Abstract
The present study was performed to investigate the anti-inflammatory potential of a 116-kDa glycoprotein isolated from Ulmus davidiana Nakai (UDN glycoprotein) in lipopolysaccaride (LPS)-treated cancerous human colon epithelial cells (HT-29 cells). UDN glycoprotein inhibited the production of intracellular superoxide anion (O ·−2 ), hydrogen peroxides (H2O2), and nitric oxide (NO), whereas normalized the activity of anti-oxidant enzymes [superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX)], accompanying the inhibition of manganese-superoxide dismutases (Mn-SOD) activity in LPS-treated HT-29 cells. In addition, UDN glycoprotein blocked the DNA binding activity of activator protein-1 (AP-1) through suppression of c-Jun and c-Fos activities, respectively. We also evaluated the anti-inflammatory potential of UDN glycoprotein based on the activity of the pro-inflammatory signal mediators [inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and matrix metalloproteinases-9 (MMP-9)]. The results showed that UDN glycoprotein (200 μg/ml) has an inhibitory effect on the activation of iNOS, COX-2, and MMP-9 proteins in the LPS-treated HT-29 cells. From these results, we suggest that UDN glycoprotein is one of the potential anti-inflammatory agents that blocks LPS-mediated inflammatory signal pathway in HT-29 cells. Here, we speculate that UDN glycoprotein could be used as an antioxidative agent for inflammatory gastrointestinal cancers.
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References
Haller D, Jobin C (2004) Interaction between resident luminal bacteria and the host: can a healthy relationship turn sour? J Pediatr Gastroenterol Nutr 38:123–136
Bhattacharyya J, Biswas S, Datta AG (2004) Mode of action of endotoxin: role of free radicals and antioxidants. Curr Med Chem 11:359–368
Kim JS, Narula AS, Jobin C (2005) Salvia miltiorrhiza water-soluble extract, but not its constituent salvianolic acid B, abrogates LPS-induced NF-kappaB signalling in intestinal epithelial cells. Clin Exp Immunol 141:288–297
Cario E, Rosenberg IM, Brandwein SL, Beck PL, Reinecker HC, Podolsky DK (2000) Lipopolysaccharide activates distinct signaling pathways in intestinal epithelial cell lines expressing Toll-like receptors. J Immunol 164:966–972
Gardiner KR, Halliday MI, Barclay GR, Milne L, Brown D, Stephens S, Maxwell RJ, Rowlands BJ (1995) Significance of systemic endotoxaemia in inflammatory bowel disease. Gut 36:897–901
Kruidenier L, Verspaget HW (2002) Oxidative stress as a pathogenic factor in inflammatory bowel disease—radicals or ridiculous? Aliment Pharm Ther 16:1997–2015
Kinnula VL, Crapo JD (2004) Superoxide dismutases in malignant cells and human tumors Free Radic Biol Med 36:718–744
Visner GA, Dougall WC, Wilson JM, Burr IA, Nick HS (1990) Regulation of manganese superoxide dismutase by lipopolysaccharide, interleukin-1, and tumor necrosis factor. Role in the acute inflammatory response J Biol Chem 265:2856–2864
Shull S, Heintz NH, Periasamy M, Manohar M, Janssen YM, Marsh JP, Mossman BT (1991) Differential regulation of antioxidant enzymes in response to oxidants J Biol Chem 266:24398–24403
Nelson KK, Melendez JA (2004) Mitochondrial redox control of matrix metalloproteinases Free Radic Biol Med 37:768–784
Kruidenier L, Kuiper I, van Duijn W, Marklund SL, van Hogezand RA, Lamers CB, Verspaget HW (2003) Differential mucosal expression of three superoxide dismutase isoforms in inflammatory bowel disease J Pathol 201:7–16
Surh YJ, Kundu JK, Na HK, Lee JS (2005) Redox-sensitive transcription factors as prime targets for chemoprevention with anti-inflammatory and antioxidative phytochemicals J Nutr 135:2993S–3001S
Eferl R, Wagner EF (2003) AP-1: a double edged sword in tumorigenesis. Nat Rev Cancer 3:859–868
Sen CK, Packer L (1996) Antioxidant and redox regulation of gene transcription. FASEB J 10:709–720
de la Lastra CA, Villegas I (2005) Resveratrol as an anti-inflammatory and anti-aging agent: mechanisms and clinical implications Mol Nutr Food Res 49:405–430
Mook OR, Frederiks WM, Van Noorden CJ (2004) The role of gelatinases in colorectal cancer progression and metastasis Biochim Biophys Acta 1705:69–89
Lee SJ, Heo KS, Oh PS, Lim K, Lim KT (2004) Glycoprotein isolated from Ulmus davidiana Nakai inhibits TPA-induced apoptosis through nuclear factor-kappa B in NIH/3T3 cells Toxicol Lett 146:159–174
Ko JH, Lee SJ, Lim KT (2005) 116 kDa glycoprotein isolated from Ulmus davidiana Nakai (UDN) inhibits glucose/glucose oxidase (G/GO)-induced apoptosis in BNL CL.2 cells J Ethnopharmacol 100:339–346
Lee SJ, Oh PS, Ko JH, Lim K, Lim KT (2006) Protective effect of glycoprotein isolated from Ulmus davidiana Nakai on carbon tetrachloride-induced mouse liver injury J Pharm Pharmacol 58:143–152
Suzuki M, Hisamatsu T, Podolsky DK (2003) Gamma interferon augments the intracellular pathway for lipopolysaccharide (LPS) recognition in human intestinal epithelial cells through coordinated up-regulation of LPS uptake and expression of the intracellular Toll-like receptor 4-MD-2 complex Infect Immun 71:3503–3511
Neville DM, Glossmann H (1974) Molecular weight determination of membrane protein and glycoprotein subunits by discontinuous gel electrophoresis in dodecyl sulfate Methods Enzymol 32:92–102
Royall JA, Ischiropoulos H (1993) Evaluation of 2′,7′-dichlorofluorescin and dihydrorhodamine 123 as fluorescent probes for intracellular H2O2 in cultured endothelial cells. Arch Biochem Biophys 302:348–355
Zhang Z, Leonard SS, Huang C, Vallyathan V, Castranova V, Shi X (2003) Role of reactive oxygen species and MAPKs in vanadate-induced G(2)/M phase arrest. Free Radic Biol Med 34:1333–1342
Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44: 276–287
Thomson JF, Nance SL, Tollaksen SL (1978) Spectrophotometric assay of catalase with perborate as substrate. Proc Soc Exp Biol Med 157: 33–35
Paglia DE, Valentine WN (1967) Studies on the quantitative and qualitative characterization of erythrocyte glutathine peroxidase. J Lab Clin Med 70: 158–169
Lee SJ, Oh PS, Ko JH, Lim K, Lim KT (2004) A 150-kDa glycoprotein isolated from Solanum nigrum L. has cytotoxic and apoptotic effects by inhibiting the effects of protein kinase C alpha, nuclear factor-kappa B and inducible nitric oxide in HCT-116 cells. Cancer Chemother Pharmacol 54: 562–572
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275
Lee SJ, Lim KT (2006) Apoptosis induced by glycoprotein (150-kDa) isolated from Solanum nigrum L. is not related to intracellular reactive oxygen species (ROS) in HCT-116 cells. Cancer Chemother Pharmacol 57:507–516
Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR (1982) Analysis of nitrate, nitrite, and [15N] nitrate in biological fluids. Anal Biochem 126:131–138
Fridovich I (1983) Superoxide radical: an endogenous toxicant. Annu Rev Pharmacol Toxicol 23:239–257
Lakari E, Paakko P, Kinnula VL (1998) Manganese superoxide dismutase, but not CuZn superoxide dismutase, is highly expressed in the granulomas of pulmonary sarcoidosis and extrinsic allergic alveolitis. Am J Respir Crit Care Med 158:589–596
Clerch LB, Massaro D (1993) Tolerance of rats to hyperoxia. Lung antioxidant enzyme gene expression J Clin Invest 91:499–508
Surh YJ, Chun KS, Cha HH, Han SS, Keum YS, Park KK, Lee SS (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 480–481:243–268
Crowell JA, Steele VE, Sigman CC, Fay JR (2003) Is inducible nitric oxide synthase a target for chemoprevention? Mol Cancer Ther 2:815–823
Hill KA, Wang KL, Stryker SJ, Gupta R, Weinrach DM, Rao MS (2004) Comparative analysis of cell adhesion molecules, cell cycle regulatory proteins, mismatch repair genes, cyclooxygenase-2, and DPC4 in carcinomas arising in inflammatory bowel disease and sporadic colon cancer. Oncol Rep 11:951–956
Bachschmid M, Schildknecht S, Ullrich V (2005) Redox regulation of vascular prostanoid synthesis by the nitric oxide-superoxide system. Biochem Biophys Res Commun 338:536–542
Hinz B, Brune K (2002) Cyclooxygenase-2–10 years later. J Pharmacol Exp Ther 300:367–375
Kouzarides T, Ziff E (1988) The role of the leucine zipper in the fos-jun interaction. Nature 336:646–651
Naito Y, Yoshikawa T (2005) Role of matrix metalloproteinases in inflammatory bowel disease. Mol Aspects Med 26:379–390
Kim JH, Lee KW, Lee MW, Lee HJ, Kim SH, Surh YJ (2006) Hirsutenone inhibits phorbol ester-induced upregulation of COX-2 and MMP-9 in cultured human mammary epithelial cells: NF-kappaB as a potential molecular target. FEBS Lett 580:385–392
Warner RL, Bhagavathula N, Nerusu KC, Lateef H, Younkin E, Johnson KJ, Varani J (2004) Matrix metalloproteinases in acute inflammation: induction of MMP-3 and MMP-9 in fibroblasts and epithelial cells following exposure to pro-inflammatory mediators in vitro. Exp Mol Pathol 76:189–195
Grau R, Punzon C, Fresno M, Iniguez MA (2006) Peroxisome-proliferator-activated receptor alpha agonists inhibit cyclo-oxygenase 2 and vascular endothelial growth factor transcriptional activation in human colorectal carcinoma cells via inhibition of activator protein-1. Biochem J 395:81–88
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This work was supported by the Korea Research Foundation Grant funded by the Korean Government (MOEHRD) (KRF-2005-041-F00082).
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Lee, SJ., Lim, KT. UDN glycoprotein inhibits activator protein-1 and matrix metalloproteinase-9 via blocking of oxygen radicals in HT-29 cells. Mol Cell Biochem 304, 13–23 (2007). https://doi.org/10.1007/s11010-007-9481-y
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DOI: https://doi.org/10.1007/s11010-007-9481-y