MetaboLights

application/xml

ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12848/m_MTBLS12848_NMR___metabolite_profiling_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12848/s_MTBLS12848.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12848/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12848/a_MTBLS12848_NMR___metabolite_profiling.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS12848Topspin version 3.0 (Bruker Biospin) was used for spectrometer control and data pro-cessing. Spectra analysis employed an untargeted metabolomic approach, where each metabolite was identified prior to statistical testing using Chenomx NMR-Suite v8.0 (Chenomx NMR Suite, v8.0, Edmonton, AB, Canada) (https://www.chenomx.com/). Quantitative analysis of the 1D-NMR spectra was performed using NMRProcFlow [33], and the resulting data matrix was subsequently analyzed using statistical tools.MetaboLightsPublicNuclear Magnetic Resonance (NMR) -Effects of nicotine on SH-SY5Y cells: an NMR-based metabolomic study.1D 1H NMR spectra were recorded on a Bruker Ascend™ 600 MHz spectrometer equipped with a 5 mm triple resonance Z gradient TXI probe (Bruker Co, Rheinstetten, Germany) at 298 K. 1D NOESY NMR spectra were recorded with 20 k points, 12 ppm spectral width, 1.36s acquisition time, 5 s relaxation delay, 10 ms of mixing time and 128 scans [32]. Topspin version 3.0 (Bruker Biospin) was used for spectrometer control and data processing. University of SalernoCarmen MarinoexometabolomeendometabolomeNMR spectroscopy assayTo extract intracellular metabolites from cell pellet, homogenization was followed by bi-phasic extraction method using methanol, chloroform, and water in a 1:1:1 ratio [31]. Samples were centrifuged at 6000 rpm for 10 minutes at 4 °C to separate the polar and apolar phases. Polar extracts from cell pellet were dried under vacuum with a SP-Genevac EZ-2 4.0 concentrator, while lipophilic extracts were dried with nitrogen flow for later analysis. All extracts were stored at −80 °C before NMR testing.Homo sapienshttps://www.ebi.ac.uk/metabolights/MTBLS12848Carmen Marino. University of Salerno. cmarino@unisa.it.Anna Maria D'Ursi. University of Salerno. dursi@unisa.it.Enza Napolitano. University of Salerno. enapolitano@unisa.it.Sample data were normalized using the sum, then log-transformed and Pareto scaled, and analysed with the MixOmics R package (mixOmics package) and Metaboanalyst 6.0 [33,34]. Univariate analysis was performed separately on the exometabolome and endometabolome of the groups, using T-test and Fold Change, and results were displayed in a Volcano plot [35]. To enhance the accuracy of the data and gain biological insights, multivariate statistical analysis (MVA) was first applied to the exometabolome and endometabolome concentration matrices, followed by an analysis of the combined data sets. CompartmentType of treatmentcmarino@unisa.itTo prepare metabolomic samples, cells were plated in 60 mm culture dishes and al-lowed to adhere overnight. Then, 100 µM nicotine was added for 24 hours. For the control group, cells were treated only with vehicle (water) for the same duration. All experimental conditions were tested using four biological replicates, each comprising three technical replicates.After treatments, both culture media and cell pellet were collected for the metabolom-ics analysis targeting the exometabolome and endometabolome, respectively. Specifically, the medium was transferred to microcentrifuge tubes and centrifuged at 1000× g for 10 minutes. The same procedure was applied to cell-free medium incubated under identical conditions. The resulting supernatants were transferred to fresh microcentrifuge tubes and stored at -80°C until NMR analysis. After media removal, cell dishes were washed with cold PBS (pH 7.4) to remove media residues and cells were collected by scraping in meth-anol To extract intracellular metabolites from cell pellet, homogenization was followed by biphasic extraction method using methanol, chloroform, and water in a 1:1:1 ratio [31]. Samples were centrifuged at 6000 rpm for 10 minutes at 4 °C to separate the polar and apolar phases. Polar extracts from cell pellet were dried under vacuum with a SP-Genevac EZ-2 4.0 concentrator, while lipophilic extracts were dried with nitrogen flow for later analysis. All extracts were stored at −80 °C before NMR testing.1D 1H NMR spectra were recorded on a Bruker Ascend™ 600 MHz spectrometer equipped with a 5 mm triple resonance Z gradient TXI probe (Bruker Co, Rheinstetten, Germany) at 298 K. 1D NOESY NMR spectra were recorded with 20 k points, 12 ppm spectral width, 1.36s acquisition time, 5 s relaxation delay, 10 ms of mixing time and 128 scans [32]. Topspin version 3.0 (Bruker Biospin) was used for spectrometer control and data processing.MetabolomicsMetabolomicsnuclear magnetic resonance spectroscopyuntargeted metabolitesNicotineMetabolomicsnuclear magnetic resonance spectroscopyuntargeted metabolitesNicotineLyophilized cell extracts were reconstituted in 200 μL of buffer (50 mM Na2HPO4, 1 mM trimethylsilyl propionic-2,2,3,3-d4 acid sodium salt (TSP-d4), 10% of D2O). TSP-d4 was used as an internal standard for the alignment and quantification of NMR signals. For growth media analysis, 100 μL of cell medium was mixed with 100 μL of the same buffer used for the lyophilized extracts. The resulting samples were transferred into 3 mm NMR tubes for 1H NMR acquisition.TaurineLactatesarcosineAcetyl cysteine2-methyl-3-ketovaleric acidarginine3-methyl-2-oxovalerateglycineglutamateTyrosineThreonineSuccinate3-hydroxybutyrate5,6-dihydrothymineAcetatelysinehomocystinemethionine2-aminoisobutyric acidUDP-glucosedimethylallyl pyrophosphateSerineglutathioneProlineLeucineglutaminemethylmalonatecholineformateIsovalerateisobutyryl-L-carnitine2-oxobutyrateciticolinephosphoryl cholinemethanolriboflavinIsoleucinehomocysteineAlanineglucoseUDP N-acetylglucosaminePhenylalanineN-acetyl-L-aspartateTryptophanAspartatepyroglutamatepyruvateFructose2-hydroxybutyrateglycerophosphocholineLactoseacetoacetateValinecarnitinebetaineATPhistidinefalseEffects of nicotine on SH-SY5Y cells: an NMR-based metabolomic studyNicotine is a naturally occurring alkaloid primarily found in Nicotiana tabacum. This phytochemical is well known for its addictive properties, and its consumption—particularly through tobacco smoking—is strongly associated with an increased risk of malignancies, metabolic dysfunctions, and cardiovascular as well as respiratory diseases. Despite these adverse effects, several studies have also reported beneficial actions of nicotine, including the enhancement of cognitive functions in several neurodegenerative diseases. To better elucidate the multiple effects of nicotine and clarify their underlying mechanisms, we performed an NMR-based metabolomic analysis of SH-SY5Y neuroblastoma cells exposed to nicotine action. Our results indicate that nicotine modulates mitochondrial function and membrane turnover, thereby influencing mitochondrial bioenergetics, synaptic plasticity, and connectivity. Collectively, these findings may contribute, at least in part, to explain the neuroprotective effects of nicotine described in preclinical models of neurodegenerative disease.2026-06-262025-08-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:86365CHEBI:1148CHEBI:30831CHEBI:20067CHEBI:27468CHEBI:15366CHEBI:15344CHEBI:28939CHEBI:16449CHEBI:29016CHEBI:35391CHEBI:15422CHEBI:17750CHEBI:17126CHEBI:15354CHEBI:16436CHEBI:30751CHEBI:28757CHEBI:17234CHEBI:14321CHEBI:28300CHEBI:16856CHEBI:36313CHEBI:15428CHEBI:27570CHEBI:17230CHEBI:17485CHEBI:84838CHEBI:24898CHEBI:28484CHEBI:78320CHEBI:17716CHEBI:25017CHEBI:25094CHEBI:17790CHEBI:16811CHEBI:30861CHEBI:21547CHEBI:28044CHEBI:26271CHEBI:16010CHEBI:32816CHEBI:17015CHEBI:15611CHEBI:17822CHEBI:26806CHEBI:15891CHEBI:26986CHEBI:27897CHEBI:18186CHEBI:18066CHEBI:27266CHEBI:35932CHEBI:27971CHEBI:16057CHEBI:18132CHEBI:16264