Display options
Cite
Reiter FP, Wimmer R, Wottke L, et al. Role of interleukin-1 and its antagonism of hepatic stellate cell proliferation and liver fibrosis in the Abcb4(-/-) mouse model. World J Hepatol. 2016;8(8):401-10doi: 10.4254/wjh.v8.i8.401.
Reiter, F. P., Wimmer, R., Wottke, L., Artmann, R., Nagel, J. M., Carranza, M. O., Mayr, D., Rust, C., Fickert, P., Trauner, M., Gerbes, A. L., Hohenester, S., & Denk, G. U. (2016). Role of interleukin-1 and its antagonism of hepatic stellate cell proliferation and liver fibrosis in the Abcb4(-/-) mouse model. World journal of hepatology, 8(8), 401-10. https://doi.org/10.4254/wjh.v8.i8.401
Reiter, Florian P, et al. "Role of interleukin-1 and its antagonism of hepatic stellate cell proliferation and liver fibrosis in the Abcb4(-/-) mouse model." World journal of hepatology vol. 8,8 (2016): 401-10. doi: https://doi.org/10.4254/wjh.v8.i8.401
Reiter FP, Wimmer R, Wottke L, Artmann R, Nagel JM, Carranza MO, Mayr D, Rust C, Fickert P, Trauner M, Gerbes AL, Hohenester S, Denk GU. Role of interleukin-1 and its antagonism of hepatic stellate cell proliferation and liver fibrosis in the Abcb4(-/-) mouse model. World J Hepatol. 2016 Mar 18;8(8):401-10. doi: 10.4254/wjh.v8.i8.401. PMID: 27004088; PMCID: PMC4794530.
Copy Download .nbib Format: NLM
Share it on

World J Hepatol. 2016 Mar 18;8(8):401-10. doi: 10.4254/wjh.v8.i8.401.

Role of interleukin-1 and its antagonism of hepatic stellate cell proliferation and liver fibrosis in the Abcb4(-/-) mouse model.

World journal of hepatology

Florian P Reiter, Ralf Wimmer, Lena Wottke, Renate Artmann, Jutta M Nagel, Manuel O Carranza, Doris Mayr, Christian Rust, Peter Fickert, Michael Trauner, Alexander L Gerbes, Simon Hohenester, Gerald U Denk

Affiliations

  1. Florian P Reiter, Ralf Wimmer, Lena Wottke, Renate Artmann, Jutta M Nagel, Alexander L Gerbes, Simon Hohenester, Gerald U Denk, Department of Medicine II, Liver Center Munich, University of Munich, D-81377 Munich, Germany.

PMID: 27004088 PMCID: PMC4794530 DOI: 10.4254/wjh.v8.i8.401

Abstract

AIM: To study the interleukin-1 (IL-1) pathway as a therapeutic target for liver fibrosis in vitro and in vivo using the ATP-binding cassette transporter b4(-/-) (Abcb4(-/-)) mouse model.

METHODS: Female and male Abcb4(-/-) mice from 6 to 13 mo of age were analysed for the degree of cholestasis (liver serum tests), extent of liver fibrosis (hydroxyproline content and Sirius red staining) and tissue-specific activation of signalling pathways such as the IL-1 pathway [quantitative polymerase chain reaction (qPCR)]. For in vivo experiments, murine hepatic stellate cells (HSCs) were isolated via pronase-collagenase perfusion followed by density gradient centrifugation using female mice. Murine HSCs were stimulated with up to 1 ng/mL IL-1β with or without 2.5 μg/mL Anakinra, an IL-1 receptor antagonist, respectively. The proliferation of murine HSCs was assessed via the BrdU assay. The toxicity of Anakinra was evaluated via the fluorescein diacetate hydrolysis (FDH) assay. In vivo 8-wk-old Abcb4(-/-) mice with an already fully established hepatic phenotype were treated with Anakinra (1 mg/kg body-weight daily intraperitoneally) or vehicle and liver injury and liver fibrosis were evaluated via serum tests, qPCR, hydroxyproline content and Sirius red staining.

RESULTS: Liver fibrosis was less pronounced in males than in female Abcb4(-/-) animals as defined by a lower hydroxyproline content (274 ± 64 μg/g vs 436 ± 80 μg/g liver, respectively; n = 13-15; P < 0.001; Mann-Whitney U-test) and lower mRNA expression of the profibrogenic tissue inhibitor of metalloproteinase-1 (TIMP) (1 ± 0.41 vs 0.66 ± 0.33 fold, respectively; n = 13-15; P < 0.05; Mann-Whitney U-test). Reduced liver fibrosis was associated with significantly lower levels of F4/80 mRNA expression (1 ± 0.28 vs 0.71 ± 0.41 fold, respectively; n = 12-15; P < 0.05; Mann-Whitney U-test) and significantly lower IL-1β mRNA expression levels (1 ± 0.38 vs 0.44 ± 0.26 fold, respectively; n = 13-15; P < 0.001; Mann-Whitney U-test). No gender differences in the serum liver parameters [bilirubin; alanine aminotransferase (ALT); aspartate aminotransferase and alkaline phosphatase (AP)] were found. In vitro, the administration of IL-1β resulted in a significant increase in HSC proliferation [0.94 ± 0.72 arbitrary units (A.U.) in untreated controls, 1.12 ± 0.80 A.U. at an IL-1β concentration of 0.1 ng/mL and 1.18 ± 0.73 A.U. at an IL-1β concentration of 1 ng/mL in samples from n = 6 donor animals; P < 0.001; analyses of variance (ANOVA)]. Proliferation was reduced significantly by the addition of 2.5 μg/mL Anakinra (0.81 ± 0.60 A.U. in untreated controls, 0.92 ± 0.68 A.U. at an IL-1β concentration of 0.1 ng/mL, and 0.91 ± 0.69 A.U. at an IL-1β concentration of 1 ng/mL; in samples from n = 6 donor animals; P < 0.001; ANOVA) suggesting an anti-proliferative effect of this clinically approved IL-1 receptor antagonist. The FDH assay showed this dose to be non-toxic in HSCs. In vivo, Anakinra had no effect on the hepatic hydroxyproline content, liver serum tests (ALT and AP) and pro-fibrotic (collagen 1α1, collagen 1α2, transforming growth factor-β, and TIMP-1) and anti-fibrotic [matrix metalloproteinase 2 (MMP2), MMP9 and MMP13] gene expression after 4 wk of treatment. Furthermore, the hepatic IL-1β and F4/80 mRNA expression levels were unaffected by Anakinra treatment.

CONCLUSION: IL-1β expression is associated with the degree of liver fibrosis in Abcb4(-/-) mice and promotes HSC proliferation. IL-1 antagonism shows antifibrotic effects in vitro but not in Abcb4(-/-) mice.

Keywords: Cholestasis; Interleukin-1; Liver fibrosis; Primary sclerosing cholangitis; The ATP-binding cassette transporter b4

References

  1. Lab Invest. 2004 Feb;84(2):153-9 - PubMed
  2. Am J Physiol Gastrointest Liver Physiol. 2009 Jun;296(6):G1324-31 - PubMed
  3. Hepatol Res. 2014 Dec;44(13):1286-98 - PubMed
  4. Nat Clin Pract Gastroenterol Hepatol. 2004 Dec;1(2):98-105 - PubMed
  5. Mod Pathol. 2007 Feb;20 Suppl 1:S15-30 - PubMed
  6. J Biol Chem. 2009 Jan 30;284(5):2908-16 - PubMed
  7. Hepatology. 2013 Dec;58(6):2045-55 - PubMed
  8. Gut. 2014 Nov;63(11):1782-92 - PubMed
  9. Hepat Med. 2011 Jul 04;3:69-80 - PubMed
  10. J Hepatol. 2012 Jul;57(1):133-40 - PubMed
  11. Intractable Rare Dis Res. 2015 Feb;4(1):1-6 - PubMed
  12. Gastroenterology. 2006 Feb;130(2):465-81 - PubMed
  13. Lab Invest. 2003 May;83(5):655-63 - PubMed
  14. APMIS. 2014 May;122(5):392-8 - PubMed
  15. Wien Med Wochenschr. 2008;158(19-20):542-8 - PubMed
  16. Biochem Biophys Res Commun. 2015 Apr 3;459(2):227-33 - PubMed
  17. J Clin Invest. 2012 Oct;122(10 ):3476-89 - PubMed
  18. Anal Biochem. 1987 Feb 15;161(1):207-18 - PubMed
  19. QJM. 2012 Sep;105(9):813-7 - PubMed
  20. Apoptosis. 2005 Oct;10(5):927-39 - PubMed
  21. Semin Liver Dis. 2010 Aug;30(3):245-57 - PubMed
  22. J Histochem Cytochem. 1985 Jan;33(1):77-9 - PubMed
  23. Clin Chim Acta. 1980 Jun 10;104(2):161-7 - PubMed
  24. Best Pract Res Clin Gastroenterol. 2011 Apr;25(2):195-206 - PubMed
  25. J Hepatol. 1997 Jan;26(1):138-45 - PubMed
  26. Gastroenterology. 1991 Jun;100(6):1710-7 - PubMed
  27. Am J Pathol. 1994 Nov;145(5):1237-45 - PubMed
  28. J Hepatol. 2010 Nov;53(5):918-26 - PubMed
  29. Hepatology. 2012 Jan;55(1):173-83 - PubMed
  30. J Hepatol. 2005 Dec;43(6):1045-54 - PubMed
  31. Hepatology. 2010 Feb;51(2):660-78 - PubMed
  32. J Biol Chem. 2000 Jan 28;275(4):2247-50 - PubMed
  33. J Hepatol. 2015 Apr;62(1 Suppl):S15-24 - PubMed
  34. Gastroenterology. 2004 Jul;127(1):261-74 - PubMed
  35. Physiol Rev. 2008 Jan;88(1):125-72 - PubMed
  36. Dig Liver Dis. 2002 Jun;34(6):387-92 - PubMed
  37. Circulation. 2008 May 20;117(20):2670-83 - PubMed

Publication Types