Untargeted serum metabolites profiling in high-fat diet mice supplemented with enhanced palm tocotrienol-rich fraction using UHPLC-MS

Abstract

Excessive high fat dietary intake promotes risk of developing non-alcoholic fatty liver disease (NAFLD) and predisposed with oxidative stress. Palm based tocotrienol-rich fraction (TRF) has been reported able to ameliorate oxidative stress but exhibited poor bioavailability. Thus, we investigated whether an enhanced formulation of TRF in combination with palm kernel oil (medium-chain triglycerides) (ETRF) could ameliorate the effect of high-fat diet (HFD) on leptin-deficient male mice. All the animals were divided into HFD only (HFD group), HFD supplemented with ETRF (ETRF group) and HFD supplemented with TRF (TRF group) and HFD supplemented with PKO (PKO group). After 6 weeks, sera were collected for untargeted metabolite profiling using UHPLC-Orbitrap MS. Univariate analysis unveiled alternation in metabolites for bile acids, amino acids, fatty acids, sphingolipids, and alkaloids. Bile acids, lysine, arachidonic acid, and sphingolipids were downregulated while xanthine and hypoxanthine were upregulated in TRF and ETRF group. The regulation of these metabolites suggests that ETRF may promote better fatty acid oxidation, reduce oxidative stress and pro-inflammatory metabolites and acts as anti-inflammatory in fatty liver compared to TRF. Metabolites regulated by ETRF also provide insight of its role in fatty liver. However, further investigation is warranted to identify the mechanisms involved.

Untargeted muscle tissue metabolites profiling in young, adult, and old rats supplemented with tocotrienol-rich fraction

“…When the samples were ready to run, it was reconstituted with 100 ul isopropanol:MeOH (1:1) and 100 ul was transferred through a filter to vials using syringe and syringe filters. For the quality control (QC) sample, 5 uL of all samples were mixed from each vial into a tube, spun down, and 100 ul were transferred into a vial and capped (Goon et al, 2021). The vials were randomly placed into the Liquid Chromatography-Mass Spectrometry and run for 3 days.…”

Tocotrienol in the Management of Nonalcoholic Fatty Liver Disease: A Systematic Review

“…Another genetically modified mouse model used was B6.Cg-LepOb/J mice, which develop metabolic syndrome and obesity spontaneously. They were fed a high-fat diet for 6 weeks to induce NAFLD [ 41 ]. The T3 treatments and their durations have been summarised in Table 1 .…”

“…FXR was shown to suppress steatosis by inhibiting SREBP1c and induction on PPARα and its target, CPT1 [ 68 ]. Palm TRF (unformulated or enhanced) was shown to increase the protein and mRNA expression of FXR in the liver [ 41 ]. However, no significant changes in mRNA expression of FXR targets, i.e., small heterodimer partner and signal transducer and activator of transcription 3, were observed between treated and untreated B6.Cg-LepOb/J male fed with a high-fat diet [ 41 ].…”

“…Palm TRF (unformulated or enhanced) was shown to increase the protein and mRNA expression of FXR in the liver [ 41 ]. However, no significant changes in mRNA expression of FXR targets, i.e., small heterodimer partner and signal transducer and activator of transcription 3, were observed between treated and untreated B6.Cg-LepOb/J male fed with a high-fat diet [ 41 ]. The authors postulated that the fatty acids in the vehicle for T3 could attenuate FXR signalling despite the increased FXR expression [ 41 ].…”

Serum Metabolomic Alteration in Rats with Osteoarthritis Treated with Palm Tocotrienol-Rich Fraction Alone or in Combination with Glucosamine Sulphate

“…For MS/MS scans, a resolution of 15,000 was applied along with stepped normalised collision energy settings of 20, 40, and 60 AU. These settings were in accordance with those described by Goon et al [67].…”