Abstract
Phytosterols, components of soy-derived lipids, are among the proposed exacerbants of parenteral nutrition–associated cholestasis (PNAC). We investigated whether phytosterols contribute to bile acid (BA)–induced hepatocyte damage by antagonizing a nuclear receptor (NR) critically involved in hepatoprotection from cholestasis, FXR (farnesoid X receptor, NR1H4). In HepG2 cells, stigmasterol acetate (StigAc), a water-soluble Stig derivative, suppressed ligand-activated expression of FXR target genes involved in adaptation to cholestasis (i.e. BSEP, FGF-19, OSTα/β). Furthermore, StigAc antagonized BA-activated, FXR target genes SHP and BSEP in FXR+/+, but not in FXR−/− mouse hepatocytes. Both Stig and StigAc inhibited BA-activated, FXR-dependent reporter gene expression in transfected HepG2 cells, whereas the most prevalent phytosterol in lipids, β-sitosterol, had no inhibitory effect. Finally, among six ligand-activated NR-ligand binding domains (LBDs) tested, antagonism by StigAc was specific to only two (FXR and PXR, pregnane X receptor, NR1I2). We demonstrate that Stig, a phytosterol prevalent in soy-derived PN lipid solutions, is a potent in vitro antagonist of the NR for bile acids FXR.
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Abbreviations
- BA:
-
bile acid
- BSEP:
-
bile salt export pump
- CDCA:
-
chenodeoxycholic acid
- FGF:
-
fibroblast growth factor
- FXR:
-
farnesoid X receptor
- LBD:
-
ligand-binding domain
- LXR:
-
liver X receptor
- NR:
-
nuclear receptor
- NTCP:
-
sodium taurocholate cotransporting polypeptide
- OST:
-
organic solute transporter
- PN (AC):
-
parenteral nutrition (–associated cholestasis)
- PPARγ:
-
peroxisome proliferator-activated receptor gamma
- PXR:
-
pregnane X receptor
- RXR:
-
retinoid X receptor
- SHP:
-
small heterodimer partner
- Stig (Ac):
-
stigmasterol (acetate)
- VDR:
-
vitamin D receptor
References
Kelly DA 1998 Liver complications of pediatric parenteral nutrition–epidemiology. Nutrition 14: 153–157
Teitelbaum DH, Tracy T 2001 Parenteral nutrition-associated cholestasis. Semin Pediatr Surg 10: 72–80
Btaiche IF, Khalidi N 2002 Parenteral nutrition-associated liver complications in children. Pharmacotherapy 22: 188–211
Ostlund RE Jr 2002 Phytosterols in human nutrition. Annu Rev Nutr 22: 533–549
Ostlund RE Jr, McGill JB, Zeng CM, Covey DF, Stearns J, Stenson WF, Spilburg CA 2002 Gastrointestinal absorption and plasma kinetics of soy Delta(5)-phytosterols and phytostanols in humans. Am J Physiol Endocrinol Metab 282: E911–E916
Repa JJ, Berge KE, Pomajzl C, Richardson JA, Hobbs H, Mangelsdorf DJ 2002 Regulation of ATP-binding cassette sterol transporters ABCG5 and ABCG8 by the liver X receptors alpha and beta. J Biol Chem 277: 18793–18800
Van Aerde JE, Duerksen DR, Gramlich L, Meddings JB, Chan G, Thomson AB, Clandinin MT 1999 Intravenous fish oil emulsion attenuates total parenteral nutrition-induced cholestasis in newborn piglets. Pediatr Res 45: 202–208
Iyer KR, Spitz L, Clayton P 1998 BAPS prize lecture: New insight into mechanisms of parenteral nutrition-associated cholestasis: role of plant sterols. British Association of Paediatric Surgeons. J Pediatr Surg 33: 1–6
Clayton PT, Bowron A, Mills KA, Massoud A, Casteels M, Milla PJ 1993 Phytosterolemia in children with parenteral nutrition-associated cholestatic liver disease. Gastroenterology 105: 1806–1813
Wang H, Chen J, Hollister K, Sowers LC, Forman BM 1999 Endogenous bile acids are ligands for the nuclear receptor FXR/BAR. Mol Cell 3: 543–553
Trauner M, Boyer JL 2003 Bile salt transporters: molecular characterization, function, and regulation. Physiol Rev 83: 633–671
Boyer JL, Trauner M, Mennone A, Soroka CJ, Cai SY, Moustafa T, Zollner G, Lee JY, Ballatori N 2006 Upregulation of a basolateral FXR-dependent bile acid efflux transporter OSTalpha-OSTbeta in cholestasis in humans and rodents. Am J Physiol Gastrointest Liver Physiol 290: G1124–G1130
Goodwin B, Jones SA, Price RR, Watson MA, McKee DD, Moore LB, Galardi C, Wilson JG, Lewis MC, Roth ME, Maloney PR, Willson TM, Kliewer SA 2000 A regulatory cascade of the nuclear receptors FXR, SHP-1, and LRH-1 represses bile acid biosynthesis. Mol Cell 6: 517–526
Karpen SJ 2002 Nuclear receptor regulation of hepatic function. J Hepatol 36: 832–850
Zollner G, Wagner M, Moustafa T, Fickert P, Silbert D, Gumhold J, Fuchsbichler A, Halilbasic E, Denk H, Marschall HU, Trauner M 2006 Coordinated induction of bile acid detoxification and alternative elimination in mice: role of FXR-regulated organic solute transporter-alpha/beta in the adaptive response to bile acids. Am J Physiol Gastrointest Liver Physiol 290: G923–G932
Sinal CJ, Tohkin M, Miyata M, Ward JM, Lambert G, Gonzalez FJ 2000 Targeted disruption of the nuclear receptor FXR/BAR impairs bile acid and lipid homeostasis. Cell 102: 731–744
Liu Y, Binz J, Numerick MJ, Dennis S, Luo G, Desai B, MacKenzie KI, Mansfield TA, Kliewer SA, Goodwin B, Jones SA 2003 Hepatoprotection by the farnesoid X receptor agonist GW4064 in rat models of intra- and extrahepatic cholestasis. J Clin Invest 112: 1678–1687
Sokol RJ 1997 Total parenteral nutrition-related liver disease. Zhonghua Min Guo Xiao Er Ke Yi Xue Hui Za Zhi 38: 418–428
Berry MN, Friend DS 1969 High-yield preparation of isolated rat liver parenchymal cells: a biochemical and fine structural study. J Cell Biol 43: 506–520
Plat J, Brzezinka H, Lutjohann D, Mensink RP, von Bergmann K 2001 Oxidized plant sterols in human serum and lipid infusions as measured by combined gas-liquid chromatography-mass spectrometry. J Lipid Res 42: 2030–2038
Landrier JF, Eloranta JJ, Vavricka SR, Kullak-Ublick GA 2006 The nuclear receptor for bile acids, FXR, transactivates human organic solute transporter-alpha and -beta genes. Am J Physiol Gastrointest Liver Physiol 290: G476–G485
Holt JA, Luo G, Billin AN, Bisi J, McNeill YY, Kozarsky KF, Donahee M, Wang DY, Mansfield TA, Kliewer SA, Goodwin B, Jones SA 2003 Definition of a novel growth factor-dependent signal cascade for the suppression of bile acid biosynthesis. Genes Dev 17: 1581–1591
Ananthanarayanan M, Balasubramanian N, Makishima M, Mangelsdorf DJ, Suchy FJ 2001 Human bile salt export pump promoter is transactivated by the farnesoid X receptor/bile acid receptor. J Biol Chem 276: 28857–28865
Colomb V, Jobert-Giraud A, Lacaille F, Goulet O, Fournet JC, Ricour C 2000 Role of lipid emulsions in cholestasis associated with long-term parenteral nutrition in children. JPEN J Parenter Enteral Nutr 24: 345–350
Salen G, Horak I, Rothkopf M, Cohen JL, Speck J, Tint GS, Shore V, Dayal B, Chen T, Shefer S 1985 Lethal atherosclerosis associated with abnormal plasma and tissue sterol composition in sitosterolemia with xanthomatosis. J Lipid Res 26: 1126–1133
Bhattacharyya AK, Connor WE 1974 Beta-sitosterolemia and xanthomatosis. A newly described lipid storage disease in two sisters. J Clin Invest 53: 1033–1043
Klett EL, Lee MH, Adams DB, Chavin KD, Patel SB 2004 Localization of ABCG5 and ABCG8 proteins in human liver, gall bladder and intestine. BMC Gastroenterol 4: 21
Berge KE, Tian H, Graf GA, Yu L, Grishin NV, Schultz J, Kwiterovich P, Shan B, Barnes R, Hobbs HH 2000 Accumulation of dietary cholesterol in sitosterolemia caused by mutations in adjacent ABC transporters. Science 290: 1771–1775
Miettinen TA, Klett EL, Gylling H, Isoniemi H, Patel SB 2006 Liver transplantation in a patient with sitosterolemia and cirrhosis. Gastroenterology 130: 542–547
Tomer G, Ananthanarayanan M, Weymann A, Balasubramanian N, Suchy FJ 2003 Differential developmental regulation of rat liver canalicular membrane transporters Bsep and Mrp2. Pediatr Res 53: 288–294
Kearns GL, Abdel-Rahman SM, Alander SW, Blowey DL, Leeder JS, Kauffman RE 2003 Developmental pharmacology–drug disposition, action, and therapy in infants and children. N Engl J Med 349: 1157–1167
Chen HL, Chen HL, Liu YJ, Feng CH, Wu CY, Shyu MK, Yuan RH, Chang MH 2005 Developmental expression of canalicular transporter genes in human liver. J Hepatol 43: 472–477
Moseley RH, Wang W, Takeda H, Lown K, Shick L, Ananthanarayanan M, Suchy FJ 1996 Effect of endotoxin on bile acid transport in rat liver: a potential model for sepsis-associated cholestasis. Am J Physiol 271: G137–G146
Ghose R, Zimmerman TL, Thevananther S, Karpen SJ 2004 Endotoxin leads to rapid subcellular re-localization of hepatic RXRalpha: a novel mechanism for reduced hepatic gene expression in inflammation. Nucl Recept 2: 4
Zimmerman TL, Thevananther S, Ghose R, Burns AR, Karpen SJ 2006 Nuclear export of retinoid X receptor {alpha} in response to interleukin-1beta-mediated cell signaling: roles for JNK and SER260. J Biol Chem 281: 15434–15440
van Mil SW, van der Woerd WL, van der Brugge G, Sturm E, Jansen PL, Bull LN, van den Berg IE, Berger R, Houwen RH, Klomp LW 2004 Benign recurrent intrahepatic cholestasis type 2 is caused by mutations in ABCB11. Gastroenterology 127: 379–384
Nishimura T, Utsunomiya Y, Hoshikawa M, Ohuchi H, Itoh N 1999 Structure and expression of a novel human FGF, FGF-19, expressed in the fetal brain. Biochim Biophys Acta 1444: 148–151
Inagaki T, Choi M, Moschetta A, Peng L, Cummins CL, McDonald JG, Luo G, Jones SA, Goodwin B, Richardson JA, Gerard RD, Repa JJ, Mangelsdorf DJ, Kliewer SA 2005 Fibroblast growth factor 15 functions as an enterohepatic signal to regulate bile acid homeostasis. Cell Metab 2: 217–225
Xie W, Radominska-Pandya A, Shi Y, Simon CM, Nelson MC, Ong ES, Waxman DJ, Evans RM 2001 An essential role for nuclear receptors SXR/PXR in detoxification of cholestatic bile acids. Proc Natl Acad Sci U S A 98: 3375–3380
Shih DM, Kast-Woelbern HR, Wong J, Xia YR, Edwards PA, Lusis AJ 2006 A role for FXR and human FGF-19 in the repression of paraoxonase-1 gene expression by bile acids. J Lipid Res 47: 384–392
Acknowledgements
The authors gratefully appreciate the expertise at the Analyte Center at Baylor College of Medicine (Dr. William E. O'Brien, Ph.D., Director) for accurate measurements of phytosterol concentrations, and members of the Karpen Laboratory for their input into the design and execution of experiments.
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Supported in part by National Institutes of Health Grant K12HD41648 and the American Gastroenterological Association/Roche Research Scholar Award in Liver Disease (B.A.C.), DK56239 (S.J.K.), and the Texas Gulf Coast Digestive Disease Center (DK56338).
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Carter, B., Taylor, O., Prendergast, D. et al. Stigmasterol, a Soy Lipid–Derived Phytosterol, Is an Antagonist of the Bile Acid Nuclear Receptor FXR. Pediatr Res 62, 301–306 (2007). https://doi.org/10.1203/PDR.0b013e3181256492
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DOI: https://doi.org/10.1203/PDR.0b013e3181256492
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