Untargeted Urinary Metabolomics and Children's Exposure to Secondhand Smoke: The Influence of Individual Differences

Abstract

Children's exposure to secondhand smoke (SHS) is a severe public health problem. There is still a lack of evidence regarding panoramic changes in children's urinary metabolites induced by their involuntary exposure to SHS, and few studies have considered individual differences. This study aims to clarify the SHS-induced changes in urinary metabolites in preschool children by using cross-sectional and longitudinal metabolomics analyses. Urinary metabolites were quantified by using untargeted ultra high-performance liquid chromatography-mass spectrometry (UPLC^(c)-MS/MS). Urine cotinine-measured SHS exposure was examined to determine the exposure level. A cross-sectional study including 17 children in a low-exposure group, 17 in a medium-exposure group, and 17 in a high-exposure group was first conducted. Then, a before-after study in the cohort of children was carried out before and two months after smoking-cessation intervention for family smokers. A total of 43 metabolites were discovered to be related to SHS exposure in children in the cross-sectional analysis (false discovery rate (FDR) corrected p < 0.05, variable importance in the projection (VIP) > 1.0). Only three metabolites were confirmed to be positively associated with children's exposure to SHS (FDR corrected p < 0.05) in a follow-up longitudinal analysis, including kynurenine, tyrosyl-tryptophan, and 1-(3-pyridinyl)-1,4-butanediol, the latter of which belongs to carbonyl compounds, peptides, and pyridines. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis indicated that 1-(3-pyridinyl)-1,4-butanediol and kynurenine were significantly enriched in xenobiotic metabolism by cytochrome P450 (p = 0.040) and tryptophan metabolism (p = 0.030), respectively. These findings provide new insights into the pathophysiological mechanism of SHS and indicate the influence of individual differences in SHS-induced changes in urinary metabolites in children.

Keywords: children; cotinine; metabolomics; secondhand smoke; untargeted.

Figure 1

Flowchart of included children at the baseline and follow-up. LOD, limit of detection.

Figure 2

Orthogonal partial least-squares discriminant analysis (OPLS-DA) score plots for BH and BL groups, as well as for BM and BL groups, in positive-ion and negative-ion modes. Abbreviations: BH, high-level SHS exposure at the baseline; BM, medium-level SHS exposure at the baseline; and BL, low-level SHS exposure at the baseline.

Figure 3

Heatmap of the levels of differentially expressed metabolites in each group at the baseline.

Figure 4

Percent change of the three differential metabolites at post-intervention compared to pre-intervention for each child from the ID group. Blue, tyrosyl-tryptophan; red, 1-(3-pyridinyl)-1,4-butanediol; green, kynurenine; and ID, concentration of nicotine in children’s urine was decreased after the smoking-cessation intervention for smokers, which is called the intervention-declined group (ID) for short.

Figure 5

Simplified schematic of the relationship between exposure to SHS and metabolites following the KEGG pathway database.