Liver steatosis can evolve to steatohepatitis (NASH) through a series of biochemical steps related to oxidative stress in hepatocytes. Antioxidants, such as silybin, have been proposed as a treatment of patients with nonalcoholic fatty liver disease (NAFLD) and NASH. In this study, we evaluated, in patients with histologically documented NASH, the oxidant/antioxidant status and lipid “fingerprint” in the serum of NASH patients, both in basal conditions and after 12 months of treatment with silybin-based food integrator Realsil (RA). The oxidant/antioxidant status analysis showed the presence of a group of patients with higher basal severity of disease (NAS scores 4.67 ± 2.5) and a second group corresponding to borderline NASH (NAS scores = 3.8 ± 1.5). The chronic treatment with RA changed the NAS score in both groups that reached the statistical significance only in group 2, in which there was also a significant decrease of serum lipid peroxidation. The lipidomic profile showed a lipid composition similar to that of healthy subjects with a restoration of the values of free cholesterol, lysoPC, SM, and PC only in group 2 of patients after treatment with RA. Conclusion. These data suggest that lipidomic and/or oxidative status of serum from patients with NASH could be useful as prognostic markers of response to an antioxidant treatment.

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Bioactive peptides are present in a latent state, encrypted within the amino acid sequence of milk proteins, requiring enzymatic proteolysis for their release. They can be produced by gastrointestinal digestion or food processing, thus they can be present in fermented milks, cheese and also in the by-products of dairy industry such as waste whey. The spectrum of biological activity covered by milk-derived peptides is extremely wide, including antibacterial, immunostimulating, antihypertensive, antithrombotic and opioid actions. However, the characterisation of milk-derived peptides with classical analytical methodologies is severely challenged by the complexity of the milk protein fraction and by the wide dynamic range of relative peptide abundance in both dairy products and by-products. Here we report the characterisation of the peptide fraction released in the whey during the different production stages of Mozzarella di Bufala Campana cheese. The peptide extracts were separated by RP HPLC and analysed by MS in order to identify the peptides produced and to trace the pathway of formation of potential bioactive peptides. The antioxidant properties and the modulatory effect on the cell cycle exerted by the peptide extracts were also studied in CaCo2 cell line. We found that a significant antiproliferative effect on CaCo2 was exerted by Mozzarella di Bufala waste whey peptides.

The structural characterization of the active peptide fraction and the elucidation of the effects induced by its treatment on H-CaCo2 cells in vitro demonstrated an activity of this peptide sub-fraction in the modulation of cell cycle, thus suggesting potential application for the development of nutraceuticals as well as health-promoting functional foods.