Lactobacillus GG restoration of the gliadin induced epithelial barrier disruption: the role of cellular polyamines

BMC Microbiology

Francesco Russo²  Valeria Tutino¹  Maria Notarnicola¹  Michele Linsalata²  Antonella Orlando² 

[1] Laboratory of Nutritional Biochemistry, National Institute for Digestive Diseases I.R.C.C.S. “Saverio de Bellis”, via Turi 27, I-70013 Castellana Grotte, BA, Italy;Laboratory of Nutritional Pathophysiology, National Institute for Digestive Diseases I.R.C.C.S. “Saverio de Bellis”, via Turi 27, I-70013 Castellana Grotte, BA, Italy

关键词: Tight junction proteins;    Polyamines;    Paracellular permeability;    Lactobacillus rhamnosus GG;    Intestinal barrier function;    Gliadin;    Celiac disease;    Caco-2 cells;   

Others  :  1142095
DOI  :  10.1186/1471-2180-14-19

 received in 2013-11-22, accepted in 2014-01-27,  发布年份 2014

【 摘 要 】

Background

Celiac disease is characterized by enhanced intestinal paracellular permeability due to alterations of function and expression of tight junction (TJ) proteins including ZO-1, Claudin-1 and Occludin. Polyamines are pivotal in the control of intestinal barrier function and are also involved in the regulation of intercellular junction proteins. Different probiotic strains may inhibit gliadin-induced toxic effects and the Lactobacillus rhamnosus GG (L.GG) is effective in the prevention and treatment of gastrointestinal diseases. Aims of the study were to establish in epithelial Caco-2 cells whether i) gliadin affects paracellular permeability and polyamine profile; ii) co-administration of viable L.GG, heat-killed L.GG (L.GG-HK) or its conditioned medium (L.GG-CM) preserves the intestinal epithelial barrier integrity. Additionally, the effects of L.GG on TJ protein expression were tested in presence or absence of polyamines.

Results

Administration of gliadin (1 mg/ml) to Caco-2 cells for 6 h caused a significant alteration of paracellular permeability as demonstrated by the rapid decrease in transepithelial resistance with a concomitant zonulin release. These events were followed by a significant increase in lactulose paracellular transport and a slight lowering in ZO-1 and Occludin expression without affecting Claudin-1. Besides, the single and total polyamine content increased significantly. The co-administration of viable L.GG (10⁸ CFU/ml), L.GG-HK and L.GG-CM with gliadin significantly restored barrier function as demonstrated by transepithelial resistance, lactulose flux and zonulin release. Viable L.GG and L.GG-HK, but not L.GG-CM, led to a significant reduction in the single and total polyamine levels. Additionally, only the co-administration of viable L.GG with gliadin significantly increased ZO-1, Claudin-1 and Occludin gene expression compared to control cells. When Caco-2 cells treated with viable L.GG and gliadin were deprived in the polyamine content by α-Difluoromethylornithine, the expression of TJ protein mRNAs was not significantly different from that in controls or cells treated with gliadin alone.

Conclusions

Gliadin modifies the intestinal paracellular permeability and significantly increases the polyamine content in Caco-2 cells. Concomitant administration of L.GG is able to counteract these effects. Interestingly, the presence of cellular polyamines is necessary for this probiotic to exert its capability in restoring paracellular permeability by affecting the expression of different TJ proteins.

【 授权许可】

   
2014 Orlando et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Green PHR, Cellier C: Celiac disease. N Engl J Med 2007, 357(17):1731-1743.
• [2]Mettner J: Gluten and the gut. Minn Med 2012, 95(12):14-18.
• [3]Dieterich W, Esslinger B, Schuppan D: Pathomechanisms in celiac disease. Int Arch Allergy Immunol 2003, 132(2):98-108.
• [4]Heyman M, Abed J, Lebreton C, Cerf-Bensussan N: Intestinal permeability in celiac disease: insight into mechanisms and relevance to pathogenesis. Gut 2012, 61(9):1355-1364.
• [5]Liu Y, Nusrat A, Schnell FJ, Reaves TA, Walsh S, Pochet M, Parkos CA: Human junction adhesion molecule regulates tight junction resealing in epithelia. J Cell Sci 2000, 113(Pt 13):2363-2374.
• [6]Assimakopoulos SF, Papageorgiou I, Charonis A: Enterocytes’ tight junctions: from molecules to diseases. World J Gastrointest Pathophysiol 2011, 2(6):123-137.
• [7]Al-Sadi R, Khatib K, Guo S, Ye D, Youssef M, Ma T: Occludin regulates macromolecule flux across the intestinal epithelial tight junction barrier. Am J Physiol Gastrointest Liver Physiol 2011, 300(6):G1054-G1064.
• [8]Zeissig S, Bürgel N, Günzel D, Richter J, Mankertz J, Wahnschaffe U, Kroesen AJ, Zeitz M, Fromm M, Schulzke JD: Changes in expression and distribution of claudin 2,5, and 8 lead to discontinuous tight junctions and barrier dysfunction in active Crohn’s disease. Gut 2007, 56(1):61-72.
• [9]McCall IC, Betanzos A, Weber DA, Nava P, Miller GW, Parkos CA: Effects of phenol on barrier function of a human intestinal epithelial cell line correlate with altered tight junction protein localization. Toxicol Appl Pharmacol 2009, 241:61-70.
• [10]Assimakopoulos SF, Tsamandas AC, Tsiaoussis GI, Karatza E, Triantos C, Vagianos CE, Spiliopoulou I, Kaltezioti V, Charonis A, Nikolopoulou VN, Scopa CD, Thomopoulos KC: Altered intestinal tight junctions’ expression in patients with liver cirrhosis: a pathogenetic mechanism of intestinal hyperpermeability. Eur J Clin Invest 2012, 42(4):439-446.
• [11]Thomas T, Thomas TJ: Polyamines in cell growth and cell death: molecular mechanisms and therapeutic applications. Cell Mol Life Sci 2001, 58:244-258.
• [12]Wang JY: Polyamines regulate expression of E-cadherin and play an important role in control of intestinal epithelial barrier function. Inflammopharmacology 2005, 13(1–3):91-101.
• [13]Osman NE, Weström B, Wang Q, Persson L, Karlsson B: Spermine affects intestinal in vitro permeability to different-sized molecules in rats. Comp Biochem Physiol C Pharmacol Toxicol Endocrinol 1998, 120(2):211-216.
• [14]Auricchio S, De Ritis G, De Vincenzi M, Gentile V, Maiuri L, Mancini E, Porta R, Raia V: Amines protect in vitro the celiac small intestine from the damaging activity of gliadin peptides. Gastroenterology 1990, 99(6):1668-1674.
• [15]Madsen K, Cornish A, Soper P, McKaigney C, Jijon H, Yachimec C, Doyle J, Jewell L, De Simone C: Probiotic bacteria enhance murine and human intestinal epithelial barrier function. Gastroenterology 2001, 121:580-591.
• [16]Gupta P, Andrew H, Kirschner BS, Guandalini S: Is lactobacillus GG helpful in children with Crohn’s disease? Results of a preliminary, open-label study. J Pediatr Gastroenterol Nutr 2000, 31:453-457.
• [17]Lindfors K, Blomqvist T, Juuti-Uusitalo K, Stenman S, Venäläinen J, Mäki M, Kaukinen K: Live probiotic Bifidobacterium lactis bacteria inhibit the toxic effects induced by wheat gliadin in epithelial cell culture. Clin Exp Immunol 2008, 152:552-558.
• [18]Marteau PR, de Vrese M, Cellier CJ, Schrezenmeir J: Protection from gastrointestinal diseases with the use of probiotics. Am J Clin Nutr 2001, 73(2 Suppl):430S-436S.
• [19]Orlando A, Messa C, Linsalata M, Cavallini A, Russo F: Effects of Lactobacillus rhamnosus GG on proliferation and polyamine metabolism in HGC-27 human gastric and DLD-1 colonic cancer cell lines. Immunopharmacol Immunotoxicol 2009, 31(1):108-116.
• [20]Orlando A, Refolo MG, Messa C, Amati L, Lavermicocca P, Guerra V, Russo F: Antiproliferative and proapoptotic effects of viable or heat-killed Lactobacillus paracasei IMPC2.1 and Lactobacillus rhamnosus GG in HGC-27 gastric and DLD-1 colon cell lines. Nutr Cancer 2012, 64(7):1103-1111.
• [21]Vachon PH, Beaulieu JF: Transient mosaic patterns of morphological and functional differentiation in Caco-2 cell line. Gastroenterology 1991, 103:414-423.
• [22]Drago S, El Asmar R, Di Pierro M, Grazia Clemente M, Tripathi A, Sapone A, Thakar M, Iacono G, Carroccio A, D’Agate C, Not T, Zampini L, Catassi C, Fasano A: Gliadin, zonulin and gut permeability: effects on celiac and non-celiac intestinal mucosa and intestinal cell lines. Scand J Gastroenterol 2006, 41(4):408-419.
• [23]El Asmar R, Panigrahi P, Bamford P, Berti I, Not T, Coppa GV, Catassi C, Fasano A: Host-dependent zonulin secretion causes the impairment of the small intestine barrier function after bacterial exposure. Gastroenterology 2002, 123(5):1607-1615.
• [24]Linsalata M, Russo F, Notarnicola M, Berloco P, Di Leo A: Polyamine profile in human gastric mucosa infected by helicobacter pylori. Ital J Gastroenterol Hepatol 1998, 30(5):484-489.
• [25]Guo X, Rao JN, Liu L, Zou T, Keledjian KM, Boneva D, Marasa BS, Wang JY: Polyamines are necessary for synthesis and stability of occludin protein in intestinal epithelial cells. Am J Physiol Gastrointest Liver Physiol 2005, 288(6):G1159-G1169.
• [26]Linsalata M, Notarnicola M, Tutino V, Bifulco M, Santoro A, Laezza C, Messa C, Orlando A, Caruso MG: Effects of anandamide on polyamine levels and cell growth in human colon cancer cells. Anticancer Res 2010, 30(7):2583-2589.
• [27]Di Cagno R, De Angelis M, Auricchio S, Greco L, Clarke C, De Vincenzi M, Giovannini C, D’Archivio M, Landolfo F, Parrilli G, Minervini F, Arendt E, Gobbetti M: Sourdough bread made from wheat and nontoxic flours and started with selected lactobacilli is tolerated in celiac sprue patients. Appl Environ Microbiol 2004, 70(2):1088-1096.
• [28]Koch S, Nusrat A: Dynamic regulation of epithelial cell fate and barrier function by intercellular junctions. Ann N Y Acad Sci 2009, 1165:220-227.
• [29]Sander GR, Cummins AG, Henshall T, Powell BC: Rapid disruption of intestinal barrier function by gliadin involves altered expression of apical junctional proteins. FEBS Lett 2005, 579(21):4851-4855.
• [30]Rivabene R, Mancini E, De Vincenzi M: In vitro cytotoxic effect of wheat gliadin-derived peptides on the Caco-2 intestinal cell line is associated with intracellular oxidative imbalance: implications for coeliac disease. Biochim Biophys Acta (BBA) - Mol Basis Dis 1999, 1453(1):152-160.
• [31]Elgavish A, Wallace RW, Pillion DJ, Meezan E: Polyamines stimulate D-glucose transport in isolated renal brush-border membrane vesicles. Biochimica et biophysica acta 1984, 777(1):1-8.
• [32]Matysiak-Budnik T, Moura IC, Arcos-Fajardo M, Lebreton C, Ménard S, Candalh C, Ben-Khalifa K, Dugave C, Tamouza H, van Niel G, Bouhnik Y, Lamarque D, Chaussade S, Malamut G, Cellier C, Cerf-Bensussan N, Monteiro RC, Heyman M: Secretory IgA mediates retrotranscytosis of intact gliadin peptides via the transferrin receptor in celiac disease. J Exp Med 2008, 205:143-154.
• [33]Colyer J, Kumar PJ, Waldron NM, Clark ML, Farthing MJ: Farthing Gliadin binding to rat and human enterocytes. Clin Sci 1987, 12:593-598.
• [34]Peulen O, Deloyer P, Deville C, Dandrifosse G: Polyamines in Gut inflammation and allergy. Curr Med Chem Anti-Inflamm Anti-Allergy Agents 2004, 3(1):1-8.
• [35]Ahrné S, Nobaek S, Jeppsson B, Adlerberth I, Wold AE, Molin G: The normal lactobacillus flora of healthy human rectal and oral mucosa. J Appl Microbiol 1998, 85(1):88-94.
• [36]Ahrne S, Hagslatt ML: Effect of lactobacilli on paracellular permeability in the gut. Nutr 2011, 3(1):104-117.
• [37]Seth A, Yan F, Polk DB, Rao RK: Probiotics ameliorate the hydrogen peroxide-induced epithelial barrier disruption by a PKC- and MAP kinase-dependent mechanism. Am J Physiol Gastrointest Liver Physiol 2008, 294(4):G1060-G1069.
• [38]Linsalata M, Cavallini A, Messa C, Orlando A, Refolo MG, Russo F: Lactobacillus rhamnosus GG influences polyamine metabolism in HGC-27 gastric cancer cell line: a strategy toward nutritional approach to chemoprevention of gastric cancer. Curr Pharm Des 2010, 16(7):847-853.
• [39]Di Cagno R, De Angelis M, Lavermicocca P, De Vincenzi M, Giovannini C, Faccia M, Gobbetti M: Proteolysis by sourdough lactic acid bacteria: effects on wheat flour protein fractions and gliadin peptides involved in human cereal intolerance. Appl Environ Microbiol 2002, 68:623-633.
• [40]De Angelis M, Rizzello CG, Fasano A, Clemente MG, De Simone C, Silano M, De Vincenzi M, Losito I, Gobbetti M: VSL3# probiotic preparation has the capacity to hydrolyse gliadin polypeptides responsible for celiac sprue. Biochim Biophys Acta 2005, 1762:80-93.
• [41]Lee YK, Ho PS, Low CS, Arvilommi H, Salminen S: Permanent colonization by lactobacillus casei is hindered by the low rate of cell division in mouse gut. Appl Environ Microbiol 2004, 70(2):670-674.
• [42]Valeur N, Engel P, Carbajal N, Connolly E, Ladefoged K: Colonization and immunomodulation by lactobacillus reuteri ATCC 55730 in the human gastrointestinal tract. Appl Environ Microbiol 2004, 70(2):1176-1181.
• [43]Claes IJ, Schoofs G, Regulski K, Courtin P, Chapot-Chartier MP, Rolain T, Hols P, von Ossowski I, Reunanen J, de Vos WM, Palva A, Vanderleyden J, De Keersmaecker SC, Lebeer S: Genetic and biochemical characterization of the cell wall hydrolase activity of the major secreted protein of Lactobacillus rhamnosus GG. PLoS One 2012, 7(2):e31588.
• [44]Klingberg TD, Pedersen MH, Cencic A, Budde BB: Application of measurement of transepithelial electrical resistance of intestinal epithelial cell monolayers to evaluate probiotic activity. Appl Environ Microbiol 2005, 71:7528-7530.
• [45]Karczewski J, Troost FJ, Konings I, Dekker J, Kleerebezem M, Brummer RJ, Wells JM: Regulation of human epithelial tight junction proteins by Lactobacillus plantarum in vivo and protective effects on the epithelial barrier. Am J Physiol Gastrointest Liver Physiol 2010, 298:G851-G859.
• [46]Anderson RC, Cookson AL, McNabb WC, Park Z, McCann MJ, Kelly WJ, Roy NC: Lactobacillus plantarum MB452 enhances the function of the intestinal barrier by increasing the expression levels of genes involved in tight junction formation. BMC Microbiol 2010, 10:316. BioMed Central Full Text
• [47]Tabor CW, Tabor H: Polyamines. Annu Rev Biochem 1984, 53:749-790.
• [48]Verdu EF, Huang X, Natividad J, Lu J, Blennerhassett PA, David CS, McKay DM, Murray JA: Gliadin-dependent neuromuscular and epithelial secretory responses in gluten-sensitive HLA-DQ8 transgenic mice. Am J Physiol Gastrointest Liver Physiol 2008, 294(1):G217-G225.