Modulation of barrier integrity and inflammation in the caecum of broiler chickens exposed to glycerides blend

The integrity of the intestinal barrier and the regulation of local inflammatory responses are crucial for gut homeostasis and resistance in broiler chickens under intensive farming and reduced antibiotic use. This study aimed to evaluate the effects of a glycerides blend on both the expression and protein-level regulation of barrier and inflammation markers in the caecum of broiler chickens. The study was conducted on an industrial farm using Cobb 500 broiler chickens. Gut tissue samples were collected at 2, 3, 4, 5, and 6 weeks of age. Occludin and TNF-α expression was assessed by quantitative PCR, while protein content was detected using Western blot analysis. The correlation between mRNA and protein levels was analyzed, and an integrative barrier-inflammation balance index (BIBI) was calculated to identify functional shifts in gut homeostasis. The results demonstrated that glyceride blend supplementation induced significant age-dependent and phase-specific modulation of intestinal functioning. In contrast to the control group, which exhibited relatively stable transcriptional profiles, the treatment with the glycerides blend resulted in a significant upregulation of Occludin mRNA and a sustained increase in Occludin protein content beginning from the second week of exposure (P ≤ 0.001). Glycerides blend supplementation caused a persistent reduction in TNF-α protein content compared to the control. In spite of the decrease in TNF-α protein, a delayed increase in TNF-α mRNA expression was observed at the 6th week (P ≤ 0.001). Correlation analysis has shown an incomplete and variable dependence between mRNA and protein levels. The obtained data support the concept of multiple regulatory pathways controlling both intestinal barrier and immunity. The BIBI analysis confirmed a clear shift (P ≤ 0.001) toward barrier-oriented regulation and attenuation of inflammation in the caecum of glyceride-exposed chickens. These findings indicate that the glycerides blend promotes adaptive reprogramming of intercellular adhesion and inflammatory balance, improves gut barrier integrity, and restricts TNF-α-mediated injury. Considering the observed beneficial effects, applied feed additive demonstrates promising features as at least a part of an antibiotic-free strategy for industrial poultry farming.