Gliadin Intake Causes Disruption of the Intestinal Barrier and an Increase in Germ Cell Apoptosis in A Caenorhabditis Elegans Model
Abstract
:1. Introduction
2. Materials and Methods
2.1. C. Elegans Strains and Gliadin Treatment
2.2. Live Image Observation of Fluorescence-Tagged Transgenic Worms
2.3. Treatment with Synthetic Gliadin Peptides and Wheat Gluten Hydrolysate (WGH)
2.4. Reactive Oxygen Species (ROS) Measurements
2.5. Phalloidin Staining
2.6. Intestinal Barrier Function Assay
2.7. N-Acetyl-L-Cysteine (NAC) Treatment
2.8. Measurement of Brood Size
2.9. Germ Cell Apoptosis Assay
2.10. RNA Interference (RNAi) Assays
2.11. Immunofluorescence Analysis
2.12. Cell Culture and Cell Viability Assays
2.13. Intracellular ROS Accumulation Measurements
2.14. Statistical Analysis
3. Results
3.1. Gliadin Intake Induces GST-4, CYP-35 and ROS Production in Adult-Stage C. elegans
3.2. ROS Production Is Induced by the Intake of Synthetic Gliadin Peptides but not Wheat Gluten Hydrolysate (WGH) in Adult-Stage C. elegans
3.3. Intestinal Integrity is Disrupted by Gliadin Intake
3.4. Effects of Gliadin and Synthetic Gliadin Peptides on Intestinal F-Actin Formation Are Suppressed by Antioxidant Treatment
3.5. Gliadin Intake Increases Germ Cell Apoptosis
3.6. Effects of Gliadin and Synthetic Gliadin Peptides on Germ Cell Apoptosis are Suppressed by Antioxidant Treatment
3.7. Effect of Antioxidants on Gliadin-Induced ROS and Germ Cell Apoptosis in An Oxidative Stress-Sensitive Mutant
4. Discussion
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Volta, U.; De Giorgio, R. New understanding of gluten sensitivity. Nat. Rev. Gastroenterol. Hepatol. 2012, 9, 295–299. [Google Scholar] [CrossRef] [PubMed]
- Sapone, A.; Bai, J.C.; Ciacci, C.; Dolinsek, J.; Green, P.H.; Hadjivassiliou, M.; Kaukinen, K.; Rostami, K.; Sanders, D.S.; Schumann, M.; et al. Spectrum of gluten-related disorders: Consensus on new nomenclature and classification. BMC Med. 2012, 7, 10–13. [Google Scholar] [CrossRef] [PubMed]
- Volta, U.; Caio, G.; Tovoli, F.; De Giorgio, R. Non-celiac gluten sensitivity: Questions still to be answered despite increasing awareness. Cell. Mol. Immunol. 2013, 10, 383–392. [Google Scholar] [CrossRef] [PubMed]
- Leonard, M.M.; Vasagar, B. US perspective on gluten-related diseases. Clin. Exp. Gastroenterol. 2014, 7, 25–37. [Google Scholar] [CrossRef] [Green Version]
- Nanayakkara, M.; Lania, G.; Maglio, M.; Discepolo, V.; Sarno, M.; Gaito, A.; Troncone, R.; Auricchio, S.; Auricchio, R.; Barone, M.V. An undigested gliadin peptide activates innate immunity and proliferative signaling in enterocytes: The role in celiac disease. Am. J. Clin. Nutr. 2013, 4, 1123–1135. [Google Scholar] [CrossRef]
- Nikulina, M.; Habich, C.; Flohé, S.B.; Scott, F.W.; Kolb, H. Wheat gluten causes dendritic cell maturation and chemokine secretion. J. Immunol. 2004, 3, 1925–1933. [Google Scholar] [CrossRef]
- Fasano, A. Zonulin and its regulation of intestinal barrier function: The biological door to inflammation, autoimmunity, and cancer. Physiol. Rev. 2011, 1, 151–175. [Google Scholar] [CrossRef]
- Maiuri, L.; Troncone, R.; Mayer, M.; Coletta, S.; Picarell, i.A.; De Vincenzi, M.; Pavone, V.; Auricchio, S. In vitro activities of A-gliadin-related synthetic peptides: Damaging effect on the atrophic coeliac mucosa and activation of mucosal immune response in the treated coeliac mucosa. Scand. J. Gastroenterol. 1996, 3, 247–253. [Google Scholar] [CrossRef]
- Vilasi, S.; Sirangelo, I.; Irace, G.; Caputo, I.; Barone, M.V.; Esposito, C.; Ragone, R. Interaction of ‘toxic’ and ‘immunogenic’ A-gliadin peptides with a membrane-mimetic environment. J. Mol. Recognit. 2010, 3, 322–328. [Google Scholar] [CrossRef]
- Camarca, A.; Anderson, R.P.; Mamone, G.; Fierro, O.; Facchiano, A.; Costantini, S.; Zanzi, D.; Sidney, J.; Auricchio, S.; Sette, A.; et al. Intestinal T cell responses to gluten peptides are largely heterogeneous: Implications for a peptide-based therapy in celiac disease. J. Immunol. 2009, 182, 4158–4166. [Google Scholar] [CrossRef]
- Shan, L.; Molberg, Ø.; Parrot, I.; Hausch, F.; Filiz, F.; Gray, G.M.; Sollid, L.M.; Khosla, C. Structural basis for gluten intolerance in celiac sprue. Science 2002, 297, 2275–2279. [Google Scholar] [CrossRef] [PubMed]
- Lammers, K.M.; Lu, R.; Brownley, J.; Lu, B.; Gerard, C.; Thomas, K.; Rallabhandi, P.; Shea-Donohue, T.; Tamiz, A.; Alkan, S.; et al. Gliadin induces an increase in intestinal permeability and zonulin release by binding to the chemokine receptor CXCR3. Gastroenterology 2008, 135, 194–204. [Google Scholar] [CrossRef] [PubMed]
- Lammers, K.M.; Khandelwal, S.; Kryszak, D.; Casolaro, V.; Fasano, A. PBMC from celiac patients but not healthy controls produce interleukin-8 in response to gliadin that is CXCR3-dependent. Gastroenterology 2009, 136, A472. [Google Scholar] [CrossRef]
- Hollon, J.; Puppa, E.L.; Greenwald, B.; Goldberg, E.; Guerrerio, A.; Fasano, A. Effect of gliadin on permeability of intestinal biopsy explants from celiac disease patients and patients with non-celiac gluten sensitivity. Nutrients 2015, 3, 1565–1576. [Google Scholar] [CrossRef]
- Lim, S.D.; Min, H.; Youn, E.; Kawasaki, I.; Shim, Y.H. Gliadin intake induces oxidative-stress response in Caenorhabditis elegans. Biochem. Biophys. Res. Commun. 2018, 3, 2139–2145. [Google Scholar] [CrossRef]
- He, F.; Zuo, L. Redox roles of reactive oxygen species in cardiovascular diseases. Int. J. Mol. Sci. 2015, 11, 27770–27780. [Google Scholar] [CrossRef]
- Zuo, L.; Zhou, T.; Pannell, B.K.; Ziegler, A.C.; Best, T.M. Biological and physiological role of reactive oxygen species--the good, the bad and the ugly. Acta Physiol. (Oxf.) 2015, 3, 329–348. [Google Scholar] [CrossRef]
- Hodgkin, J.; Plasterk, R.H.; Waterston, R.H. The nematode Caenorhabditis elegans and its genome. Science 1995, 270, 410–414. [Google Scholar] [CrossRef]
- Calvo, D.R.; Martorell, P.; Genoves, S.; Luis, G. Development of novel functional ingredients: Need for testing systems and solutions with Caenorhabditis elegans. Trends. Food Sci. Technol. 2016, 54, 197–203. [Google Scholar] [CrossRef]
- Gruber, J.; Ng, L.F.; Poovathingal, S.K.; Halliwell, B. Deceptively simple but simply deceptive—Caenorhabditis elegans lifespan studies: Considerations for aging and antioxidant effects. FEBS Lett. 2009, 21, 3377–3387. [Google Scholar] [CrossRef]
- Brenner, S. The genetics of Caenorhabditis elegans. Genetics 1974, 77, 71–94. [Google Scholar] [PubMed]
- Zhang, W.; Lv, T.; Li, M.; Wu, Q.; Yang, L.; Liu, H.; Sun, D.; Zhuang, Z.; Wang, D. Beneficial effects of wheat gluten hydrolysate to extend lifespan and induce stress resistance in nematode Caenorhabditis elegans. PLoS ONE 2013, 8, e74553. [Google Scholar] [CrossRef] [PubMed]
- Ruan, Q.; Qiao, Y.; Zhao, Y.; Xu, Y.; Wang, M.; Duan, J.; Wang, D. Beneficial effects of Glycyrrhizae radix extract in preventing oxidative damage and extending the lifespan of Caenorhabditis elegans. J. Ethnopharmacol. 2016, 177, 101–110. [Google Scholar] [CrossRef] [PubMed]
- LeBel, C.P.; Ischiropoulos, H.; Bondy, S.C. Evaluation of the probe 2′,7′-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress. Chem. Res. Toxicol. 1992, 5, 227–231. [Google Scholar] [CrossRef]
- Strome, S. Fluorescence visualization of the distribution of microfilaments in gonads and early embryos of the nematode Caenorhabditis elegans. J. Cell Biol. 1986, 103, 2241–2252. [Google Scholar] [CrossRef]
- Gelino, S.; Chang, J.T.; Kumsta, C.; She, X.; Davis, A.; Nguyen, C.; Panowski, S.; Hansen, M. Intestinal autophagy improves health span and longevity in C. elegans during dietary restriction. PLoS Genet. 2016, 12, e1006135. [Google Scholar] [CrossRef]
- Navarro, R.E.; Shim, E.Y.; Kohara, Y.; Singson, A.; Blackwell, T.K. cgh-1, a conserved predicted RNA helicase required for gametogenesis and protection from physiological germline apoptosis in C. elegans. Development 2001, 128, 3221–3232. [Google Scholar]
- Min, H.; Shim, Y.H.; Kawasaki, I. Loss of PGL-1 and PGL-3, members of a family of constitutive germ-granule components, promotes germline apoptosis in C. elegans. J. Cell Sci. 2016, 129, 341–353. [Google Scholar] [CrossRef]
- Leiers, B.; Kampkötter, A.; Grevelding, C.G.; Link, C.D.; Johnson, T.E.; Henkle-Dührsen, K. A stress-responsive glutathione S-transferase confers resistance to oxidative stress in Caenorhabditis elegans. Free Radic. Biol. Med. 2003, 34, 1405–1415. [Google Scholar] [CrossRef]
- Ayyadevara, S.; Dandapat, A.; Singh, S.P.; Siegel, E.R.; Shmookler Reis, R.J.; Zimniak, L.; Zimniak, P. Life span and stress resistance of Caenorhabditis elegans are differentially affected by glutathione transferases metabolizing 4-hydroxynon-2-enal. Mech. Ageing Dev. 2007, 128, 196–205. [Google Scholar] [CrossRef]
- Dottermusch, M.; Lakner, T.; Peyman, Y.; Klein, M.; Walz, G.; Neumann-Haefelin, E. Cell cycle controls stress response and longevity in C. Elegans. Aging 2016, 8, 2100–2115. [Google Scholar] [CrossRef] [PubMed]
- Picarelli, A.; Libanori, V.; De Nitto, D.; Saponara, A.; Di Tola, M.; Donato, G. Organ culture system as a means to detect celiac disease. Ann. Clin. Lab. Sci. 2010, 20, 85–87. [Google Scholar]
- Emerit, J.; Pelletier, S.; Tosoni-Verlignue, D.; Mollet, M. Phase II trial of copper zinc superoxide dismutase (CuZnSOD) in treatment of Crohn’s disease. Free Radic. Biol. Med. 1989, 7, 145–149. [Google Scholar] [CrossRef]
- Clark, D.A.; Fornabaio, D.M.; McNeill, H.; Mullane, K.M.; Caravella, S.J.; Miller, M.J. Contribution of oxygen-derived free radicals to experimental necrotizing enterocolitis. Am. J. Pathol. 1988, 130, 537–542. [Google Scholar]
- Rao, R. Oxidative stress-induced disruption of epithelial and endothelial tight junctions. Front. Biosci. 2008, 13, 7210–7226. [Google Scholar] [CrossRef]
- Gangwar, R.; Meena, A.S.; Shukla, P.K.; Nagaraja, A.S.; Dorniak, P.L.; Pallikuth, S.; Waters, C.M.; Sood, A.; Rao, R. Calcium-mediated oxidative stress: A common mechanism in tight junction disruption by different types of cellular stress. Biochem. J. 2017, 474, 731–749. [Google Scholar] [CrossRef]
- Bossinger, O.; Fukushige, T.; Claeys, M.; Borgonie, G.; McGhee, J.D. The apical disposition of the Caenorhabditis elegans intestinal terminal web is maintained by LET-413. Dev. Biol. 2004, 268, 448–456. [Google Scholar] [CrossRef]
- Shukla, P.K.; Gangwar, R.; Manda, B.; Meena, A.S.; Yadav, N.; Szabo, E.; Balogh, A.; Lee, S.C.; Tigyi, G.; Rao, R. Rapid disruption of intestinal epithelial tight junction and barrier dysfunction by ionizingradiation in mousecolon invivo: Protection by N-acetyl-l-cysteine. Am. J. Physiol. Gastrointest. Liver Physiol. 2016, 310, G705–G715. [Google Scholar] [CrossRef]
- Balklava, Z.; Rathnakumar, N.D.; Vashist, S.; Schweinsberg, P.J.; Grant, B.D. Linking gene expression in the intestine to production of gametes through the phosphate transporter PITR-1 in Caenorhabditis elegans. Genetics 2016, 204, 153–162. [Google Scholar] [CrossRef]
- O’Rourke, E.J.; Soukas, A.A.; Carr, C.E.; Ruvkun, G. C. elegans major fats are stored in vesicles distinct from lysosome-related organelles. Cell Metab. 2009, 10, 430–435. [Google Scholar] [CrossRef]
- Steinbaugh, M.J.; Narasimhan, S.D.; Robida-Stubbs, S.; Moronetti Mazzeo, L.E.; Dreyfuss, J.M.; Hourihan, J.M.; Raghavan, P.; Operaña, T.N.; Esmaillie, R.; Blackwell, T.K. Lipid-mediated regulation of SKN-1/Nrf in response to germ cell absence. eLife 2015, 4, e07836. [Google Scholar] [CrossRef] [PubMed]
- Derry, W.B.; Putzke, A.P.; Rothman, J.H. Caenorhabditis elegans p53: Role in apoptosis, meiosis, and stress resistance. Science 2001, 294, 591–595. [Google Scholar] [CrossRef] [PubMed]
- Levi-Ferber, M.; Salzberg, Y.; Safra, M.; Haviv-Chesner, A.; Bülow, H.E.; Henis-Korenblit, S. It’s all in your mind: Determining germ cell fate by neuronal IRE-1 in C. elegans. PLoS Genet. 2014, 10, e1004747. [Google Scholar] [CrossRef] [PubMed]
- Hofmann, E.R.; Milstein, S.; Boulton, S.J.; Ye, M.; Hofmann, J.J.; Stergiou, L.; Gartner, A.; Vidal, M.; Hengartner, M.O. Caenorhabditis elegans HUS-1 is a DNA damage checkpoint protein required for genome stabilityand EGL-1 mediated apoptosis. Curr. Biol. 2002, 12, 1908–1918. [Google Scholar] [CrossRef]
- Peyssonnaux, C.; Eychene, A. The Raf/MEK/ERK pathway: New concepts of activation. Biol. Cell. 2001, 93, 53–62. [Google Scholar] [CrossRef]
- Wang, J.; Du, H.; Nie, Y.; Wang, Y.; Dai, H.; Wang, M.; Wang, D.; Xu, A. Mitochondria and MAPK cascades modulate endosulfan induced germline apoptosis in Caenorhabditis elegans. Toxicol. Res. (Camb.) 2017, 6, 412–419. [Google Scholar] [CrossRef]
- Sijen, T.; Fleenor, J.; Simmer, F.; Thijssen, K.L.; Parrish, S.; Timmons, L.; Plasterk, R.H.; Fire, A. On the role of RNA amplification in dsRNA-triggered gene silencing. Cell 2001, 107, 465–476. [Google Scholar] [CrossRef]
- Tijsterman, M.; Okihara, K.L.; Thijssen, K.; Plasterk, R.H. PPW-1, a PAZ/PIWI protein required for efficient germline RNAi, is defective in a natural isolate of C. elegans. Curr. Biol. 2002, 12, 1535–1540. [Google Scholar] [CrossRef]
- Ishii, N.; Fujii, M.; Hartman, P.S.; Tsuda, M.; Yasuda, K.; Senoo-Matsuda, N.; Yanase, S.; Ayusawa, D.; Suzuki, K. A mutation in succinate dehydrogenase cytochrome b causes oxidative stress and ageing in nematodes. Nature 1998, 394, 694–697. [Google Scholar] [CrossRef]
- Corsi, A.K.; Wightman, B.; Chalfie, M. A transparent window into biology: A primer on Caenorhabditis elegans. Genetics 2015, 200, 387–407. [Google Scholar] [CrossRef]
- The C. elegans Sequencing Consortium. Genome sequence of the nematode C. elegans: A platform for investigating biology. Science 1998, 282, 2012–2018. [Google Scholar] [CrossRef] [PubMed]
- Lai, C.H.; Chou, C.Y.; Ch’ang, L.Y.; Liu, C.S.; Lin, W. Identification of novel human genes evolutionarily conserved in Caenorhabditis elegans by comparative proteomics. Genome Res. 2000, 10, 703–713. [Google Scholar] [CrossRef] [PubMed]
- Kaletta, T.; Hengartner, M.O. Finding function in novel targets: C. elegans as a model organism. Nat. Rev. Drug. Discov. 2006, 5, 387–398. [Google Scholar] [CrossRef] [PubMed]
- Sander, G.R.; Cummins, A.G.; Henshall, T.; Powell, B.C. Rapid disruption of intestinal barrier function by gliadin involves altered expression of apicaljunctional proteins. FEBS Lett. 2005, 579, 4851–4855. [Google Scholar] [CrossRef] [PubMed]
- Zhang, L.; Andersen, D.; Roager, H.M.; Bahl, M.I.; Hansen, C.H.; Danneskiold-Samsøe, N.B.; Kristiansen, K.; Radulescu, I.D.; Sina, C.; Frandsen, H.L.; et al. Effects of gliadin consumption on the intestinal microbiota and metabolic homeostasis in mice fed a high-fat diet. Sci. Rep. 2017, 7, 44613. [Google Scholar] [CrossRef] [PubMed]
- Cabello-Verrugio, C.; Simon, F.; Trollet, C.; Santibañez, J.F. Oxidative stress in disease and aging: Mechanisms and therapies 2016. Oxid. Med. Cell. Longev. 2017, 2017, 4310469. [Google Scholar] [CrossRef] [PubMed]
- Bergamo, P.; Maurano, F.; Mazzarella, G.; Iaquinto, G.; Vocca, I.; Rivelli, A.R.; De Falco, E.; Gianfrani, C.; Rossi, M. Immunological evaluation of the alcohol-soluble protein fraction from gluten free grains in relation to celiac disease. Mol. Nutr. Food Res. 2011, 55, 1266–1270. [Google Scholar] [CrossRef]
- Dolfini, E.; Elli, L.; Dasdia, T.; Bufardeci, B.; Colleoni, M.P.; Costa, B.; Floriani, I.; Falini, M.L.; Guerrieri, N.; Forlani, F.; et al. In vitro cytotoxic effect of bread wheat gliadin on the LoVo human adenocarcinoma cell line. Toxicol. In Vitro 2002, 16, 331–337. [Google Scholar] [CrossRef]
- Maiuri, L.; Ciacci, C.; Ricciardelli, I.; Vacca, L.; Raia, V.; Auricchio, S.; Picard, J.; Osman, M.; Quaratino, S.; Londei, M. Association between innate response to gliadin and activation of pathogenic T cells in coeliac disease. Lancet 2003, 362, 30–37. [Google Scholar] [CrossRef]
- Stojiljković, V.; Todorović, A.; Pejić, S.; Kasapović, J.; Saicić, Z.S.; Radlović, N.; Pajović, S.B. Antioxidant status and lipid peroxidation in small intestinal mucosa of children with celiac disease. Clin. Biochem. 2009, 42, 1431–1437. [Google Scholar] [CrossRef]
- Rowicka, G.; Czaja-Bulsa, G.; Chełchowska, M.; Riahi, A.; Strucińska, M.; Weker, H.; Ambroszkiewicz, J. Oxidative and antioxidative status of children with celiac disease treated with a gluten free-diet. Oxid. Med. Cell. Longev. 2018, 2018, 1324820. [Google Scholar] [CrossRef] [PubMed]
- Salinas, L.S.; Maldonado, E.; Navarro, R.E. Stress-induced germ cell apoptosis by a p53 independent pathway in Caenorhabditis elegans. Cell Death Differ. 2006, 13, 2129–2139. [Google Scholar] [CrossRef] [PubMed]
- McCubrey, J.A.; Steelman, L.S.; Abrams, S.L.; Lee, J.T.; Chang, F.; Bertrand, F.E.; Navolanic, P.M.; Terrian, D.M.; Franklin, R.A.; D’Assoro, A.B.; et al. Roles of the RAF/MEK/ERK and PI3K/PTEN/AKT pathways in malignant transformation and drug resistance. Adv. Enzyme Regul. 2006, 46, 249–279. [Google Scholar] [CrossRef] [PubMed]
- Ruffels, J.; Griffin, M.; Dickenson, J.M. Activation of ERK1/2, JNK and PKB by hydrogen peroxide in human SH-SY5Y neuroblastomacells: Role of ERK1/2 in H2O2-induced cell death. Eur. J. Pharmacol. 2004, 483, 163–173. [Google Scholar] [CrossRef]
- Riddle, D.L.; Albert, P.S. Patterning the nervous system. In C. elegans II, 2nd ed.; Riddle, D.L., Blumenthal, T., Meyer, B.J., Priess, J.R., Eds.; Cold Spring Harbor Laboratory Press: New York, NY, USA, 1997; pp. 543–547. ISBN 0-87969-488-2. [Google Scholar]
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Min, H.; Kim, J.-S.; Ahn, J.; Shim, Y.-H. Gliadin Intake Causes Disruption of the Intestinal Barrier and an Increase in Germ Cell Apoptosis in A Caenorhabditis Elegans Model. Nutrients 2019, 11, 2587. https://doi.org/10.3390/nu11112587
Min H, Kim J-S, Ahn J, Shim Y-H. Gliadin Intake Causes Disruption of the Intestinal Barrier and an Increase in Germ Cell Apoptosis in A Caenorhabditis Elegans Model. Nutrients. 2019; 11(11):2587. https://doi.org/10.3390/nu11112587
Chicago/Turabian StyleMin, Hyemin, Ji-Sun Kim, Jiyun Ahn, and Yhong-Hee Shim. 2019. "Gliadin Intake Causes Disruption of the Intestinal Barrier and an Increase in Germ Cell Apoptosis in A Caenorhabditis Elegans Model" Nutrients 11, no. 11: 2587. https://doi.org/10.3390/nu11112587