Integrated multi-omics reveals the beneficial role of chlorogenic acid in improving the growth performance and immune function of immunologically stressed broilers

Intensive production can cause immunological stress in commercial broilers.Chlorogenic acid(CGA)regulates the intestinal microbiota,barrier function,and immune function in chickens.As complex interrelations regulate the dynamic interplay between gut microbiota,the host,and diverse health outcomes,the aim of this study was to elucidate the immunoregulatory mechanisms of CGA using multiomics approaches.A total of 240 one-day-old male broilers were assigned to a 2×2 factorial design with 2 CGA levels(0 or 500 mg/kg)either with or without dexamethasone(DEX)injection for a 21-day experimental period.Therefore,there were 4 dietary treatments:control,DEX,CGA,and DEX+CGA,with 6 replicates per treatment.CGA supplementation improved(P<0.05)growth performance,jejunal morphology,jejunal barrier function,and immune function in DEX-treated broilers.Moreover,in DEX+CGA-treated broilers,the increase in gut microbiome diversity(P<0.05)was consistent with a change in taxonomic composition,especially in the Clostridiales vadin BB60_group.Additionally,the levels of short-chain fatty acids increased remarkably(P<0.01)after CGA supplementation.This was consistent with the Kyoto Encyclopedia of Genes and Genomes analysis results that the“pyruvate fermentation to butanoate”pathway was more enriched(P<0.01)in the DEX+CGA group than in the DEX group.Proteomics revealed that CGA treatment increased the expression of several healthpromoting proteins,thymosin beta(TMSB4X)and legumain(LGMN),which were verified by multiple reaction monitoring.Metabolomics revealed that CGA treatment increased the expression of healthpromoting metabolites(2,4-dihydroxy benzoic acid and homogentisic acid).Proteomic and metabolic analyses showed that CGA treatment regulated the peroxisome proliferator-activated receptor(PPAR)and mitogen-activated protein kinase(MAPK)pathways.Western blotting results support these findings.Pearson’s correlation analyses showed correlations(P<0.01)between altered immune function,jejunal barrier function,different microbiota,proteins,and metabolites parameters.Overall,our data indicate that CGA treatment increased growth performance and improved the immunological functions of DEXtreated broilers by regulating gut microbiota and the PPAR and MAPK pathways.The results offer novel insights into a CGA-mediated improvement in immune function and intestinal health.