NOVEL SERBIAN WHITE SOFT CHEESE IMPROVE GUT EPITHELIAL BARRIER AND AUTOPHAGY IN VITRO

Abstract

Objective: The application of natural starter cultures in fermented dairy products is gaining momentum due to their dual role in improving product quality and promoting host health. This study examines the potential of novel natural starter cultures used in the production of white soft cheese to modulate gut epithelial barrier function and autophagy in vitro. Methods: White soft cheese was produced using two artisanal starter strains: Lactococcus lactis subsp. lactis BGTRK4-21 and Lactobacillus plantarum BGGO7-29, both isolated from traditional Serbian dairy products. In vitro digestion of cheese samples aged 1, 10, 20, and 30 days was incubated with Caco-2 cells, and the expression of tight junction proteins, autophagy-related genes, as well as genes involved in antioxidant defense and antimicrobial peptide production, was quantified using RT-qPCR. Results: Exposure to digested cheeses significantly upregulated the expression of CLDN4, OCLN, BECN1, MAP1LC3B, p62, HBD1, SOD2, and GSTH genes in a time-dependent manner, with the most pronounced effects observed in 10- and 20-day-old cheese samples. These results suggest enhanced intestinal barrier integrity, activation of autophagy pathways, and improved cellular antioxidant and antimicrobial defense mechanisms. Conclusions: White soft cheese produced with selected natural starter cultures exerts beneficial effects on intestinal epithelial cells, indicating its potential as a functional food. These findings support the development of traditional dairy products with added probiotic-like benefits, contributing to gut health through barrier enhancement and autophagy modulation