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. 2024 May 30;16(1):2359729. doi: 10.1080/19490976.2024.2359729

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© 2024 The Author(s). Published with license by Taylor & Francis Group, LLC.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent.

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Figure 7. — H₂O₂ ameliorates myelin-related gene expression and D-glucaric acid production in neonatal mice.

(a) H₂O₂ concentration after incubating skim milk with or without amino acids (AA). AA (-) (WT milk n = 6, LAO1 KO milk n = 4, t₈ = 0.83, p = 0.43, Student’s t-test.), AA (+) (WT milk n = 6, LAO1 KO milk n = 4, t₈ = 5.89, p < 0.05, Student’s t-test).

(b) Investigated the impact of orally administrated H₂O₂ to neonatal LAO1 KO mice, and the schematic diagram was illustrated. Created with BioRender.com.

(c) Relative peak intensity of D-glucaric acid from serum metabolomics. n = 8 each, Saline versus H₂O₂ 1 μg/day: q₃₅ = 2.167, p = 0.12, Saline versus H₂O₂ 5 μg/day: q₃₅ = 1.35, p = 0.47, Saline versus H₂O₂ 10 μg/day: q₃₅ = 0.07, p = 0.99, Saline versus H₂O₂ 50 μg/day: q₃₅ = 3.36, p < 0.05, Dunnett’s test.

(d) Quantitative real-time polymerase chain reaction (qRT-PCR) of Mbp and Plp1 using the hippocampus of H₂O₂-supplemented 10-day-old neonates. Mbp (n = 8 each, Saline versus H₂O₂ 1 μg/day: q₃₅ = 1.533, p = 0.36, Saline versus H₂O₂ 5 μg/day: q₃₅ = 0.1739, p = 0.99, Saline versus H₂O₂ 10 μg/day: q₃₅ = 2.21, p = 0.11, Saline versus H₂O₂ 50 μg/day: q₃₅ = 6.006, p < 0.05, Dunnett’s test), Plp1 (n = 8 each, Saline versus H₂O₂ 1 μg/day: q₃₅ = 0.4061, p = 0.98, Saline versus H₂O₂ 5 μg/day: q₃₅ = 1.061, p = 0.67, Saline versus H₂O₂ 10 μg/day: q₃₅ = 0.7489, p = 0.87, Saline versus H₂O₂ 50 μg/day: q₃₅ = 7.767, p < 0.05, Dunnett’s test).

(e and f) Representative myelin basic protein (MBP) staining and quantitative analysis of P10 LAO1 KO mice dorsal hippocampus. Saline n = 5, H₂O₂ 50 μg/day n = 4, t₇ = 3.14, p < 0.05, Student’s t-test.

(g) Beta diversity of the 16S rRNA gene sequencing data sets from Saline and H₂O₂ 50 μg/day-administrated LAO1 KO neonate feces as determined using principal coordinate analysis (PCoA) of Bray-Curtis index. n = 8 each, F = 2.21, R-squared = 0.14, p < 0.05, PERMANOVA.

(h) Alpha-diversity was evaluated by chao1 and Shannon index. chao1 (n = 8 each, U = 15, p = 0.08, Mann-Whitney U test), Shannon (n = 8 each, U = 19, p = 0.19, Mann-Whitney U test).

(i) Histogram of genus-level taxa with p-values <0.05 evaluated by LEfSe analysis were visualized (n = 8 each).

Mean ± SEM; ^★ indicates p < 0.05; KO, knockout; WT, wild-type.