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Article Open Access

Metabolomic profiling reveals amino acid dysregulation in congenital heart disease: Arginine‑induced embryonic malformations and pathogenic insights

  • Authors:
    • Zhen Li
    • Can Cui
    • Jinping Li
    • Guangrui Lai
    • Chong Qiao
  • View Affiliations / Copyright

    Affiliations: Department of Obstetrics and Gynecology, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004, P.R. China, Department of Clinical Genetics, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004, P.R. China
    Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 159
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    Published online on: July 25, 2025
       https://doi.org/10.3892/br.2025.2037
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Abstract

Congenital heart disease (CHD) is the most common form of malformation seen in China. In most cases, its pathogenesis is unclear. The present study aimed to identify the differential metabolites and novel screening markers to discover the potential pathogenesis of CHD. Cultured amniotic fluid cells from pregnant patients carrying CHD‑affected (n=24) or healthy (n=24) fetuses were collected. Untargeted metabolomics was performed on the cells. A total of 292 metabolites (172 up‑ and 120 downregulated) were significantly different between the CHD and control groups (variable importance on projection >1 and P<0.05). Significantly different metabolites were screened and analyzed using hierarchical clustering and Kyoto Encyclopedia of Genes and Genomes metabolic pathway enrichment. These data demonstrated that amino acid metabolism was considerable elevated. The most notable differential metabolites were lysine and arginine, suggesting they may play an important role in heart development. When arginine were used to treat pregnant mice, embryos (embryonic day 15.5) demonstrated increased malformation rate, the litter size decreased (4.3±1.03 vs. 6.5±1.05, P<0.01) and the naked deformity rate significantly increased (30.8 vs. 0.0%, P<0.001), compared with the controls. The embryonal heart cavity was larger and the heart wall became thinner. The present study suggested that amino acid metabolites may serve a crucial role in heart development and could serve as potential screening markers for CHD. Additionally, the adverse effects of arginine treatment on embryonic development highlight its role in CHD pathogenesis.
View Figures

Figure 1

Experimental workflow. Amniotic fluid
cells were obtained by centrifugation and cultured for 1 week for
karyotype analysis. The culture medium transferred and cultured for
1 week. Cells cultured for 2 weeks that would otherwise be
discarded were used for metabolomic analysis.

Figure 2

Representative metabolic profile and
experimental quality evaluation. (A) Representative TIC from sample
in electrospray ionization positive mode. (B) TIC overlapping map
of positive modes of quality control sample. (C) Principal
component analysis score chart of samples. TIC, total ion
chromatogram; QC, quality control; CHD, congenital heart disease;
RT, retention time.

Figure 3

Significant differential metabolite
analysis. (A) OPLS-DA between control and CHD groups. The values of
R2Y and Q2 demonstrated the goodness of fit and predictability of
the model, respectively. (B) Permutation test of the OPLS-DA model.
The intercept of Q2 suggested no overfitting in the model. OPLS-DA,
orthogonal partial least squares discriminant analysis; CHD,
congenital heart diseas.

Figure 4

Volcano plot of CHD vs. control.
Association between fold-change of CHD vs. control and significance
of the metabolic features. Upregulated, fold-change >2 and
P<0.05; downregulated, fold-change <0.5 and P<0.05. CHD,
congenital heart diseas.

Figure 5

KEGG pathway enrichment analysis of
the differential metabolites. Amino acids (L-lysine, -arginine,
-glutamine, -ornithine and -asparagine) were involved in ‘ABC
transporters’, ‘biosynthesis of amino acids’, ‘purine metabolism’,
‘protein digestion and absorption’, ‘D-arginine and D-ornithine
metabolism’ and ‘amoebiasis’. KEGG, Kyoto Encyclopedia of Genes and
Genomes; ABC, ATP-binding cassette.

Figure 6

Arg results in heart malformation but
Lys does not induce notable abnormality. (A) Abnormal embryos were
found in Arg-treated group. (B) Maternal plasma Arg concentration
following Arg treatment. (C) HE staining suggested embryos had
enlarged heart cavities and thinner heart walls following Arg
treatment. HE, hematoxylin-eosin.
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Copy and paste a formatted citation
Spandidos Publications style
Li Z, Cui C, Li J, Lai G and Qiao C: Metabolomic profiling reveals amino acid dysregulation in congenital heart disease: Arginine‑induced embryonic malformations and pathogenic insights. Biomed Rep 23: 159, 2025.
APA
Li, Z., Cui, C., Li, J., Lai, G., & Qiao, C. (2025). Metabolomic profiling reveals amino acid dysregulation in congenital heart disease: Arginine‑induced embryonic malformations and pathogenic insights. Biomedical Reports, 23, 159. https://doi.org/10.3892/br.2025.2037
MLA
Li, Z., Cui, C., Li, J., Lai, G., Qiao, C."Metabolomic profiling reveals amino acid dysregulation in congenital heart disease: Arginine‑induced embryonic malformations and pathogenic insights". Biomedical Reports 23.4 (2025): 159.
Chicago
Li, Z., Cui, C., Li, J., Lai, G., Qiao, C."Metabolomic profiling reveals amino acid dysregulation in congenital heart disease: Arginine‑induced embryonic malformations and pathogenic insights". Biomedical Reports 23, no. 4 (2025): 159. https://doi.org/10.3892/br.2025.2037
Copy and paste a formatted citation
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Spandidos Publications style
Li Z, Cui C, Li J, Lai G and Qiao C: Metabolomic profiling reveals amino acid dysregulation in congenital heart disease: Arginine‑induced embryonic malformations and pathogenic insights. Biomed Rep 23: 159, 2025.
APA
Li, Z., Cui, C., Li, J., Lai, G., & Qiao, C. (2025). Metabolomic profiling reveals amino acid dysregulation in congenital heart disease: Arginine‑induced embryonic malformations and pathogenic insights. Biomedical Reports, 23, 159. https://doi.org/10.3892/br.2025.2037
MLA
Li, Z., Cui, C., Li, J., Lai, G., Qiao, C."Metabolomic profiling reveals amino acid dysregulation in congenital heart disease: Arginine‑induced embryonic malformations and pathogenic insights". Biomedical Reports 23.4 (2025): 159.
Chicago
Li, Z., Cui, C., Li, J., Lai, G., Qiao, C."Metabolomic profiling reveals amino acid dysregulation in congenital heart disease: Arginine‑induced embryonic malformations and pathogenic insights". Biomedical Reports 23, no. 4 (2025): 159. https://doi.org/10.3892/br.2025.2037
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