Non-Targeted Metabolomics Reveal Apomorphine’s Therapeutic Effects and Lysophospholipid Alterations in Steatohepatitis

Figures

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  Figure 1. Partial improvement of adiposity and serum transaminase levels in PTEN KO mice by Apo administration. (A) Representative photos of H&E staining of the liver are shown. Control, PTEN KO, and Apo-treated PTEN KO mice are represented as normal, pathological, and treated states, respectively. (B) Comparison of MASH-related parameters across three groups, highlighting significant differences among three groups: (1, red) normal, (2, green) pathological, and (3, blue) treated. Yellow dots on the box and whiskers diagram represent the mean value and black dots indicate data points. ** p < 0.01, *** p < 0.001; ns, not significant.

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  Figure 2. Multivariate statistical analysis of liver metabolite data generated by non-targeted metabolomics. (A) Score plot of a sparse partial least squares-discriminant analysis (sPLS-DA) with positive (A,C) or negative (B,D) ion data. Liver extracts from normal, pathological, and treated mice were analyzed by LC-QTOF-MS. All peaks detected were submitted for sPLS-DA (A,B). Only peaks that were searched against the metabolite databases and then assigned to known compounds were submitted to sPLS-DA (C,D). Furthermore, only peaks that were confirmed as lysophospholipids (LPLs) or fatty acids by their MS2 spectra and retention times were submitted to sPLS-DA. The resulting score plot (E) and loading plot (F) are shown. LPC, lysophosphatidylcholine; LPE, lysophosphatidylethanolamine; LPI, lysophosphatidylinositol; FA, fatty acid; 16:0, palmitic acid; 18:0, stearic acid; 18:1, oleic acid; 20:0, eicosanoic acid; 20:1, eicosenoic acid; 20:3, eicosatrienoic acid; 20:4, arachidonic acid.

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  Figure 3. Box-plots of LPLs and fatty acids in liver extracts. These metabolites were evaluated as significant on multivariate analysis of non-targeted metabolomics. Detected amounts (peak heights) normalized by the protein amount were compared among three groups: (1, red) normal, (2, green) pathological, and (3, blue) treated. Yellow dots on the box and whiskers diagram represent the mean value and black dots indicate data points. * p < 0.05, ** p < 0.01, *** p < 0.001; ns, not significant; LPC, lysophosphatidylcholine; LPE, lysophosphatidylethanolamine; LPI, lysophosphatidylinositol; FA, fatty acid; 16:0, palmitic acid; 16:1, palmitoleic acid; 18:0, stearic acid; 18:2, linoleic acid; 20:3, eicosatrienoic acid; 20:4, arachidonic acid; [M + H]+, protonated ion; [M − H]−, deprotonated ion; [M + HCOO]−, formate adduct ion.

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  Figure 4. Box-plots of LPLs and other metabolites measured by targeted analysis. Detected amounts (peak areas) normalized by the protein amount were compared among three groups: (1, red) normal, (2, green) pathological, and (3, blue) treated. Yellow dots on the box and whiskers diagram represent the mean value and black dots indicate data points. * p < 0.05, ** p < 0.01, *** p < 0.001; ns, not significant; LPC, lysophosphatidylcholine; LPE, lysophosphatidylethanolamine; LPI, lysophosphatidylinositol; FA, fatty acid; DG, diacylglycerol; TG, triacylglycerol; 16:0, palmitic acid; 18:0, stearic acid; 18:1, oleic acid; 18:2, linoleic acid; 20:3, eicosatrienoic acid; 20:4, arachidonic acid; 22:6, docosahexaenoic acid; O1, mono-oxidized form.