Protective effect of L-pipecolic acid on constipation in C57BL/6 mice based on gut microbiome and serum metabolomic

Study information

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study design

laboratory experiment

Citation

PMID PubMed identifier for scientific articles.

37210496

DOI Digital object identifier for electronic documents.

10.1186/s12866-023-02880-3.

URI

https://bmcmicrobiol.biomedcentral.com/articles/10.1186/s12866-023-02880-3

Authors

Li H, Xiao HY, Yuan LP, Yan B, Pan Y, Tian PP, Zhang WJ

Journal

BMC microbiology

Year

2023

Keywords:

Children, Functional constipation, Gut microbiota, L-pipecolic acid, Metabolite profiles

BACKGROUND: Functional constipation (FC) in children affects their growth, development and quality of life. L-pipecolic acid (L-PA) was decreased in FC children based on gut microbiome and serum metabolomic. In this study, loperamide-induced constipation in mice was used to evaluate the effects of L-PA on constipated mice. METHOD: 26 FC and 28 healthy children were recruited. Stool samples and serum samples were subjected to 16S rDNA sequencing and ultra-performance liquid chromatography/quadrupole time of flight (UPLC-Q/TOF-MS) approach, respectively. A loperamide-induced mouse constipation model was developed, and all mice were randomly divided into control (Con), loperamide (Lop) and L-PA (Lop + L-PA) treatment groups (6 mice per group). The mice in the Lop + L-PA group were given L-PA (250 mg/kg, once a day) and loperamide; the Lop group was given loperamide for 1 week, and the Con group was given saline. The fecal parameters and intestinal motility of mice in each group were detected. serum 5-HT levels and colon 5-HT expression were detected by ELISA and immunohistochemistry, respectively; qRT-PCR was used to detect the expression of AQP3 and 5-HT4R mRNA in each group. RESULTS: 45 differential metabolites and 18 significantly different microbiota were found in FC children. The α and β diversity of gut microbiota in FC children was significantly reduced. Importantly, serum L-PA was significantly reduced in FC children. The KEGG pathway enrichment were mainly enriched in fatty acid biosynthesis, lysine degradation, and choline metabolism. L-PA was negatively associated with Ochrobactrum, and N6, N6, N6-trimethyl-l-lysine was positively associated with Phascolarcrobacterium. In addition, L-PA improved the fecal water content, intestinal transit rate, and increased the serum 5-HT levels in constipated mice. Moreover, L-PA increased the expression of 5-HT4R, reduced AQP3, and regulated constipation-associated genes. CONCLUSIONS: Gut microbiota and serum metabolites were significantly altered in children with FC. The abundance of Phascolarctobacterium and Ochrobactrum and serum L-PA content were decreased in FC children. L-PA was found to alleviate the fecal water content, increase intestinal transit rate and the first black stool defecation time. L-PA improved constipation by increasing 5-HT and 5-HT4R expression while down-regulating AQP3 expression.

Experiment 1

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Curated date: 2025/03/14

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Subjects

Location of subjects: China

Host species Species from which microbiome was sampled. Contact us to have more species added.: Mus musculus

Body site Anatomical site where microbial samples were extracted from according to the Uber Anatomy Ontology: Colon Hindgut,Large bowel,Posterior intestine,Colon,colon

Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology: Constipation Constipation,Costiveness,Dyschezia,constipation

Group 0 name Corresponds to the control (unexposed) group for case-control studies: Healthy Children

Group 1 name Corresponds to the case (exposed) group for case-control studies: Functional Constipation(FC)

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group: FC is a common gastrointestinal disease among children and a major health problem around the globe. In recent years, gut microbiota and metabolites of microbiome have been reported to be involved in FC mechanisms. Kim et al. determined that the microbiota of FC patients had a decreased abundance of Prevotella and increased abundance of Coprococcus, Ruminococcus, Blautia, Anaerotruncus, and Clostridium.

Group 0 sample size Number of subjects in the control (unexposed) group: 28

Group 1 sample size Number of subjects in the case (exposed) group: 26

Antibiotics exclusion Number of days without antibiotics usage (if applicable) and other antibiotics-related criteria used to exclude participants (if any): 3 months.

Lab analysis

Sequencing type: 16S

16S variable region One or more hypervariable region(s) of the bacterial 16S gene: V3-V4

Statistical Analysis

Data transformation Data transformation applied to microbial abundance measurements prior to differential abundance testing (if any).: relative abundances

Statistical test: T-Test

Significance threshold p-value or FDR threshold used for differential abundance testing (if any): 0.05

Alpha Diversity

Chao1 Abundance-based estimator of species richness: decreased

Signature 1

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Source: Figure1F

Description: Differential analysis between functional constipation and Healthy Children using STAMP Analysis

Abundance in Group 1: increased abundance in Functional Constipation(FC)

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Klebsiella

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Source: Figure 1F

Description:

Abundance in Group 1: decreased abundance in Functional Constipation(FC)

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Phascolarctobacterium

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Experiment 2

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Differences from previous experiment shown

Subjects

Body site Anatomical site where microbial samples were extracted from according to the Uber Anatomy Ontology: Colon , Blood serum Hindgut,Large bowel,Posterior intestine,Colon,colon,Serum,Blood serum,blood serum

Group 1 name Corresponds to the case (exposed) group for case-control studies: Functional constipation

Antibiotics exclusion Number of days without antibiotics usage (if applicable) and other antibiotics-related criteria used to exclude participants (if any): 3 Months

Lab analysis

16S variable region One or more hypervariable region(s) of the bacterial 16S gene: Not specified

Statistical Analysis

Statistical test: Not specified