, acquisition of LC-MS/MS data for accurate annotation of lipid molecular species, and design of lipidome-specific mixtures of internal standards to assist quantitative analysis of complex, native lipidomes.
Assuntos
Lipidômica , Lipídeos , Espectrometria de Massas em Tandem , Espectrometria de Massas em Tandem/métodos , Lipidômica/métodos , Lipídeos/análise , Lipídeos/química , Cromatografia Líquida/métodos , Biologia Computacional/métodos , Fluxo de Trabalho , Humanos , Cromatografia de Fase Reversa/métodos , Espectrometria de Massa com Cromatografia LíquidaRESUMO
Ultrahigh-performance supercritical fluid chromatography-mass spectrometry (UHPSFC/MS) method is optimized for the high-throughput quantitation of lipids in human serum and plasma with an emphasis on robustness and accurate quantitation. Bridged ethylene hybrid (BEH) silica column (100 × 3 mm; 1.7 µm) is used for the separation of 17 nonpolar and polar lipid classes in 4.4 min using the positive ion electrospray ionization mode. The lipid class separation approach in UHPSFC/MS results in the coelution of all lipid species within one lipid class in one chromatographic peak, including two exogenous internal standards (IS) per lipid class, which provides the optimal conditions for robust quantitation. The method was validated according to European Medicines Agency and Food and Drug Administration recommendations. UHPSFC/MS combined with LipidQuant software allows a semiautomated process to determine lipid concentrations with a total run time of only 8 min including column equilibration, which enables the analysis of 160 samples per day.
Assuntos
Cromatografia com Fluido Supercrítico , Lipidômica , Lipídeos , Cromatografia com Fluido Supercrítico/métodos , Humanos , Lipidômica/métodos , Lipídeos/análise , Lipídeos/sangue , Espectrometria de Massas/métodos , Cromatografia Líquida de Alta Pressão/métodosRESUMO
Cardiolipins (CL) are special lipids in many respects. First of all, CL are composed of four fatty acids linked by two phosphatidic acids, which provide CL a unique molecular structure. Secondly, in eukaryotic cells they are specific to a single organelle, mitochondria, where they are also synthetized. CL are one of the most abundant lipid classes in mitochondria, mainly localized in the inner membrane. They are key determinants of mitochondrial health and homeostasis by modulating membrane integrity and fluidity, mitochondrial shapes, and metabolic pathways. Disturbances in mitochondrial CL composition can lead to tissue malfunction and diseases. It is therefore important to develop analytical tools to study the mitochondrial lipidome, and more particularly the CL. The method described here allows the quantification of cardiolipins at the sum composition level in isolated mitochondria or in liver tissue by flow injection analysis coupled to differential mobility spectrometry (FIA-DMS), also known as DMS-based shotgun lipidomics.
Assuntos
Cardiolipinas , Lipidômica , Cardiolipinas/análise , Cardiolipinas/metabolismo , Lipidômica/métodos , Animais , Mitocôndrias/metabolismo , Espectrometria de Massas/métodos , Fígado/metabolismo , Fígado/químicaRESUMO
Cell cultures are widely used in studies to gain mechanistic insights of metabolic processes. The foundation of these studies lies on the quantification of intracellular and extracellular metabolites, and nuclear magnetic resonance (NMR) is one of the key analytical platforms used to this aim. Among the factors influencing the quality of the produced data are the sampling procedures as well as the acquisition and processing of spectroscopic data. Here we provide our workflow for obtaining quantitative metabolic data from adherent mammalian cells using NMR spectroscopy. The described protocol is compatible with other analytical methods like LC- or GC-MS-based lipidomics and untargeted metabolomics from the same sample. We also show how the collected extracellular data can be used to extract exchange flux rates, particularly useful for flux analysis studies and metabolic engineering of human-induced pluripotent stem cells.
Assuntos
Metabolismo Energético , Espectroscopia de Ressonância Magnética , Metabolômica , Humanos , Metabolômica/métodos , Espectroscopia de Ressonância Magnética/métodos , Células-Tronco Pluripotentes Induzidas/metabolismo , Células-Tronco Pluripotentes Induzidas/citologia , Metaboloma , Animais , Lipidômica/métodosRESUMO
Background: Acne vulgaris (AV), a chronic inflammatory pilosebaceous disorder, affects 80-90% of teenagers. This study aimed to discover lipid profiles and biomarkers of the rabbit ear acne model, and investigate the mechanism of isotretinoin in treating acne at the lipid level. Methods: Untargeted lipidomic analysis using ultra-high performance liquid chromatography system (UHPLC) coupled to q-extraction plus was performed to identify skin lipid metabolites in blank control (groups C), model group (group M) and isotretinoin group (group T). Multivariate statistical analysis was used to process the lipidomics data. Results: A total of 43 lipid classes comprising 6989 lipid species were identified from the mass spectrometry data. The orthogonal partial least squares discriminant analysis (OPLS-DA) model demonstrated significant separation in skin lipidomic profiles between group M and group C. With variable influence on projection (VIP) > 1.0 and P-value < 0.05, 299 significantly different lipid metabolites were identified. These lipid metabolites consisted mainly of ceramides (Cer) (53.85%), phosphatidylethanolamines (PE) (9.03%), phosphatidylcholines (PC)(5.35%), and sphingomyelin (SM)(4.01%). Combining with AUC ≥ 0.9 as the elected criteria, Cer (d18;1_24:0), zymosterol (ZyE)(33:5), Cer (t43:1), ZyE (33:6), ZyE (24:7), and ZyE (35:6) have "high" accuracy. Isotretinoin treatment normalized 25 lipid metabolites in the acne model. Conclusion: Our findings provide new insights into the role of lipid metabolism in the pathogenesis of acne and the action mechanism of isotretinoin.
Assuntos
Acne Vulgar , Biomarcadores , Modelos Animais de Doenças , Isotretinoína , Lipidômica , Lipídeos , Isotretinoína/farmacologia , Acne Vulgar/tratamento farmacológico , Acne Vulgar/metabolismo , Animais , Coelhos , Biomarcadores/metabolismo , Biomarcadores/análise , Lipídeos/análise , Cromatografia Líquida de Alta Pressão , Masculino , Fármacos Dermatológicos/farmacologia , Fármacos Dermatológicos/uso terapêuticoRESUMO
BACKGROUND: Observational studies have revealed associations between maternal lipid metabolites and gestational diabetes mellitus (GDM). However, whether these associations are causal remain uncertain. OBJECTIVE: To evaluate the causal relationship between lipid metabolites and GDM. METHODS: A two-sample Mendelian randomization (MR) analysis was performed based on summary statistics. Sensitivity analyses, validation analyses and reverse MR analyses were conducted to assess the robustness of the MR results. Additionally, a phenome-wide MR (Phe-MR) analysis was performed to evaluate potential side effects of the targeted lipid metabolites. RESULTS: A total of 295 lipid metabolites were included in this study, 29 of them had three or more instrumental variables (IVs) suitable for sensitivity analyses. The ratio of triglycerides to phosphoglycerides (TG_by_PG) was identified as a potential causal biomarker for GDM (inverse variance weighted (IVW) estimate: odds ratio (OR) = 2.147, 95% confidential interval (95% CI) 1.415-3.257, P = 3.26e-4), which was confirmed by validation and reverse MR results. Two other lipid metabolites, palmitoyl sphingomyelin (d18:1/16:0) (PSM(d18:1/16:0)) (IVW estimate: OR = 0.747, 95% CI 0.583-0.956, P = 0.021) and triglycerides in very small very low-density lipoprotein (XS_VLDL_TG) (IVW estimate: OR = 2.948, 95% CI 1.197-5.215, P = 0.015), were identified as suggestive potential biomarkers for GDM using a conventional cut-off P-value of 0.05. Phe-MR results indicated that lowering TG_by_PG had detrimental effects on two diseases but advantageous effects on the other 13 diseases. CONCLUSION: Genetically predicted elevated TG_by_PG are causally associated with an increased risk of GDM. Side-effect profiles indicate that TG_by_PG might be a target for GDM prevention, though caution is advised due to potential adverse effects on other conditions.
Assuntos
Biomarcadores , Diabetes Gestacional , Lipidômica , Lipídeos , Análise da Randomização Mendeliana , Humanos , Diabetes Gestacional/sangue , Diabetes Gestacional/diagnóstico , Diabetes Gestacional/genética , Feminino , Gravidez , Fatores de Risco , Lipídeos/sangue , Medição de Risco , Biomarcadores/sangue , Fenótipo , Predisposição Genética para Doença , Reprodutibilidade dos Testes , FenômicaRESUMO
Elevated triglycerides (TG) are a risk factor for cardiometabolic disorders. There are limited data on lipidomics profiles associated with serum triglycerides concentrations, although these could advance our understanding of the mechanisms underlying these associations. We conducted a lipidomics study of 308 Nigerians with replication in 199 Kenyans. Regression models were used to assess the association of TG with 480 lipid metabolites. Association and mediation analyses were conducted to determine the relationship among TG, metabolites, and several cardiometabolic traits. Ninety-nine metabolites were significantly associated with TG, and 91% of these associations replicated. Overrepresentation analysis identified enrichment of diacylglycerols, monoacylglycerols, diacylglycerophosphoethanolamines, monoacylglycerophosphocholines, ceramide phosphocholines, and diacylglycerophosphocholines. TG-cardiometabolic trait associations were largely mediated by TG-associated metabolites. Associations with type 2 diabetes, waist circumference, body mass index, total cholesterol, and low-density lipoprotein cholesterol concentration were independently mediated by metabolites in multiple subpathways. This lipidomics study in sub-Saharan Africans demonstrated that TG is associated with several non-TG lipids classes, including phosphatidylethanolamines, phosphatidylcholines, lysophospholipids, and plasmalogens, some of which may mediate the effect of TG as a risk factor for cardiometabolic disorders. The study identifies metabolites that are more proximal to cardiometabolic traits, which may be useful for understanding the underlying biology as well as differences in TG-trait associations across ancestries.
Assuntos
Lipidômica , Triglicerídeos , Humanos , Triglicerídeos/sangue , Lipidômica/métodos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/metabolismo , Índice de Massa Corporal , Circunferência da Cintura , África SubsaarianaRESUMO
BACKGROUND: Atmospheric particulate matter (PM) exposure-induced neuroinflammation is critical in mediating nervous system impairment. However, effective intervention is yet to be developed. RESULTS: In this study, we examine the effect of ß-nicotinamide mononucleotide (NMN) supplementation on nervous system damage upon PM exposure and the mechanism of spatial regulation of lipid metabolism. 120 C57BL/6 male mice were exposed to real ambient PM for 11 days (subacute) or 16 weeks (sub-chronic). NMN supplementation boosted the level of nicotinamide adenine dinucleotide (NAD+) in the mouse brain by 2.04 times. This augmentation effectively reduced neuroinflammation, as evidenced by a marked decrease in activated microglia levels across various brain regions, ranging from 29.29 to 85.96%. Whole brain lipidomics analysis revealed that NMN intervention resulted in an less increased levels of ceramide (Cer) and lysophospholipid in the brain following subacute PM exposure, and reversed triglyceride (TG) and glycerophospholipids (GP) following sub-chronic PM exposure, which conferred mice with anti-neuroinflammation response, improved immune function, and enhanced membrane stability. In addition, we demonstrated that the hippocampus and hypothalamus might be the most sensitive brain regions in response to PM exposure and NMN supplementation. Particularly, the alteration of TG (60:10, 56:2, 60:7), diacylglycerol (DG, 42:6), and lysophosphatidylcholine (LPC, 18:3) are the most profound, which correlated with the changes in functional annotation and perturbation of pathways including oxidative stress, inflammation, and membrane instability unveiled by spatial transcriptomic analysis. CONCLUSIONS: This study demonstrates that NMN intervention effectively reduces neuroinflammation in the hippocampus and hypothalamus after PM exposure by modulating spatial lipid metabolism. Strategies targeting the improvement of lipid homeostasis may provide significant protection against brain injury associated with air pollutant exposure.
Assuntos
Encéfalo , Metabolismo dos Lipídeos , Camundongos Endogâmicos C57BL , Material Particulado , Animais , Metabolismo dos Lipídeos/efeitos dos fármacos , Masculino , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Material Particulado/toxicidade , Camundongos , Doenças Neuroinflamatórias/induzido quimicamente , Doenças Neuroinflamatórias/metabolismo , Suplementos Nutricionais , Poluentes Atmosféricos/toxicidade , LipidômicaRESUMO
Age related cataracts (ARC) represent the main reason for blindness globally. The lens epithelial cells (LECs) participate not only in the metabolism of many substances in the lens but also in maintaining lens transparency. This study used lipidomics to investigate the metabolic differences in LECs of ARC patients with different severity, aiming at identifying potential metabolic biomarkers of ARC. Patients diagnosed with ARC and underwent cataract surgery at Shanghai Tongren Hospital were selected to participate in this study, which were classified as mild ARC group and severe ARC group. During their cataract surgery, anterior lens capsules(LCs) containing LECs were obtained. The lipidomics of LECs were analyzed using the liquid chromatographymass spectrometry (LC-MS). Potential pathways of lipids were searched for using databases such as the Kyoto Encyclopedia of Genes and Genomes (KEGG) and MetaboAnalyst platform. In LEC lipids, 26 lipids have been identified as potential biomarkers between mild ARC and severe ARC, with AUC values of 0.67-0.94. The pathway analysis results revealed that the Glycerophospholipid (GPL) metabolism was significantly influenced, indicating that these metabolic markers contribute significantly to regulating this pathway. The LEC metabolic spectrum demonstrates a proficient ability to differentiate between patients with varying levels of cataracts. Herein, we have successfully identified potential metabolic biomarkers and pathways that have proven to be valuable in enhancing our understanding of ARC pathogenesis. The finding has translational value for developing new cataract treatment methods in the future.
Assuntos
Catarata , Células Epiteliais , Cristalino , Lipidômica , Humanos , Catarata/metabolismo , Catarata/patologia , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Masculino , Feminino , Idoso , Cristalino/metabolismo , Cristalino/patologia , Biomarcadores/metabolismo , Biomarcadores/análise , Pessoa de Meia-Idade , Metabolismo dos Lipídeos , Cromatografia LíquidaRESUMO
CD1 isoforms are MHC class I-like molecules that present lipid-antigens to T cells and have been associated with a variety of immune responses. The lipid repertoire bound and presented by the four CD1 isoforms may be influenced by factors such as the cellular lipidome, subcellular microenvironment, and the properties of the binding pocket. In this study, by shotgun mass spectrometry, we performed a comprehensive lipidomic analysis of soluble CD1 molecules. We identified 1040 lipids, of which 293 were present in all isoforms. Comparative analysis revealed that the isoforms bind almost any cellular lipid.CD1a and CD1c closely mirrored the cellular lipidome, while CD1b and CD1d showed a preference for sphingolipids. Each CD1 isoform was found to have unique lipid species, suggesting some distinct roles in lipid presentation and immune responses. These findings contribute to our understanding of the role of CD1 system in immunity and could have implications for the development of lipid-based therapeutics.
Assuntos
Antígenos CD1 , Lipidômica , Antígenos CD1/metabolismo , Antígenos CD1/imunologia , Humanos , Apresentação de Antígeno/imunologia , Lipídeos/imunologia , Metabolismo dos Lipídeos , Isoformas de Proteínas/imunologia , Antígenos CD1d/metabolismo , Antígenos CD1d/imunologiaRESUMO
A significant hurdle in untargeted lipid/metabolomics research lies in the absence of reliable, cross-validated spectral libraries, leading to a considerable portion of LC-MS features being labeled as unknowns. Despite continuous advancement in annotation tools and libraries, it is important to safeguard, publish and share acquired data through public repositories. Embracing this trend of data sharing not only promotes efficient resource utilization but also paves the way for future repurposing and in-depth analysis; ultimately advancing our comprehension of Covid-19 and other diseases. In this work, we generated an extensive MS-dataset of 39 Covid-19 infected patients versus age- and gender-matched 39 healthy controls. We implemented state of the art acquisition techniques including IDA and SWATH-DIA to ensure a thorough insight in the lipidome and metabolome, ensuring a repurposable dataset.
Assuntos
COVID-19 , Lipidômica , Metabolômica , SARS-CoV-2 , Humanos , COVID-19/metabolismo , Estudos de Casos e Controles , Metaboloma , Espectrometria de Massas , Feminino , Masculino , Cromatografia LíquidaRESUMO
Obligate intracellular protozoan parasite, Leishmania donovani, causative agent of visceral leishmaniasis, led to impaired macrophage functions. It is well documented that many of these changes were induced by parasite-mediated reduction in macrophage cholesterol content. Leishmania-mediated alteration in the other lipids has not been explored in detail yet. Here, we found that the expression of key cholesterol biosynthetic genes and total cellular cholesterol were reduced during L. donovani infection. Further, we have also identified that this reduction in the cholesterol led to increased membrane fluidity and inhibition of antigen-presenting potential of macrophages. In addition to this, we studied the relative changes in different lipids in THP-1-derived macrophages during L. donovani infection through liquid chromatography-mass spectrometry. We found that Sphingomyelin (16:0) and ceramide (20:1, 26:0 and 26:1) were significantly reduced in infected macrophages. We further observed that the majority of different sub-classes of phospholipids were downregulated significantly. Overall ratio of phosphatidylcholine versus phosphotidylethanolamine was decreased which indicated the compensatory mechanism of cell in response to cholesterol reduction. The observed Leishmania-mediated alteration in macrophage-lipidome provided the novel insights into mechanism of host-pathogen interactions.
Assuntos
Colesterol , Leishmania donovani , Leishmaniose Visceral , Lipidômica , Macrófagos , Leishmania donovani/imunologia , Macrófagos/imunologia , Macrófagos/parasitologia , Macrófagos/metabolismo , Humanos , Leishmaniose Visceral/imunologia , Leishmaniose Visceral/parasitologia , Leishmaniose Visceral/metabolismo , Colesterol/metabolismo , Células THP-1 , Interações Hospedeiro-Patógeno/imunologia , Metabolismo dos Lipídeos , Fluidez de MembranaRESUMO
LipidOne 2.0 (https://lipidone.eu) is a new web bioinformatic tool for the analysis of lipidomic data. It facilitates the exploration of the three structural levels of lipids: classes, molecular species, and lipid building blocks (acyl, alkyl, or alkenes chains). The tool's flexibility empowers users to seamlessly include or exclude experimental groups and lipid classes at any stage of the analysis. LipidOne 2.0 offers a range of mono- and multivariate statistical analyses, specifically tailored to each structural level. This includes a novel lipid biomarker identification function, integrating four diverse statistical parameters. LipidOne 2.0 incorporates Lipid Pathway analysis across all three structural levels of lipids. Users can identify lipid-involved reactions through case-control comparisons, generating lists of genes/enzymes and their activation states based on Z scores. Accessible without the need for registration, LipidOne 2.0 provides a user-friendly and efficient platform for exploring and analyzing lipidomic data. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Dataset preparation for LipidOne 2.0 Support Protocol: Lipid nomenclature from spectrometric experiments Basic Protocol 2: Uploading a dataset into LipidOne 2.0 Basic Protocol 3: Data mining of lipidomic dataset by LipidOne 2.0.
Assuntos
Lipidômica , Lipídeos , Lipidômica/métodos , Lipídeos/química , Lipídeos/análise , Internet , Software , Biologia Computacional/métodos , Humanos , Mineração de Dados/métodos , Biomarcadores/análiseRESUMO
To elucidate the lipidomic and metabolomic alterations associated with hypertrophic cardiomyopathy (HCM) pathogenesis, we utilized cmybpc3-/- zebrafish model. Fatty acid profiling revealed variability of 10 fatty acids profiles, with heterozygous (HT) and homozygous (HM) groups exhibiting distinct patterns. Hierarchical cluster analysis and multivariate analyses demonstrated a clear separation of HM from HT and control (CO) groups related to cardiac remodeling. Lipidomic profiling identified 257 annotated lipids, with two significantly dysregulated between CO and HT, and 59 between HM and CO. Acylcarnitines and phosphatidylcholines were identified as key contributors to group differentiation, suggesting a shift in energy source. Untargeted metabolomics revealed 110 and 53 significantly dysregulated metabolites. Pathway enrichment analysis highlighted perturbations in multiple metabolic pathways in the HM group, including nicotinate, nicotinamide, purine, glyoxylate, dicarboxylate, glycerophospholipid, pyrimidine, and amino acid metabolism. Our study provides comprehensive insights into the lipidomic and metabolomic unique signatures associated with cmybpc3-/- induced HCM in zebrafish. The identified biomarkers and dysregulated pathways shed light on the metabolic perturbations underlying HCM pathology, offering potential targets for further investigation and potential new therapeutic interventions.