Mouse fecal metabolomics raw data from CTRL, PLN, AmLN, and pAmLN

Published: 16 September 2024| Version 1 | DOI: 10.17632/498c4cbvnt.1
Contributor:
Linyu Peng

Description

This dataset contains the raw fecal metabolomics data generated from a study investigating the effects of live Akkermansia muciniphila (AKK) and pasteurized Akkermansia muciniphila (pAKK) on a preeclampsia (PE) mouse model induced by N(G)-nitro-L-arginine methyl ester (L-NAME). The metabolomic profiling was conducted to assess the metabolic changes across four experimental groups: CTRL (control group), PLN (PE model group with L-NAME), AmLN (PE model with live AKK treatment), and pAmLN (PE model with pAKK treatment).

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1.Animal Model and Grouping: Pregnant C57BL/6J mice (6–8 weeks old) were randomly assigned to four groups: CTRL, PLN, AmLN, and pAmLN. To induce a preeclampsia (PE) model, L-NAME (NG-nitro-L-arginine methyl ester hydrochloride) was administered at 125 mg/kg/day via gavage starting from embryonic day E8.5 for 10 consecutive days. Mice in the CTRL group received only PBS. PLN group mice were treated with L-NAME , while the AmLN and pAmLN groups received L-NAME in combination with either live Akkermansia muciniphila (AKK) or pasteurized AKK (pAKK). 2.Fecal Sample Collection and Preparation: Fecal samples were collected from each mouse group at E18.5, snap-frozen, and stored at -80°C until further analysis. Fecal metabolomics data were generated for all four groups (CTRL, PLN, AmLN, and pAmLN). 3.Metabolite Extraction: Approximately 100 mg of fecal material from each sample was ground in liquid nitrogen. Prechilled 80% methanol was added for extraction, followed by vortexing and centrifugation at 15,000 g for 20 minutes at 4°C. Supernatants were diluted to a final concentration of 53% methanol in LC-MS grade water. These supernatants were subjected to a second centrifugation (15,000 g, 20 minutes, 4°C) before injection into the LC-MS/MS system. 4.UHPLC-MS/MS Analysis: Fecal metabolite profiling was performed using a Vanquish UHPLC system (Thermo Fisher Scientific) coupled with an Orbitrap Q Exactive™ HF mass spectrometer. Samples were injected onto a Hypersil Gold column (100 × 2.1 mm, 1.9 µm), with a 12-minute linear gradient at a flow rate of 0.2 mL/min. Eluents consisted of 0.1% formic acid in water (eluent A) and methanol (eluent B). The mass spectrometer operated in both positive and negative ion modes. A detailed solvent gradient and instrumentation parameters, including ionization conditions, were followed as per the standard operating procedure. 5.Data Processing and Statistical Analysis: Raw LC-MS/MS data were processed using Compound Discoverer 3.3 software to perform peak alignment, detection, and quantification for each metabolite. Principal Component Analysis (PCA) and Partial Least Squares Discriminant Analysis (PLS-DA) were conducted to identify and visualize metabolic differences across the groups. Differential metabolites were identified using univariate analysis (t-test), with a p-value < 0.05 and VIP score > 1. Pathway enrichment analysis was performed using the KEGG database to identify key metabolic pathways affected in the study.

Institutions

Southern Medical University

Categories

Untargeted Metabolomics

Funding

Natural Science Foundation of Guangzhou Municipality

2023A1515011074

Natural Science Foundation of Guangzhou Municipality

2022A1515011730

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