Multi-Omics Analysis for Assessment of Hepatoprotective Activity of Nature-derived Polyphenols in Case of Non-Alcoholic Fatty Liver Disease (NAFLD) Mouse Model

The aim of this study was to examine the hepatoprotective activity of multicomponent mixtures of natural origin in the BALB/C mouse model (59 female mice), with non-alcoholic fatty liver disease (NAFLD) induced by the administration of streptozotocin (STZ) in combination with a high-fat, high-fructose diet. The hepatoprotective activity of activated hydrolytic lignin (BP-Cx-1), metanolic fraction of BP-Cx-1 (BP-Cx-M) and isoflavones from kudzu Pueraria lobata roots (IFL) was evaluated using mass spectrometry (MS)-based omics technologies. Untargeted label-free DIA quantitation resulted in 8088 protein groups identification (FDR 1%) in 40 liver tissue extracts. All treatment NAFLD groups were closer to the control samples on the liver proteomic landscape with the best results shown for BP-Cx-1-m and ISF groups. This corroborated well with metabolomic fingerprinting by FTICR MS. In order to identify differences between specific groups, we applied the post-hoc Dunn’s Test and used Hedges' g as the Effect Size metric and 64 proteins that tended to return to their normal level after treatment were revealed. Pathophysiologically specific proteomics changes occurring during NAFLD were mostly associated with metabolism of the proteins (PSMD7, HCFC1) and deubiquitination pathways (UCHL3). It is worth noting that BP-Cx-1 isolated methanol fraction (Bp-Cx-m) demonstrated the pronounced increased hepatoprotective activity due to the enrichment with active components such as polyphenols. Additionally, it was shown that treatments with Bp-Cx-M and IFL significantly reduced DNA damage by 50% compared to the NAFLD untreated group. Finally, MS-based multiomics approach demonstrated its power as an advanced screening method for the assessment of the hepatoprotective activity and molecular mechanisms of action of complex multicomponent mixtures of natural origin.