The objectives of this experiment were to compare the in-vitro anti-inflammatory activity of crude extracts (cold water (CWE), hot water and 80% ethanol extract) of F. vesiculosus and to predict the key molecular targets of the extract with most anti-inflammatory activity in-vitro. In TNF-alpha. challenged Caco-2 cells, of the three crude extracts, the CWE exhibited maximum inhibition of IL-8 production. In the ex-vivo challenged porcine colonic tissue, CWE inhibited the expression (>2 fold) of inflammatory mediators (PTGS2, C5, LYZ), cytokines (IL17A, IL8), chemokines (CCL2, CXCL2, CXCL10, CXCL11), cell adhesion molecules (ICAM1, VCAM1), toll like receptors (TLR4, TLR7) and components of NF-kappa B (NFKB1, RELB), MAPIC (MAP3K8) and AP-1 (CJUN) pathways. The gene expression analysis suggest that the CWE does contain immunomodulatory bioactive compound/s that mediated through the interferon regulatory system 2, TNF-alpha inducing protein 3 and TNF-a receptor 2 and thus has potential application for human and animal health. (C) 2016 Elsevier Ltd. All rights reserved.