Investigating the growth performance, meat quality, immune function and proteomic profiles of plasmal exosomes in Lactobacillus plantarum-treated broilers with immunological stress

Abstract

Exosomes are extracellular membranous nanovesicles that carry functional molecules to mediate cell-to-cell communication. To date, whether probiotics improve the immune function of broilers by plasmal exosome cargo is unclear. In this study, 300 broilers were allocated to three treatments: control diet (CON group), control diet + dexamethasone injection (DEX group), and control diet containing 1 × 10⁸ cfu g⁻¹ P8 + DEX injection (P8 + DEX group). The growth performance, meat quality and immune function of plasma and jejunal mucosa were detected. Exosomes were isolated from the plasma and characterized. Then, the exosome protein profile was determined by proteomic analysis. Correlation analyses between the exosomal proteins and growth performance, meat quality, immune function were performed. Lastly, the related protein levels were verified by multiple reaction monitoring (MRM). Results showed that P8 treatment increased the growth performance, meat quality and immune function of DEX-induced broilers with immunological stress. Moreover, the average diameters, cup-shaped morphology and expressed exosomal proteins confirmed that the isolated extracellular vesicles were exosomes. A total of 784 proteins were identified in the exosomes; among which, 126 differentially expressed proteins (DEPs) were found between the DEX and CON groups and 102 DEPs were found between the P8 + DEX and DEX groups. Gene ontology analysis indicated that DEPs between the DEX and CON groups are mainly involved in the metabolic process, cellular anatomical entity, cytoplasm, etc. DEPs between the P8 + DEX and DEX groups are mainly involved in the multicellular organismal process, response to stimulus, cytoplasm, etc. Pathway analysis revealed that most of the DEPs between the DEX and CON groups participated in the ECM–receptor interaction, focal adhesion, regulation of actin cytoskeleton, etc. Most of the DEPs between the P8 + DEX and DEX groups participated in the ErbB and PPAR signaling pathways. Moreover, many DEPs were correlated with the altered parameters of growth performance, meat quality and immunity in P8-treated broilers. MRM further revealed that the upregulated FABP6 and EPCAM in the DEX group were decreased by P8 + DEX treatment, and the downregulated C1QTNF3 in the DEX group was increased by P8 + DEX treatment. In conclusion, our findings demonstrated that P8 may promote the immune function, growth performance and meat quality of broilers with immunological stress by regulating the plasma exosomal proteins, especially the proteins of FABP6, EPCAM and C1QTNF3 and the pathway of PPAR (ILK/FABP6).