GSE75239
Microarray analysis profiling of the effect of cowpea phenolic extract on gene expression in bovine peripheral blood in vitro
Plant-derived secondary metabolites found in animal feed sources are beneficial for nutrition and health. Cowpea is a protein and phenolic-rich forage used as feed resource in animal system. The objective of this study was to understand the effect of cowpea secondary metabolites on gene expression in cows blood in vitro. Whole blood collected from Holstein Friesian cows were treated with 10 ug/ml of cowpea leaf phenolic extract and untreated samples served as control. Total RNA was isolated and pooled together for microarray analysis. The Agilent one color bovine (v2) 4x44K array was used and preliminary gene expression profiles generated using Cy3 labeled cDNA from CPE-stimulated and untreated samples. Gene expression analysis revealed a total of 3170 differentially expressed genes- 1716 up regulated and 1454 down regulated genes respectively. Pathway analysis identified CPE treatment association with innate immune response pathways including Toll-like receptor (TLR) signaling pathway, the Wnt signaling pathway, inflammation response pathway, and increased expression of the transcription factor NFKB1 were observed. Treatment with CPE decreased the expression of proinflammatory cytokines IL1A, IL1B and IL21. Quantitative real time PCR was performed to validate some gene members of the Toll-like receptor, inflammation response and Wnt signaling pathways. In vitro treatment with CPE impacted global gene expression profile in cow blood and results obtained in this study shows the potential immuno-modulatory properties of cowpea feed phenolic in cows. Overall design: The global gene expression of the effect of cowpea phenolic extract (CPE) was measured in bovine peripheral blood.The experiment involved two groups; cowpea phenolic extract (CPE) treated samples vs untreated control group. Pooled RNA samples from each group was hybridized on Agilent one color bovine (v2) 4x44K microarray slides. Two slides were prepared each with 4 array compartment.