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Digestive Diseases and Sciences

, Volume 62, Issue 8, pp 2021–2034 | Cite as

Intestinal Alkaline Phosphatase Attenuates Alcohol-Induced Hepatosteatosis in Mice

  • Sulaiman R. Hamarneh
  • Byeong-Moo Kim
  • Kanakaraju Kaliannan
  • Sara A. Morrison
  • Tyler J. Tantillo
  • Qingsong Tao
  • Mussa M. Rafat Mohamed
  • Juan M. Ramirez
  • Aaron Karas
  • Wei Liu
  • Dong Hu
  • Abeba Teshager
  • Sarah Shireen Gul
  • Konstantinos P. Economopoulos
  • Atul K. Bhan
  • Madhu S. Malo
  • Michael Y. ChoiEmail author
  • Richard A. HodinEmail author
Original Article

Abstract

Background and Aims

Bacterially derived factors from the gut play a major role in the activation of inflammatory pathways in the liver and in the pathogenesis of alcoholic liver disease. The intestinal brush-border enzyme intestinal alkaline phosphatase (IAP) detoxifies a variety of bacterial pro-inflammatory factors and also functions to preserve gut barrier function. The aim of this study was to investigate whether oral IAP supplementation could protect against alcohol-induced liver disease.

Methods

Mice underwent acute binge or chronic ethanol exposure to induce alcoholic liver injury and steatosis ± IAP supplementation. Liver tissue was assessed for biochemical, inflammatory, and histopathological changes. An ex vivo co-culture system was used to examine the effects of alcohol and IAP treatment in regard to the activation of hepatic stellate cells and their role in the development of alcoholic liver disease.

Results

Pretreatment with IAP resulted in significantly lower serum alanine aminotransferase compared to the ethanol alone group in the acute binge model. IAP treatment attenuated the development of alcohol-induced fatty liver, lowered hepatic pro-inflammatory cytokine and serum LPS levels, and prevented alcohol-induced gut barrier dysfunction. Finally, IAP ameliorated the activation of hepatic stellate cells and prevented their lipogenic effect on hepatocytes.

Conclusions

IAP treatment protected mice from alcohol-induced hepatotoxicity and steatosis. Oral IAP supplementation could represent a novel therapy to prevent alcoholic-related liver disease in humans.

Keywords

Intestinal alkaline phosphatase Alcoholic liver disease Fatty liver Stellate cells 

Notes

Acknowledgments

Richard A. Hodin was supported by National Institute of Health grant NIH/NIDDK T32 (No. DK007754), The Ellison Foundation grant and Nutritional Obesity Research Center of Harvard (NORCH) NIH (No. P30-DK040561). We also thank the animal facility and the pathology lab at MGH for maintainance of animals and preparation of tissue sections.

Author's contribution

RAH, MYC, MSM, SRH, and BK contributed to study concept and theory; RAH, MYC, SRH, and BK contributed to research design; SRH, BK, KK, SAM, TJT, QT, MMRM, JMR, AK, WL, DH, AT, SSG, KPE, AKB, MSM, MYC, and RAH contributed to data acquisition; SRH, BK, KK, SAM, TJT, QT, MMRM, JMR, AK, WL, DH, AT, SSG, KPE, AKB, MSM, MYC, and RAH contributed to data analyses and interpretation; SRH, BK, SAM, KPE, MYC, and RAH contributed to statistical analyses; SRH, BK, CYM, and RAH contributed to drafting of the manuscript; all authors contributed to critical review of the manuscript for important intellectual content; RAH obtained funding; all authors contributed to approval of the manuscript; RAH and MYC supervised the study.

Compliance with ethical standards

Conflict of interest

None.

References

  1. 1.
    Miniño AM, Murphy SL, Xu J, et al. Deaths: final data for 2008. Natl Vital Stat Rep. 2011;7:1–126.Google Scholar
  2. 2.
    Varma V, Webb K, Mirza DF. Liver transplantation for alcoholic liver disease. World J Gastroenterol. 2010;21:4377–4393.CrossRefGoogle Scholar
  3. 3.
    O’Shea RS, Dasarathy S, McCullough AJ. Alcoholic liver disease. Hepatology. 2010;51:307–328.CrossRefPubMedGoogle Scholar
  4. 4.
    Celli R, Zhang X. Pathology of alcoholic liver disease. J Clin Transl Hepatol. 2014;15:103–109.Google Scholar
  5. 5.
    Thurman RG. Alcoholic liver injury involves activation of Kupffer cells by endotoxin. Am J Physiol Gastrointest Liver Physiol. 1998;275:G605–G611.Google Scholar
  6. 6.
    Lin HZ, Yang SQ, Zeldin G, et al. Chronic ethanol consumption induces the production of tumor necrosis factor-alpha and related cytokines in liver and adipose tissue. Alcohol Clin Exp Res. 1998;22:231S–237S.CrossRefPubMedGoogle Scholar
  7. 7.
    Inokuchi S, Tsukamoto H, Park E, et al. Toll-like receptor 4 mediates alcohol-induced steatohepatitis through bone marrow-derived and endogenous liver cells in mice. Alcohol Clin Exp Res. 2011;35:1509–1518.PubMedPubMedCentralGoogle Scholar
  8. 8.
    Bird GL, Sheron N, Goka AK, et al. Increased plasma tumor necrosis factor in sever alcoholic hepatitis. Ann Intern Med. 1990;112:917–920.CrossRefPubMedGoogle Scholar
  9. 9.
    Felver ME, Mezey E, McGuire M, et al. Plasma tumor necrosis factor alpha predicts decreased long term survival in severe alcoholic hepatitis. Alcohol Clin Exp Res. 1990;14:255–259.CrossRefPubMedGoogle Scholar
  10. 10.
    Seki E, Brenner DA. Toll-like receptors and adaptor molecules in liver disease: update. Hepatology. 2008;48:322–335.CrossRefPubMedGoogle Scholar
  11. 11.
    Petrasek J, Mandrekar P, Szabo G. Toll-like receptors in the pathogenesis of alcoholic liver disease. Gastroenterol Res Pract. 2010;2010:710381.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Nanji AA, Khettry U, Sadrzadeh SM, et al. Severity of liver injury in experimental alcoholic liver disease. Correlation with plasma endotoxin, prostaglandin E2, leukotriene B4, and thromboxane B2. Am J Pathol. 1993;142:367–373.PubMedPubMedCentralGoogle Scholar
  13. 13.
    Fukui H. Relation of endotoxin, endotoxin binding proteins and macrophages to severe alcoholic liver injury and multiple organ failure. Alcohol Clin Exp Res. 2005;29:172S–179S.CrossRefPubMedGoogle Scholar
  14. 14.
    Horie Y, Kato S, Ohki E, et al. Role of endothelin in endotoxin-induced hepatic microvascular dysfunction in rats fed chronically with ethanol. J Gastroenterol Hepatol. 2001;16:916–922.CrossRefPubMedGoogle Scholar
  15. 15.
    Horie Y, Kato S, Ohki E, et al. Hepatic microvascular dysfunction in endotoxemic rats after acute ethanol administration. Alcohol Clin Exp Res. 2000;24:691–698.CrossRefPubMedGoogle Scholar
  16. 16.
    Deaciuc IV, Nikolova-Karakashian M, Fortunato F, et al. Apoptosis and dysregulated ceramide metabolism in a murine model of alcohol enhanced lipopolysaccharide hepatotoxicity. Alcohol Clin Exp Res. 2000;24:1557–1565.CrossRefPubMedGoogle Scholar
  17. 17.
    Sandahl TD, Grønbaek H, Møller HJ, et al. Hepatic macrophage activation and the LPS pathway in patients with alcoholic hepatitis: a prospective cohort study. Am J Gastroenterol. 2014;109:1749–1756.CrossRefPubMedGoogle Scholar
  18. 18.
    Mathurin P, Deng QG, Keshavarzian A, et al. Exacerbation of alcoholic liver injury by enteral endotoxin in rats. Hepatology. 2000;32:1008–1017.CrossRefPubMedGoogle Scholar
  19. 19.
    Malo MS, Alam SN, Mostafa G, et al. Intestinal alkaline phosphatase preserves the normal homeostasis of gut microbiota. Gut. 2010;59:1476–1484.CrossRefPubMedGoogle Scholar
  20. 20.
    Chen KT, Malo MS, Beasley-Topliffe LK, et al. A role for intestinal alkaline phosphatase in the maintenance of local gut immunity. Dig Dis Sci. 2011;56:1020–1027.CrossRefPubMedGoogle Scholar
  21. 21.
    Bates JM, Akerlund J, Mittge E, et al. Intestinal alkaline phosphatase detoxifies lipopolysaccharide and prevents inflammation in zebrafish in response to the gut microbiota. Cell Host Microbe. 2007;2:371–382.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Chen KT, Malo MS, Moss AK, et al. Identification of specific targets for the gut mucosal defense factor intestinal alkaline phosphatase. Am J Physiol Gastrointest Liver Physiol. 2010;299:G467–G475.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Rentea RM, Liedel JL, Welak SR, et al. Intestinal alkaline phosphatase administration in newborns is protective of gut barrier function in a neonatal necrotizing enterocolitis rat model. J Pediatr Surg. 2012;47:1135–1142.CrossRefPubMedGoogle Scholar
  24. 24.
    Nakano T, Inoue I, Koyama I, et al. Disruption of the murine intestinal alkaline phosphatase gene Akp3 impairs lipid transcytosis and induces visceral fat accumulation and hepatic steatosis. Am J Physiol Gastrointest Liver Physiol. 2007;292:G1439–G1449.CrossRefPubMedGoogle Scholar
  25. 25.
    Kaliannan K, Hamarneh SR, Economopoulos KP, et al. Intestinal alkaline phosphatase prevents metabolic syndrome in mice. Proc Natl Acad Sci USA. 2013;110:7003–7008.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Hufnagel H, Bode C, Bode JC, et al. Damage of rat small intestine induced by ethanol. Effect of ethanol on fecal excretion of intestinal alkaline phosphatase. Res Exp Med. 1980;178:65–70.CrossRefGoogle Scholar
  27. 27.
    Eloy R, Battinger F, Bignon JY, et al. Intestinal brush border enzymes and chronic alcohol ingestion. Res Exp Med. 1979;175:257–269.CrossRefGoogle Scholar
  28. 28.
    Iakovleva LM. Structural and functional characteristic of rat jejunum after long-term exposure to alcohol. Morfologiia. 2012;141:45–48.PubMedGoogle Scholar
  29. 29.
    Carson EJ, Pruett SB. Development and characterization of a binge drinking model in mice for evaluation of the immunological effects of ethanol. Alcohol Clin Exp Res. 1996;20:132–138.CrossRefPubMedGoogle Scholar
  30. 30.
    Zhou Z, Sun X, Lambert JC, et al. Metallothionein-independent zinc protection from alcoholic liver injury. Am J Pathol. 2002;160:2267–2274.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Folch J, Lees M. SLOANE STANLEY GH. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957;226:497–509.PubMedGoogle Scholar
  32. 32.
    Mohar I, Brempelis KJ, Murray SA, et al. Isolation of non-parenchymal cells from the mouse liver. Methods Mol Biol. 2015;1325:3–17.CrossRefPubMedGoogle Scholar
  33. 33.
    Hamarneh S, Morrison SA, Tantillo TJ, et al. A novel approach to maintaining gut mucosal integrity using an oral enzyme supplement. Ann Surg. 2014;260:706–715.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Reeves HL, Burt AD, Wood S, et al. Hepatic stellate cell activation occurs in the absence of hepatitis in alcoholic liver disease and correlates with the severity of steatosis. J Hepatol. 1996;25:677–683.CrossRefPubMedGoogle Scholar
  35. 35.
    Jeong WI, Osei-Hyiaman D, Park O, et al. Paracrine activation of hepatic CB1 receptors by stellate cell-derived endocannabinoids mediates alcoholic fatty liver. Cell Metab. 2008;7:227–235.CrossRefPubMedGoogle Scholar
  36. 36.
    Fukui H, Brauner B, Bode JC, et al. Plasma endotoxin concentrations in patients with alcoholic and nonalcoholic liver disease: reevaluation with an improved chromogenic assay. J Hepatol. 1991;12:162–169.CrossRefPubMedGoogle Scholar
  37. 37.
    Bode C, Fukui H, Bode JC. “Hidden” endotoxin in plasma of patients with alcoholic liver disease. Eur J Gastroenterol Hepatol. 1993;5:257–262.CrossRefGoogle Scholar
  38. 38.
    Rao R. Endotoxemia and gut barrier dysfunction in alcoholic liver disease. Hepatology. 2009;50:638–644.CrossRefPubMedGoogle Scholar
  39. 39.
    Tamai H, Kato S, Horie Y, et al. Effect of acute ethanol administration on the intestinal absorption of endotoxin in rats. Alcohol Clin Exp Res. 2000;24:390–394.CrossRefPubMedGoogle Scholar
  40. 40.
    Yan AW, Fouts DE, Brandl J, et al. Enteric dysbiosis associated with a mouse model of alcoholic liver disease. Hepatology. 2011;53:96–105.CrossRefPubMedGoogle Scholar
  41. 41.
    Adachi Y, Moore LE, Bradford BU, et al. Antibiotics prevent liver injury in rats following long-term exposure to ethanol. Gastroenterology. 1995;108:218–224.CrossRefPubMedGoogle Scholar
  42. 42.
    Forsyth CB, Farhadi A, Jakate SM, et al. Lactobacillus GG treatment ameliorates alcohol-induced intestinal oxidative stress, gut leakiness, and liver injury in a rat model of alcoholic steatohepatitis. Alcohol. 2009;43:163–172.CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Eliakim R, Mahmood A, Alpers DH. Rat intestinal alkaline phosphatase secretion into lumen and serum is coordinately regulated. Biochim Biophys Acta. 1991;1091:1–8.CrossRefPubMedGoogle Scholar
  44. 44.
    Schaller K, Höller H. Thiamine absorption in the rat. II. Intestinal alkaline phosphatase activity and thiamine absorption from rat small intestine in vitro and in-vivo. Int J Vitam Nutr Res. 1975;45:30–38.PubMedGoogle Scholar
  45. 45.
    Tsukamoto H, Lu SC. Current concepts in the pathogenesis of alcoholic liver injury. FASEB J. 2001;15:1335–1349.CrossRefPubMedGoogle Scholar
  46. 46.
    Goldberg RF, Austen WG Jr, Zhang X, et al. Intestinal alkaline phosphatase is a gut mucosal defense factor maintained by enteral nutrition. Proc Natl Acad Sci USA. 2008;4:3551–3556.CrossRefGoogle Scholar
  47. 47.
    Watson AJ, Hughes KR. TNF-α-induced intestinal epithelial cell shedding: implications for intestinal barrier function. Ann N Y Acad Sci. 2012;1258:1–8.CrossRefPubMedGoogle Scholar
  48. 48.
    Gustot T, Lemmers A, Moreno C, et al. Differential liver sensitization to toll-like receptor pathways in mice with alcoholic fatty liver. Hepatology. 2006;43:989–1000.CrossRefPubMedGoogle Scholar
  49. 49.
    Wang Z, Wu X, Zhang Y, et al. Discrepant roles of CpG ODN on acute alcohol-induced liver injury in mice. Int Immunopharmacol. 2012;12:526–533.CrossRefPubMedGoogle Scholar
  50. 50.
    Oak S, Mandrekar P, Catalano D, et al. TLR2- and TLR4-mediated signals determine attenuation or augmentation of inflammation by acute alcohol in monocytes. J Immunol. 2006;176:7628–7635.CrossRefPubMedGoogle Scholar
  51. 51.
    Lallès JP. Intestinal alkaline phosphatase: novel functions and protective effects. Nutr Rev. 2013;72:82–94.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Sulaiman R. Hamarneh
    • 1
  • Byeong-Moo Kim
    • 2
  • Kanakaraju Kaliannan
    • 1
  • Sara A. Morrison
    • 1
  • Tyler J. Tantillo
    • 1
  • Qingsong Tao
    • 1
  • Mussa M. Rafat Mohamed
    • 1
  • Juan M. Ramirez
    • 1
  • Aaron Karas
    • 1
  • Wei Liu
    • 1
  • Dong Hu
    • 1
  • Abeba Teshager
    • 1
  • Sarah Shireen Gul
    • 1
  • Konstantinos P. Economopoulos
    • 1
  • Atul K. Bhan
    • 3
  • Madhu S. Malo
    • 1
  • Michael Y. Choi
    • 2
    • 4
    Email author
  • Richard A. Hodin
    • 1
    Email author
  1. 1.Department of Surgery, Harvard Medical SchoolMassachusetts General HospitalBostonUSA
  2. 2.Gastrointestinal Unit, Department of Medicine, Harvard Medical SchoolMassachusetts General HospitalBostonUSA
  3. 3.Department of Pathology, Harvard Medical SchoolMassachusetts General HospitalBostonUSA
  4. 4.Harvard Stem Cell InstituteCambridgeUSA

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