Isomer-Specific Effects of cis-9,trans-11- and trans-10,cis-12-CLA on Immune Regulation in Ruminal Epithelial Cells

SIMPLE SUMMARY: The significant contribution of rumen microbiota to the balance of the innate immunity of rumen epithelium has been extensively verified. As the natural rumen microbial metabolites, information regarding the immunoprotective effects of different conjugated linoleic acid (CLA) isomers on ruminal epithelial cells (RECs) is limited. In this study, the 100 μM trans-10,cis-12-CLA exerted better anti-inflammatory effects than the cis-9,trans-11-CLA by significantly downregulating the expression of genes related to inflammation, cell proliferation and migration in RECs upon lipopolysaccharide (LPS) stimulation. The trans-10,cis-12-CLA, but not cis-9,trans-11-CLA, significantly suppressed the biological signals of gene ontology (GO) terms’ response to lipopolysaccharide, the regulation of signal transduction and cytokine production and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways NF-κB, chemokine, NOD-like receptor, Hippo, PI3K-Akt, TGF-β and Rap1 signaling in RECs upon LPS stimulation. Furthermore, pretreatment with trans-10,cis-12-CLA significantly reduced the expression of lipogenic genes and the biosynthesis of the unsaturated fatty acid pathway in RECs compared with the LPS group, however, cis-9,trans-11-CLA exhibited the opposite results. These results suggest the distinct isomer differences of CLA in the regulation of inflammatory responses and adipocytokine signaling in RECs and will provide important references for determining their target use in the future. ABSTRACT: In this study, we used transcriptomics and qPCR to investigate the potential immunoprotective effects of different conjugated linoleic acid (CLA) isomers, the natural rumen microbial metabolites, on lipopolysaccharide (LPS)-induced inflammation of ruminal epithelial cells (RECs) in vitro. The results showed that 100 μM trans-10,cis-12-CLA exerted higher anti-inflammatory effects than cis-9,trans-11-CLA by significantly downregulating the expression of genes related to inflammation, cell proliferation and migration in RECs upon LPS stimulation. Transcriptomic analyses further indicated that pretreatment with trans-10,cis-12-CLA, but not cis-9,trans-11-CLA, significantly suppressed the biological signals of GO terms’ response to LPS, the regulation of signal transduction and cytokine production and KEGG pathways NF-κB, chemokine, NOD-like receptor, Hippo, PI3K-Akt, TGF-β and Rap1 signaling in RECs upon LPS stimulation. Furthermore, pretreatment with trans-10,cis-12-CLA significantly reduced the expression of lipogenic genes and the biosynthesis of the unsaturated fatty acid pathway in RECs compared with the LPS group, however, cis-9,trans-11-CLA exhibited the opposite results. These results suggest the distinct isomer differences of CLA in the regulation of inflammatory responses and adipocytokine signaling in RECs and will provide important references for determining their target use in the future.