Journal: ACS Omega

Article Title: Polystyrene Microplastics Activate Noncanonical TGF‑β Signaling and Metabolomic Reprogramming to Promote Epithelial-Mesenchymal Transition and Fibrosis in the Kidney

doi: 10.1021/acsomega.5c10834

Figure Lengend Snippet: NMR-based metabolomic perturbation in NRK-52E cells after 96 h of PS-MPs treatment at low doses, 0.02, and high doses, 0.5 mg/mL. Control vs low doses: 0.02 mg/mL PS-MPs. (A) Representative 1 H NMR spectrum of metabolites extracted from control and PS-MPs (0.02 and 1 mg/mL) exposure. Control vs low doses: 0.02 mg/mL PS-MPs. (B) OPLS-DA of control (green color) and 0.02 mg/mL PS-MPs (red color). (C) Permutation test statistics with R2Y 0.99 and Q2 0.67. (D) The result of random forest classification with an out-of-bag (OOB) error of 0.33 and overall accuracy of 67% of control vs 0.02 mg/mL PS-MPs-treated groups. (E) VIP score plot. (F) Univariate analysis showing the significant metabolites in the control and PS-MPs (0.02 mg/mL)-treated group detected by 1 H NMR spectroscopy. The x -axis represents the metabolite, and the y -axis shows the relative concentration when control (represented by green color) and PS-MPs, 0.02 mg/mL (represented by red color), were compared. (G) Correlation heat map for the differential metabolites identified in the control vs PS-MPs (0.02 mg/mL) exposed group as calculated by z -scores. (H) Pathway analysis indicated an impact factor less than 0.1, thus not very significant. Control vs high doses: 0.5 mg/mL PS-MPs. (I) OPLS-DA of control (green color) and 0.5 mg/mL PS-MPs concentration (red color). (J) Permutation test statistics with R2Y 1 and Q2 0.59. (K) The result of random forest classification with an out-of-bag (OOB) error of 0.0 and an overall accuracy of 100% of control vs 0.5 mg/mL PS-MPs-treated groups (L) VIP score plot. (M) Univariate analysis showing the significant metabolites in the control and PS-MPs (0.5 mg/mL)-treated group detected by 1 H NMR spectroscopy. The x -axis represents the metabolite, and the y -axis represents the relative concentration when control (represented by green color) and PS-MPs (0.5 mg/mL, represented by red color) were compared. (N) Correlation heat map for the differential metabolites identified in control vs PS-MPs (0.5 mg/mL)-exposed group, as calculated by z-scores. (O) Pathway analysis showing an impact factor of more than 0.1 was considered. The top red one is phenylalanine, tyrosine, and tryptophan biosynthesis, and the other, with an impact factor between 0.2 and 0.4 is phenylalanine metabolism.

Article Snippet: Manual baseline correction (Whittaker method) was done with a Chenomx NMR processor (NMR Suite, v8.1, Chenomx Inc., Canada), and peak fitting of metabolites was done using the Chenomx profiler.

Techniques: Metabolomic, Control, Structural Proteomics, Concentration Assay