Abstract
Gut hormones produced by entero-endocrine cells (EEC) located throughout the gastrointestinal tract play a major role in the regulation of glucose and energy homeostasis. Angiopoietin-like 4 (ANGPTL4, also referred to as fasting induced adipose factor) is a secreted factor involved in regulation of lipid homeostasis and has been proposed as circulating mediator between the gut microbiota and fat storage in adipose tissue, although discordant data exist. Currently, little is known about the site and regulation of ANGPTL4 production in the intestine. Here, we show using immunohistochemistry and immunofluorescence that cells positive for ANGPTL4 are scattered along the epithelial layer in the human small and large intestine. ANGPTL4-positive cells exhibit typical features of EEC characterized by large ANGPTL4-positive secretory granules directed towards the basolateral side. In support, extensive overlap was observed between ANGPTL4-positive cells and cells positive for the entero-endocrine marker chromogranin A. Higher resolution images revealed that ANGPTL4 and chromogranin A are partially present in distinct intracellular vesicles. Using entero-endocrine HuTu-80 cells, ANGPTL4 secretion was shown to be induced by short chain fatty acids and reduced by bile acids. Finally, levels of ANGPTL4 in human plasma were significantly decreased following meal consumption. In conclusion, ANGPTL4 is produced by EEC in human intestine and expression may be regulated by short chain fatty acids and bile acids.
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Alex S, Lange K, Amolo T, Grinstead JS, Haakonsson AK, Szalowska E, Koppen A, Mudde K, Haenen D, Al-Lahham S, Roelofsen H, Houtman R, van der Burg B, Mandrup S, Bonvin AM, Kalkhoven E, Muller M, Hooiveld GJ, Kersten S (2013) Short-chain fatty acids stimulate angiopoietin-like 4 synthesis in human colon adenocarcinoma cells by activating peroxisome proliferator-activated receptor gamma. Mol Cell Biol 33:1303–1316
Aronsson L, Huang Y, Parini P, Korach-Andre M, Hakansson J, Gustafsson JA, Pettersson S, Arulampalam V, Rafter J (2010) Decreased fat storage by Lactobacillus paracasei is associated with increased levels of Angiopoietin-like 4 protein (ANGPTL4). PLoS One 5:e13087
Backhed F, Ding H, Wang T, Hooper LV, Koh GY, Nagy A, Semenkovich CF, Gordon JI (2004) The gut microbiota as an environmental factor that regulates fat storage. Proc Natl Acad Sci USA 101:15718–15723
Bogunovic M, Dave SH, Tilstra JS, Chang DT, Harpaz N, Xiong H, Mayer LF, Plevy SE (2007) Enteroendocrine cells express functional Toll-like receptors. Am J Physiol Gastrointest Liver Physiol 292:G1770–G1783
Bradley WD, Zwingelstein C, Rondinone CM (2011) The emerging role of the intestine in metabolic diseases. Arch Physiol Biochem 117:165–176
Carulli N, Ponz de Leon M, Podda M, Zuin M, Strata A, Frigerio G, Digrisolo A (1981) Chenodeoxycholic acid and ursodeoxycholic acid effects in endogenous hypertriglyceridemias. A controlled double-blind trial. J Clin Pharmacol 21:436–442
Drucker DJ (2007) The role of gut hormones in glucose homeostasis. J Clin Invest 117:24–32
El Aidy S, Merrifield CA, Derrien M, van Baarlen P, Hooiveld G, Levenez F, Dore J, Dekker J, Holmes E, Claus SP, Reijngoud DJ, Kleerebezem M (2013) The gut microbiota elicits a profound metabolic reorientation in the mouse jejunal mucosa during conventionalisation. Gut 62:1306–1314
Fleissner CK, Huebel N, Abd El-Bary MM, Loh G, Klaus S, Blaut M (2010) Absence of intestinal microbiota does not protect mice from diet-induced obesity. Br J Nutr 104:919–929
Georgiadi A, Lichtenstein L, Degenhardt T, Boekschoten MV, van Bilsen M, Desvergne B, Muller M, Kersten S (2010) Induction of cardiac Angptl4 by dietary fatty acids is mediated by peroxisome proliferator-activated receptor beta/delta and protects against fatty acid-induced oxidative stress. Circ Res 106:1712–1721
Goh YY, Pal M, Chong HC, Zhu P, Tan MJ, Punugu L, Lam CR, Yau YH, Tan CK, Huang RL, Tan SM, Tang MB, Ding JL, Kersten S, Tan NS (2010) Angiopoietin-like 4 interacts with integrins beta1 and beta5 to modulate keratinocyte migration. Am J Pathol 177:2791–2803
Grootaert C, Van de Wiele T, Van Roosbroeck I, Possemiers S, Vercoutter-Edouart AS, Verstraete W, Bracke M, Vanhoecke B (2011) Bacterial monocultures, propionate, butyrate and H2O2 modulate the expression, secretion and structure of the fasting-induced adipose factor in gut epithelial cell lines. Environ Microbiol 13:1778–1789
Hendy GN, Bevan S, Mattei MG, Mouland AJ (1995) Chromogranin A. Clin Invest Med 18:47–65
Jonker JT, Smit JW, Hammer S, Snel M, van der Meer RW, Lamb HJ, Mattijssen F, Mudde K, Jazet IM, Dekkers OM, de Roos A, Romijn JA, Kersten S, Rensen PC (2013) Dietary modulation of plasma angiopoietin-like protein 4 concentrations in healthy volunteers and in patients with type 2 diabetes. Am J Clin Nutr 97:255–260
Kersten S, Mandard S, Tan NS, Escher P, Metzger D, Chambon P, Gonzalez FJ, Desvergne B, Wahli W (2000) Characterization of the fasting-induced adipose factor FIAF, a novel peroxisome proliferator-activated receptor target gene. J Biol Chem 275:28488–28493
Kersten S, Lichtenstein L, Steenbergen E, Mudde K, Hendriks HF, Hesselink MK, Schrauwen P, Muller M (2009) Caloric restriction and exercise increase plasma ANGPTL4 levels in humans via elevated free fatty acids. Arterioscler Thromb Vasc Biol 29:969–974
Kim T, Tao-Cheng JH, Eiden LE, Loh YP (2001) Chromogranin A, an “on/off” switch controlling dense-core secretory granule biogenesis. Cell 106:499–509
Koliwad SK, Kuo T, Shipp LE, Gray NE, Backhed F, So AY, Farese RV Jr, Wang JC (2009) Angiopoietin-like 4 (ANGPTL4, fasting-induced adipose factor) is a direct glucocorticoid receptor target and participates in glucocorticoid-regulated triglyceride metabolism. J Biol Chem 284:25593–25601
Korecka A, de Wouters T, Cultrone A, Lapaque N, Pettersson S, Dore J, Blottiere HM, Arulampalam V (2013) ANGPTL4 expression induced by butyrate and rosiglitazone in human intestinal epithelial cells utilizes independent pathways. Am J Physiol Gastrointest Liver Physiol 304:G1025–G1037
Koster A, Chao YB, Mosior M, Ford A, Gonzalez-DeWhitt PA, Hale JE, Li D, Qiu Y, Fraser CC, Yang DD, Heuer JG, Jaskunas SR, Eacho P (2005) Transgenic angiopoietin-like (angptl)4 overexpression and targeted disruption of angptl4 and angptl3: regulation of triglyceride metabolism. Endocrinology 146:4943–4950
Lefebvre P, Cariou B, Lien F, Kuipers F, Staels B (2009) Role of bile acids and bile acid receptors in metabolic regulation. Physiol Rev 89:147–191
Lichtenstein L, Kersten S (2010) Modulation of plasma TG lipolysis by Angiopoietin-like proteins and GPIHBP1. Biochim Biophys Acta 1801:415–420
Lichtenstein L, Berbee JF, van Dijk SJ, van Dijk KW, Bensadoun A, Kema IP, Voshol PJ, Muller M, Rensen PC, Kersten S (2007) Angptl4 upregulates cholesterol synthesis in liver via inhibition of LPL- and HL-dependent hepatic cholesterol uptake. Arterioscler Thromb Vasc Biol 27:2420–2427
Lichtenstein L, Mattijssen F, de Wit NJ, Georgiadi A, Hooiveld GJ, van der Meer R, He Y, Qi L, Koster A, Tamsma JT, Tan NS, Muller M, Kersten S (2010) Angptl4 protects against severe proinflammatory effects of saturated fat by inhibiting fatty acid uptake into mesenteric lymph node macrophages. Cell Metab 12:580–592
Loh YP, Cheng Y, Mahata SK, Corti A, Tota B (2012) Chromogranin A and derived peptides in health and disease. J Mol Neurosc 48:347–356
Mandard S, Zandbergen F, Tan NS, Escher P, Patsouris D, Koenig W, Kleemann R, Bakker A, Veenman F, Wahli W, Muller M, Kersten S (2004) The direct peroxisome proliferator-activated receptor target fasting-induced adipose factor (FIAF/PGAR/ANGPTL4) is present in blood plasma as a truncated protein that is increased by fenofibrate treatment. J Biol Chem 279:34411–34420
Mandard S, Zandbergen F, van Straten E, Wahli W, Kuipers F, Muller M, Kersten S (2006) The fasting-induced adipose factor/Angiopoietin-like Protein 4 is physically associated with lipoproteins and governs plasma lipid levels and adiposity. J Biol Chem 281:934–944
Mellitzer G, Gradwohl G (2011) Enteroendocrine cells and lipid absorption. Curr Opin Lipidol 22:171–175
Moran GW, Leslie FC, Levison SE, Worthington J, McLaughlin JT (2008) Enteroendocrine cells: neglected players in gastrointestinal disorders? Therap Adv Gastroenterol 1:51–60
Murphy KG, Dhillo WS, Bloom SR (2006) Gut peptides in the regulation of food intake and energy homeostasis. Endocr Rev 27:719–727
Rindi G, Leiter AB, Kopin AS, Bordi C, Solcia E (2004) The “normal” endocrine cell of the gut: changing concepts and new evidences. Ann NY Acad Sci 1014:1–12
Steinert RE, Beglinger C (2011) Nutrient sensing in the gut: interactions between chemosensory cells, visceral afferents and the secretion of satiation peptides. Physiol Behav 105:62–70
Sukonina V, Lookene A, Olivecrona T, Olivecrona G (2006) Angiopoietin-like protein 4 converts lipoprotein lipase to inactive monomers and modulates lipase activity in adipose tissue. Proc Natl Acad Sci USA 103:17450–17455
Turnbaugh PJ, Backhed F, Fulton L, Gordon JI (2008) Diet-induced obesity is linked to marked but reversible alterations in the mouse distal gut microbiome. Cell Host Microbe 3:213–223
Xu A, Lam MC, Chan KW, Wang Y, Zhang J, Hoo RL, Xu JY, Chen B, Chow WS, Tso AW, Lam KS (2005) Angiopoietin-like protein 4 decreases blood glucose and improves glucose tolerance but induces hyperlipidemia and hepatic steatosis in mice. Proc Natl Acad Sci USA 102:6086–6091
Yau MH, Wang Y, Lam KS, Zhang J, Wu D, Xu A (2009) A highly conserved motif within the NH2-terminal coiled-coil domain of angiopoietin-like protein 4 confers its inhibitory effects on lipoprotein lipase by disrupting the enzyme dimerization. J Biol Chem 284:11942–11952
Yoon JC, Chickering TW, Rosen ED, Dussault B, Qin Y, Soukas A, Friedman JM, Holmes WE, Spiegelman BM (2000) Peroxisome proliferator-activated receptor gamma target gene encoding a novel angiopoietin-related protein associated with adipose differentiation. Mol Cell Biol 20:5343–5349
Yoshida K, Shimizugawa T, Ono M, Furukawa H (2002) Angiopoietin-like protein 4 is a potent hyperlipidemia-inducing factor in mice and inhibitor of lipoprotein lipase. J Lipid Res JID 0376606(43):1770–1772
Zhu P, Goh YY, Chin HF, Kersten S, Tan NS (2012) Angiopoietin-like 4: a decade of research. Biosci Rep 32:211–219
Acknowledgments
We would like to thank Dries Mulder for providing humans intestinal samples. Current address for Laeticia Lichtenstein: Institut National de la Santé et de la Recherche Médicale (INSERM), Unit 563, Toulouse, France. The work was partially funded by a Grant from the EFSD/MSD European Research Programme on Novel Therapies for Type 2 Diabetes.
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Alex, S., Lichtenstein, L., Dijk, W. et al. ANGPTL4 is produced by entero-endocrine cells in the human intestinal tract. Histochem Cell Biol 141, 383–391 (2014). https://doi.org/10.1007/s00418-013-1157-y
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DOI: https://doi.org/10.1007/s00418-013-1157-y