Summary of Study ST002998

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001868. The data can be accessed directly via it's Project DOI: 10.21228/M8842H This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Perform statistical analysis  |  Show all samples  |  Show named metabolites  |  Download named metabolite data  
Download mwTab file (text)   |  Download mwTab file(JSON)   |  Download data files (Contains raw data)
Study IDST002998
Study TitleThe role of gut microbiota in muscle mitochondria function, colon health, and sarcopenia: from clinical to bench
Study SummaryTo further investigate if metabolites produced by probiotics could directly improve muscle growth, an in-vitro study was performed by utilizing C2C12 myoblasts and myotubes, as well as probiotic supernatant. The supernatant of probiotics was collected and LC-MS analysis was performed.
Institute
The Chinese University of Hong Kong
Last NameWong
First NamePui Yan
Address30-32 Ngan Shing Street , Shatin, New Territories
Emailpuiyanwong@cuhk.edu.hk
Phone(852)-35052756
Submit Date2023-11-29
Raw Data AvailableYes
Raw Data File Type(s)mzXML
Analysis Type DetailGC/LC-MS
Release Date2024-12-30
Release Version1
Pui Yan Wong Pui Yan Wong
https://dx.doi.org/10.21228/M8842H
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:


Project:

Project ID:PR001868
Project DOI:doi: 10.21228/M8842H
Project Title:From clinical to benchside: Lacticaseibacillus and Faecalibacterium are positively associated with muscle health and alleviate age-related muscle disorder
Project Summary:Sarcopenia is an age-related muscle disorder that increases the risk of adverse clinical outcomes, but its pathogenesis is unclear and treatments are limited. Increasing evidence shows that gut microbiota is potentially associated with sarcopenia. To investigate its role in sarcopenia, we (i) compared gut microbiota and metabolite composition between older persons with and without sarcopenia, (ii) performed fecal microbiota transplantation (FMT) from human donors to antibiotic-treated mice, and (iii) identified two specific probiotics for treatment of sarcopenia in aged mice. Metagenomic and metabolomic analyses showed that people with sarcopenia had different microbial composition and metabolites, and that fecal purine could accurately identify sarcopenia. After FMT, mice receiving microbes from people with sarcopenia displayed lower muscle mass and strength. Correlation analysis revealed Lacticaseibacillus rhamnosus (LR) and Faecalibacterium prausnitzii (FP) were positively related to muscle health in old people. Both probiotic LR, FP and their combination enhanced muscle mass, function, and fiber type proportion of aged mice. Transcriptomics showed that genes related to tricarboxylic acid cycle were enriched after treatment. Mitochondria density, muscle ATP content, NAD+/NADH, proteins related to mitochondrial dynamics and biogenesis were improved by both probiotics. In in-vitro studies, probiotic-conditional medium (PCM) containing FP supernatant or the combination of FP and LR supernatants enhanced proliferation of C2C12 myoblasts, whilst LR PCM alone did not. The mechanisms of LR may be related to colon health improvement. Results showed gut microbiota dysbiosis is one of pathogenic factors of sarcopenia, and muscle-related probiotics could alleviate age-related muscle disorders. Further clinical translation is warranted.
Institute:The Chinese University of Hong Kong
Department:ORT
Laboratory:The Prince of Wales Hospital
Last Name:Wong
First Name:Pui Yan
Address:30-32 Ngan Shing Street , Shatin, New Territories
Email:puiyanwong@cuhk.edu.hk
Phone:(852)-35052756

Subject:

Subject ID:SU003111
Subject Type:Bacteria
Subject Species:Lacticaseibacillus rhamnosus and Faecalibacterium prausnitzii
Species Group:Bacteria

Factors:

Subject type: Bacteria; Subject species: Lacticaseibacillus rhamnosus and Faecalibacterium prausnitzii (Factor headings shown in green)

mb_sample_id local_sample_id Treatment
SA326432FP3FP
SA326433FP2FP
SA326434FP1FP
SA326435FP_medium_3FP_medium
SA326436FP_medium_1FP_medium
SA326437FP_medium_2FP_medium
SA326438LGG2LGG
SA326439LGG3LGG
SA326440LGG1LGG
SA326441LGG_medium_3LGG_medium
SA326442LGG_medium_1LGG_medium
SA326443LGG_medium_2LGG_medium
Showing results 1 to 12 of 12

Collection:

Collection ID:CO003104
Collection Summary:The Lacticaseibacillus rhamnosus ATCC 53103 was cultured on MRS Agar (Sigma, USA), and a single colony of LR was cultured in DeMan-Rogosa-Sharpe (MRS, Sigma) broth to obtain 108 colony forming unit (CFU)/mL in the anaerobic chamber (Bugbox Plus UM-017, Baker). After centrifugating at 1600g for 10 min at 4 ℃ and discarding supernatant, the pellet was resuspended in sterile water, centrifuged again, and the supernatant was discarded. The pellet was resuspended in 1 mL to obtain 1010 CFU/mL. The CFU was enumerated by culturing 10 μL of the aliquot by serial dilution method in Blood agar. The Faecalibacterium prausnitzii ATCC27768 was cultured on freshly prepared pre-reduced YBHA media. After 48h incubation at 37 ℃ in the anaerobic chamber, a single colony from the YBHA plate was cultured in the YBHI media to obtain 108/mL. The dilution and enumeration procedures were as above.
Sample Type:Bacterial cells

Treatment:

Treatment ID:TR003120
Treatment Summary:The medium and supernatant of probiotics (LR and FP) were obtained from three separate preparations (n = 3 per group). 100 μL of sample was transferred to an EP tube. After the addition of 400 μL of extract solution (methanol: acetonitrile= 1: 1, containing isotopically-labelled internal standard mixture), the samples were vortexed for 30 s, sonicated for 10 min in ice-water bath, and incubated for 1 h at -40 ℃ to precipitate proteins. Then the sample was centrifuged at 12000 rpm(RCF=13800(×g),R= 8.6cm) for 15 min at 4 ℃. The resulting supernatant was transferred to a fresh glass vial for analysis. The quality control (QC) sample was prepared by mixing an equal aliquot of the supernatants from all of the samples.

Sample Preparation:

Sampleprep ID:SP003117
Sampleprep Summary:The medium and supernatant of probiotics (LR and FP) were obtained from three separate preparations (n = 3 per group). 100 μL of sample was transferred to an EP tube. After the addition of 400 μL of extract solution (methanol: acetonitrile= 1: 1, containing isotopically-labelled internal standard mixture), the samples were vortexed for 30 s, sonicated for 10 min in ice-water bath, and incubated for 1 h at -40 ℃ to precipitate proteins. Then the sample was centrifuged at 12000 rpm(RCF=13800(×g),R= 8.6cm) for 15 min at 4 ℃. The resulting supernatant was transferred to a fresh glass vial for analysis. The quality control (QC) sample was prepared by mixing an equal aliquot of the supernatants from all of the samples.

Combined analysis:

Analysis ID AN004924 AN004925
Analysis type MS MS
Chromatography type Unspecified Unspecified
Chromatography system Thermo Vanquish Thermo Vanquish
Column Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um) Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um)
MS Type ESI ESI
MS instrument type QTOF Single quadrupole
MS instrument name Agilent 6545 QTOF 1290 Infinity LC UPLC
Ion Mode NEGATIVE POSITIVE
Units nmol/g nmol/g

Chromatography:

Chromatography ID:CH003717
Instrument Name:Thermo Vanquish
Column Name:Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um)
Column Temperature:4 ℃
Flow Gradient:0-0.5 min, 95% B; 0.5-7 min, 95%-65% B; 7-8 min, 65%-40% B; 8-9 min, 40% B; 9-9.1 min, 40%-95% B; 9.1-12 min, 95% B
Flow Rate:sheath gas flow rate as 50 Arb, Aux gas flow rate as 15 Arb
Solvent A:25 mmol/L ammonium acetate and 25 mmol/L ammonia hydroxide in water(pH = 9.75)
Solvent B:100% acetonitrile
Chromatography Type:Unspecified

MS:

MS ID:MS004667
Analysis ID:AN004924
Instrument Name:Agilent 6545 QTOF
Instrument Type:QTOF
MS Type:ESI
MS Comments:LC-MS/MS analyses were performed using an UHPLC system (Vanquish, Thermo Fisher Scientific) with a UPLC BEH Amide column (2.1 mm × 100 mm, 1.7 μm) coupled to Orbitrap Exploris 120 mass spectrometer (Orbitrap MS, Thermo). The mobile phase consisted of 25 mmol/L ammonium acetate and 25 ammonia hydroxide in water pH = 9.75))(A) and acetonitrile (B). The auto-sampler temperature was 4 ℃℃, and the injection volume was 2 μμL. The Orbitrap Exploris 120 mass spectrometer was used for its ability to acquire MS/MS spectra on information-dependent acquisition (IDA) mode in the control of the acquisition software (Xcalibur, Thermo). In this mode, the acquisition software continuously evaluates the full scan MS spectrum. The ESI source conditions were set as following: sheath gas flow rate as 50 Arb, Aux gas flow rate as 15 Arb, capillary temperature 320 ℃℃, full MS resolution as 60000, MS/MS resolution as 15000 collision energy as 10/30/60 in NCE mode, spray Voltage as 3.8 kV (positive) or -3.4 kV (negative), respectively.
Ion Mode:NEGATIVE
  
MS ID:MS004668
Analysis ID:AN004925
Instrument Name:1290 Infinity LC UPLC
Instrument Type:Single quadrupole
MS Type:ESI
MS Comments:LC-MS/MS analyses were performed using an UHPLC system (Vanquish, Thermo Fisher Scientific) with a UPLC BEH Amide column (2.1 mm × 100 mm, 1.7 μm) coupled to Orbitrap Exploris 120 mass spectrometer (Orbitrap MS, Thermo). The mobile phase consisted of 25 mmol/L ammonium acetate and 25 ammonia hydroxide in water pH = 9.75))(A) and acetonitrile (B). The auto-sampler temperature was 4 ℃℃, and the injection volume was 2 μμL. The Orbitrap Exploris 120 mass spectrometer was used for its ability to acquire MS/MS spectra on information-dependent acquisition (IDA) mode in the control of the acquisition software (Xcalibur, Thermo). In this mode, the acquisition software continuously evaluates the full scan MS spectrum. The ESI source conditions were set as following: sheath gas flow rate as 50 Arb, Aux gas flow rate as 15 Arb, capillary temperature 320 ℃℃, full MS resolution as 60000, MS/MS resolution as 15000 collision energy as 10/30/60 in NCE mode, spray Voltage as 3.8 kV (positive) or -3.4 kV (negative), respectively.
Ion Mode:POSITIVE
  logo