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NF-κB-dependent anti-inflammatory activity of urolithins, gut microbiota ellagic acid-derived metabolites, in human colonic fibroblasts

  • Antonio González-Sarrías (a1), Mar Larrosa (a1), Francisco Abraham Tomás-Barberán (a1), Piero Dolara (a2) and Juan Carlos Espín (a1)...

Previous studies have reported the anti-inflammatory properties of pomegranate extracts, suggesting that ellagitannins (ET) and ellagic acid (EA) are the main anti-inflammatory compounds. However, both ET and EA are metabolised in vivo by the gut microbiota to yield urolithins (Uro) which can be found in the gut and in systemic bloodstream. The present study was carried out to evaluate the individual effect of EA and their microbiota-derived metabolites Uro on colon fibroblasts upon IL-1β treatment as an in vitro inflammation model. Uro-A and Uro-B (10 μm) inhibited PGE2 production (85 and 40 %, respectively) after IL-1β stimulation, whereas EA did not show any effect. Uro-A, but not Uro-B, down-regulated cyclo-oxygenase-2 (COX-2) and microsomal PGE synthase-1 (mPGES-1) mRNA expression and protein levels. Both Uro inhibited NF-κB translocation to nucleus. Slight but significant effects were found in the activation of mitogen-activated protein kinase (MAPK) pathways. Uro-A lowered c-Jun N-terminal kinase phosphorylation state, and both Uro inhibited p38 activation. No metabolites derived from Uro or EA were found in the cell media upon incubation of EA or Uro with the cells, and only traces of the compounds were found inside the cells. The present results suggest that Uro, mainly Uro-A, are the main compounds that are responsible for the pomegranate anti-inflammatory properties. The mechanism of action implicated seems to be via the inhibition of activation of NF-κB and MAPK, down-regulation of COX-2 and mPGES-1 expressions, and consequently,via the reduction of PGE2 production. Taking into account that Uro did not enter the cells, a competitive binding for IL-1β membrane receptor cannot be discarded.

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      NF-κB-dependent anti-inflammatory activity of urolithins, gut microbiota ellagic acid-derived metabolites, in human colonic fibroblasts
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*Corresponding author: Mar Larrosa, fax +34 968 396213, email
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1Rubio, CA & Befrits, R (2008) Colorectal cancer in Crohn's disease review of a 56-year experience in Karolinska Institute University Hospital. J Environ Pathol Toxicol 27, 257266.
2Rask, K, Zhu, Y, Wang, W, et al. (2006) Ovarian epithelial cancer: a role for PGE2-synthesis and signalling in malignant transformation and progression. Mol Cancer 5, 62.
3Wang, HW, Hsueh, CT, Lin, CF, et al. (2006) Clinical implications of microsomal prostaglandin E synthase-1 overexpression in human non-small-cell lung cancer. Ann Surg Oncol 13, 12241234.
4Surh, YJ, Chun, KS, Cha, HH, et al. (2001) Molecular mechanisms underlying chemopreventive activities of anti-inflammatory phytochemicals: down-regulation of COX-2 and iNOS through suppression of NF-kappa B activation. Mutat Res 1, 480481, 243–268.
5Catley, MC, Chivers, JE, Cambridge, LM, et al. (2003) IL-1beta-dependent activation of NF-kappaB mediates PGE2 release via the expression of cyclooxygenase-2 and microsomal prostaglandin E synthase. FEBS Lett 547, 7579.
6Masuko-Hongo, K, Berenbaum, F, Humbert, L, et al. (2004) Up-regulation of microsomal prostaglandin E synthase 1 in osteoarthritic human cartilage: critical roles of the ERK-1/2 and p38 signaling pathways. Arthritis Rheum 50, 28292838.
7Russel, MG, Engels, LG, Muris, JW, et al. (1998) Modern life's in the epidemiology of inflammatory bowel disease: a case–control study with special emphasis on nutritional factors. Eur J Gastroenterol Hepatol 10, 243249.
8Lansky, EP & Newman, RA (2007) Punica granatum (pomegranate) and its potential for prevention and treatment of inflammation and cancer. J Ethnopharmacol 109, 177206.
9Larrosa, M, Tomás-Barberán, FA & Espín, JC (2006) The dietary hydrolysable tannin punicalagin releases ellagic acid that induces apoptosis in human colon adenocarcinoma Caco-2 cells by using the mitochondrial pathway. J Nutr Biochem 17, 611625.
10Shukla, M, Gupta, K, Rasheed, Z, et al. (2008) Consumption of hydrolyzable tannins-rich pomegranate extract suppresses inflammation and joint damage in rheumatoid arthritis. Nutrition 24, 733743.
11Shukla, M, Gupta, K, Rasheed, Z, et al. (2008) Bioavailable constituents/metabolites of pomegranate (Punica granatum L.) preferentially inhibit COX-2 activity ex vivo and IL-1beta-induced PGE2 production in human chondrocytes in vitro. J Inflamm (Lond) 13, 59.
12Papoutsi, Z, Kassi, E, Chinou, I, et al. (2008) Walnut extract (Juglans regia L.) and its component ellagic acid exhibit anti-inflammatory activity in human aorta endothelial cells and osteoblastic activity in the cell line KS483. Br J Nutr 99, 715722.
13Cerdá, B, Cerón, JJ, Tomás-Barberán, FA, et al. (2003) Repeated oral administration of high doses of the pomegranate ellagitannin punicalagin to rats for 37 days is not toxic. J Agric Food Chem 51, 34933501.
14Cerdá, B, Espín, JC, Parra, S, et al. (2004) The potent in vitro antioxidant ellagitannins from pomegranate juice are metabolised into bioavailable but poor antioxidant hydroxy-6H-dibenzopyran-6-one derivatives by the colonic microbiota of healthy humans. Eur J Nutr 43, 205220.
15Cerdá, B, Periago, P, Espín, JC, et al. (2005) Identification of urolithin A as a metabolite produced by human colon microflora from ellagic acid and related compounds. J Agric Food Chem 53, 55715576.
16González-Sarrías, A, Azorín-Ortuño, M, Yáñez-Gascón, MJ, et al. (2009) Dissimilar in vitro and in vivo effects of ellagic acid and its microbiota-derived metabolites, urolithins, on the cytochrome P450 1A1. J Agric Food Chem 57, 56235632.
17Larrosa, M, González-Sarrías, A, Yañéz-Gascón, MJ, et al. (2010) Anti-inflammatory properties of a pomegranate extract and its metabolite urolithin-A in a colitis rat model and the effect of colon inflammation on the phenolic metabolism. J Nutr Biochem (In the Press).
18Morgan, DM (1998) Tetrazolium (MTT) assay for cellular viability and activity. Methods Mol Biol 79, 179183.
19Larrosa, M, Luceri, C, Vivoli, E, et al. (2009) Polyphenol metabolites from colonic microbiota exert anti-inflammatory activity on experimental inflammation models. Mol Nutr Food Res 53, 10441054.
20Jang, M, Cai, L, Udeani, GO, et al. (1997) Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science 275, 218220.
21Subbaramaiah, K, Chung, WJ, Michaluart, P, et al. (1998) Resveratrol inhibits cyclooxygenase-2 transcription and activity in phorbol ester-treated human mammary epithelial cells. J Biol Chem 273, 2187521882.
22Oh, SH & Lee, BH (2003) Induction of apoptosis in human hepatoblastoma cells by tetrandrine via caspase-dependent Bid cleavage and cytochrome c release. Biochem Pharmacol 66, 725731.
23Larrosa, M, González-Sarrías, A, García-Conesa, MT, et al. (2006) Urolithins, ellagic acid-derived metabolites produced by human colonic microflora, exhibit estrogenic and antiestrogenic activities. J Agric Food Chem 54, 16111620.
24Cerdá, B, Tomás-Barberán, FA & Espín, JC (2005) Metabolism of antioxidant and chemopreventive ellagitannins from strawberries, raspberries, walnuts, and oak-aged wine in humans: identification of biomarkers and individual variability. J Agric Food Chem 53, 227235.
25Faria, A, Monteiro, R, Mateus, N, et al. (2007) Effect of pomegranate (Punica granatum) juice intake on hepatic oxidative stress. Eur J Nutr 46, 271278.
26Adams, LS, Seeram, NP, Aggarwal, BB, et al. (2006) Pomegranate juice, total pomegranate ellagitannins, and punicalagin suppress inflammatory cell signalling in colon cancer cells. J Agric Food Chem 54, 980985.
27Cerdá, B, Soto, C, Albaladejo, MD, et al. (2006) Pomegranate juice supplementation in chronic obstructive pulmonary disease: a 5-week randomized, double-blind, placebo-controlled trial. Eur J Clin Nutr 60, 245253.
28Espín, JC, González-Barrio, R, Cerdá, B, et al. (2007) Iberian pig as a model to clarify obscure points in the bioavailability and metabolism of ellagitannins in humans. J Agric Food Chem 55, 1047610485.
29Mahida, YR, Beltinger, J, Makh, S, et al. (1997) Adult human colonic subepithelial myofibroblasts express extracellular matrix proteins and cyclooxygenase-1 and -2. Am J Physiol 273, G1341G1348.
30Mifflin, RC, Saada, JI, Di Mari, JF, et al. (2002) Regulation of COX-2 expression in human intestinal myofibroblasts: mechanisms of IL-1-mediated induction. Am J Physiol Cell Physiol 282, C824C834.
31Zhang, Y, DeWitt, DL, Murugesan, S, et al. (2004) Novel lipid-peroxidation- and cyclooxygenase-inhibitory tannins from Picrorhiza kurroa seeds. Chem Biodivers 1, 426441.
32Sadhu, SK, Okuyama, E, Fujimoto, H, et al. (2006) Prostaglandin inhibitory and antioxidant components of Cistus laurifolius, a Turkish medicinal plant. J Ethnopharmacol 108, 371378.
33Chell, S, Kaidi, A, Williams, AC, et al. (2006) Mediators of PGE2 synthesis and signalling downstream of COX-2 represent potential targets for the prevention/treatment of colorectal cancer. Biochim Biophys Acta 1766, 104119.
34Afaq, F, Malik, A, Syed, D, et al. (2005) Pomegranate fruit extract modulates UV-B-mediated phosphorylation of mitogen-activated protein kinases and activation of nuclear factor kappa B in normal human epidermal keratinocytes paragraph sign. Photochem Photobiol 81, 3845.
35Newton, R, Kuitert, LM, Bergmann, M, et al. (1997) Evidence for involvement of NF-kappaB in the transcriptional control of COX-2 gene expression by IL-1beta. Biochem Biophys Res Commun 237, 2832.
36Guan, Z, Buckman, SY, Miller, BW, et al. (1998) Interleukin-1beta-induced cyclooxygenase-2 expression requires activation of both c-Jun NH2-terminal kinase and p38 MAPK signal pathways in rat renal mesangial cells. J Biol Chem 273, 2867028676.
37LaPointe, MC & Isenović, E (1999) Interleukin-1β regulation of inducible nitric oxide synthase and cyclooxygenase-2 involves the p42/44 and p38 MAPK signaling pathways in cardiac myocytes. Hypertension 33, 276282.
38Yang, CM, Chien, CS, Hsiao, LD, et al. (2002) Interleukin-1beta-induced cyclooxygenase-2 expression is mediated through activation of p42/44 and p38 MAPKS, and NF-kappaB pathways in canine tracheal smooth muscle cells. Cell Signal 14, 899911.
39González-Sarrías, A, Espín, JC, Tomás-Barberán, FA, et al. (2009) Gene expression, cell cycle arrest and MAPK signalling regulation in Caco-2 cells exposed to ellagic acid and its metabolites, urolithins. Mol Nutr Food Res 53, 686698.
40Masamune, A, Satoh, M, Kikuta, K, et al. (2005) Ellagic acid blocks activation of pancreatic stellate cells. Biochem Pharmacol 70, 869878.
41Krysan, K, Reckamp, KL, Dalwadi, H, et al. (2005) Prostaglandin E2 activates mitogen-activated protein kinase/Erk pathway signaling and cell proliferation in non-small cell lung cancer cells in an epidermal growth factor receptor-independent manner. Cancer Res 65, 62756281.
42Sirois, J & Richards, JS (1993) Transcriptional regulation of the rat prostaglandin endoperoxide synthase 2 gene in granulosa cells. Evidence for the role of a cis-acting C/EBP beta promoter element. J Biol Chem 268, 2193121938.
43Inoue, H, Yokoyama, C, Hara, S, et al. (1995) Transcriptional regulation of human prostaglandin-endoperoxide synthase-2 gene by lipopolysaccharide and phorbol ester in vascular endothelial cells. Involvement of both nuclear factor for interleukin-6 expression site and cAMP response element. J Biol Chem 270, 2496524971.
44Subbaramaiah, K, Yoshimatsu, K, Scherl, E, et al. (2004) Microsomal prostaglandin E synthase-1 is overexpressed in inflammatory bowel disease. Evidence for involvement of the transcription factor Egr-1. J Biol Chem 279, 1264712658.
45Pascual, RM, Carr, EM, Seeds, MC, et al. (2006) Regulatory features of interleukin-1beta-mediated prostaglandin E2 synthesis in airway smooth muscle. Am J Physiol Lung Cell Mol Physiol 290, L501L508.
46Meadows, AL, Kong, B, Berdichevsky, M, et al. (2008) Metabolic and morphological differences between rapidly proliferating cancerous and normal breast epithelial cells. Biotechnol Prog 24, 334341.
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