Skip to main content

Advertisement

SpringerLink
Log in

Von Willebrand factor protects against acute CCl4-induced hepatotoxicity through phospho-p38 MAPK signaling pathway inhibition

  • Original Article
  • Published:
Immunologic Research Aims and scope Submit manuscript

Abstract

The blood glycoprotein von Willebrand factor (vWF) is involved in coagulopathy and inflammation; however, its role in the pathogenesis of acute liver failure, as suggested by its higher expression levels in such patients, remains unknown. In this study, vWF-knockout (KO) mice showed more severe carbon tetrachloride (CCl4)-induced liver injury than wild-type mice. Patients with acute liver injury also showed elevated vWF protein activity and expression in liver tissues, as compared to healthy individuals. Using the mouse model and cultured human umbilical vein endothelial cells (HUVECs), CCl4 was found to directly increase vWF protein expression through interaction with the highly expressed vWF receptor, GPIbα. Microarray analysis revealed that the genes showing the most differential expression in response to CCl4-induced liver injury and vWF deficiency were related to the MAPK signaling pathway. Subsequent inhibition of vWF protein activity in HUVECs led to activation of the MAPK signal pathway and elevated production of FGL2, and treatment with a phospho-p38 inhibitor suppressed the CCl4-induced production of FGL2. Exposure of liver sinusoidal endothelial cells isolated from the vWF-KO acute liver injury model mice to phospho-p38 inhibitor also decreased FGL2 expression. The vWF/GPIbα axis plays a protective role against development of acute liver injury by attenuating FGL2 production through the MAPK signaling pathway. Collectively, these data provide insight into the pathogenesis of acute liver injury and a potential novel strategy for its treatment.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. McAvoy N, Thomson E, Wilson ES. Hepatic failure. Anaesth Intensive Care Med. 2012;13(4):161–5.

    Article  Google Scholar 

  2. Polson J, Lee WM. American Association for the Study of Liver D. AASLD position paper: the management of acute liver failure. Hepatology. 2005;41(5):1179–97.

    Article  PubMed  Google Scholar 

  3. Habib M, Roberts LN, Patel RK, Wendon J, Bernal W, Arya R. Evidence of rebalanced coagulation in acute liver injury and acute liver failure as measured by thrombin generation. Liver Int. 2014;34(5):672–8.

    Article  CAS  PubMed  Google Scholar 

  4. De Ceunynck K, De Meyer SF, Vanhoorelbeke K. Unwinding the von Willebrand factor strings puzzle. Blood. 2013;121(2):270–7.

    Article  PubMed  Google Scholar 

  5. Lenting PJ, VANS CJ, Denis CV. Clearance mechanisms of von Willebrand factor and factor VIII. J Thromb Haemost. 2007;5(7):1353–60.

    Article  CAS  PubMed  Google Scholar 

  6. Luo GP, Ni B, Yang X, Wu YZ. von Willebrand factor: more than a regulator of hemostasis and thrombosis. Acta Haematol. 2012;128(3):158–69.

    Article  CAS  PubMed  Google Scholar 

  7. Lenting PJ, Westein E, Terraube V, Ribba AS, Huizinga EG, Meyer D, et al. An experimental model to study the in vivo survival of von Willebrand factor. Basic aspects and application to the R1205H mutation. J Biol Chem. 2004;279(13):12102–9.

    Article  CAS  PubMed  Google Scholar 

  8. van Schooten CJ, Shahbazi S, Groot E, Oortwijn BD, van den Berg HM, Denis CV, et al. Macrophages contribute to the cellular uptake of von Willebrand factor and factor VIII in vivo. Blood. 2008;112(5):1704–12.

    Article  PubMed  Google Scholar 

  9. Petri B, Broermann A, Li H, Khandoga AG, Zarbock A, Krombach F, et al. von Willebrand factor promotes leukocyte extravasation. Blood. 2010;116(22):4712–9.

    Article  CAS  PubMed  Google Scholar 

  10. Zheng N, Shi X, Chen X, Lv W. Associations between inflammatory markers, hemostatic markers, and microvascular complications in 182 Chinese patients with type 2 diabetes mellitus. Lab Med. 2015;46(3):214–20.

    Article  PubMed  Google Scholar 

  11. Reuken PA, Kussmann A, Kiehntopf M, Budde U, Stallmach A, Claus RA, et al. Imbalance of von Willebrand factor and its cleaving protease ADAMTS13 during systemic inflammation superimposed on advanced cirrhosis. Liver Int. 2015;35(1):37–45.

    Article  CAS  PubMed  Google Scholar 

  12. Hugenholtz GC, Adelmeijer J, Meijers JC, Porte RJ, Stravitz RT, Lisman T. An unbalance between von Willebrand factor and ADAMTS13 in acute liver failure: implications for hemostasis and clinical outcome. Hepatology. 2013;58(2):752–61.

    Article  CAS  PubMed  Google Scholar 

  13. Kelley D, Lester C, Shaw S, de Laforcade A, Webster CR. Thromboelastographic evaluation of dogs with acute liver disease. J Vet Intern Med. 2015;29(4):1053–62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Yan J, Kong LY, Hu J, Gabrusiewicz K, Dibra D, Xia X et al. FGL2 as a multimodality regulator of tumor-mediated immune suppression and therapeutic target in gliomas. J Natl Cancer Inst. 2015;107(8).

  15. Marazzi S, Blum S, Hartmann R, Gundersen D, Schreyer M, Argraves S, et al. Characterization of human fibroleukin, a fibrinogen-like protein secreted by T lymphocytes. J Immunol. 1998;161(1):138–47.

    CAS  PubMed  Google Scholar 

  16. Ghanekar A, Mendicino M, Liu H, He W, Liu M, Zhong R, et al. Endothelial induction of fgl2 contributes to thrombosis during acute vascular xenograft rejection. J Immunol. 2004;172(9):5693–701.

    Article  CAS  PubMed  Google Scholar 

  17. Liu J, Tan Y, Zhang J, Zou L, Deng G, Xu X, et al. C5aR, TNF-alpha, and FGL2 contribute to coagulation and complement activation in virus-induced fulminant hepatitis. J Hepatol. 2015;62(2):354–62.

    Article  CAS  PubMed  Google Scholar 

  18. Xu GL, Chen J, Yang F, Li GQ, Zheng LX, Wu YZ. C5a/C5aR pathway is essential for the pathogenesis of murine viral fulminant hepatitis by way of potentiating Fgl2/fibroleukin expression. Hepatology. 2014;60(1):114–24.

    Article  CAS  PubMed  Google Scholar 

  19. Liu M, Mendicino M, Ning Q, Ghanekar A, He W, McGilvray I, et al. Cytokine-induced hepatic apoptosis is dependent on FGL2/fibroleukin: the role of Sp1/Sp3 and STAT1/PU.1 composite cis elements. J Immunol. 2006;176(11):7028–38.

    Article  CAS  PubMed  Google Scholar 

  20. Hancock WW, Szaba FM, Berggren KN, Parent MA, Mullarky IK, Pearl J, et al. Intact type 1 immunity and immune-associated coagulative responses in mice lacking IFN gamma-inducible fibrinogen-like protein 2. Proc Natl Acad Sci U S A. 2004;101(9):3005–10.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Selzner N, Liu H, Boehnert MU, Adeyi OA, Shalev I, Bartczak AM, et al. FGL2/fibroleukin mediates hepatic reperfusion injury by induction of sinusoidal endothelial cell and hepatocyte apoptosis in mice. J Hepatol. 2012;56(1):153–9.

    Article  CAS  PubMed  Google Scholar 

  22. Liu M, Leibowitz JL, Clark DA, Mendicino M, Ning Q, Ding JW, et al. Gene transcription of fgl2 in endothelial cells is controlled by Ets-1 and Oct-1 and requires the presence of both Sp1 and Sp3. Eur J Biochem. 2003;270(10):2274–86.

    Article  CAS  PubMed  Google Scholar 

  23. Pritchard MT, Cohen JI, Roychowdhury S, Pratt BT, Nagy LE. Early growth response-1 attenuates liver injury and promotes hepatoprotection after carbon tetrachloride exposure in mice. J Hepatol. 2010;53(4):655–62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Yang X, Sun HJ, Li ZR, Zhang H, Yang WJ, Ni B, et al. Gastric cancer-associated enhancement of von Willebrand factor is regulated by vascular endothelial growth factor and related to disease severity. BMC Cancer. 2015;15:80.

    Article  PubMed  PubMed Central  Google Scholar 

  25. Ishak K, Baptista A, Bianchi L, Callea F, De Groote J, Gudat F, et al. Histological grading and staging of chronic hepatitis. J Hepatol. 1995;22(6):696–9.

    Article  CAS  PubMed  Google Scholar 

  26. Ogawa K, Ohtsuki K, Shibata T, Aoki M, Nakayama M, Kitamura Y, et al. Development and evaluation of a novel (99m)tc-labeled annexin A5 for early detection of response to chemotherapy. PLoS One. 2013;8(12):e81191.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Szanto T, Vanhoorelbeke K, Toth G, Vandenbulcke A, Toth J, Noppe W, et al. Identification of a VWF peptide antagonist that blocks platelet adhesion under high shear conditions by selectively inhibiting the VWF-collagen interaction. J Thromb Haemost. 2009;7(10):1680–7.

    Article  CAS  PubMed  Google Scholar 

  28. Birkenkamp KU, Tuyt LM, Lummen C, Wierenga AT, Kruijer W, Vellenga E. The p38 MAP kinase inhibitor SB203580 enhances nuclear factor-kappa B transcriptional activity by a non-specific effect upon the ERK pathway. Br J Pharmacol. 2000;131(1):99–107.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. El Taghdouini A, Sorensen AL, Reiner AH, Coll M, Verhulst S, Mannaerts I, et al. Genome-wide analysis of DNA methylation and gene expression patterns in purified, uncultured human liver cells and activated hepatic stellate cells. Oncotarget. 2015;6(29):26729–45.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Mohar I, Brempelis KJ, Murray SA, Ebrahimkhani MR, Crispe IN. Isolation of non-parenchymal cells from the mouse liver. Methods Mol Biol. 2015;1325:3–17.

    Article  CAS  PubMed  Google Scholar 

  31. Huang X, Shao L, Gong Y, Mao Y, Liu C, Qu H, et al. A metabonomic characterization of CCl4-induced acute liver failure using partial least square regression based on the GC/MS metabolic profiles of plasma in mice. J Chromatogr B Analyt Technol Biomed Life Sci. 2008;870(2):178–85.

    Article  CAS  PubMed  Google Scholar 

  32. Mark AL, Sun Z, Warren DS, Lonze BE, Knabel MK, Melville Williams GM, et al. Stem cell mobilization is life saving in an animal model of acute liver failure. Ann Surg. 2010;252(4):591–6.

    PubMed  PubMed Central  Google Scholar 

  33. Englund GD, Bodnar RJ, Li Z, Ruggeri ZM, Du X. Regulation of von Willebrand factor binding to the platelet glycoprotein Ib-IX by a membrane skeleton-dependent inside-out signal. J Biol Chem. 2001;276(20):16952–9.

    Article  CAS  PubMed  Google Scholar 

  34. Clemetson KJ. A short history of platelet glycoprotein Ib complex. Thromb Haemost. 2007;98(1):63–8.

    CAS  PubMed  Google Scholar 

  35. Jeong JM, Kim JW, Kim DH, Park CI. First molecular cloning and gene expression analysis of teleost CD42 (glycoprotein Ib beta chain) GPIb-IX-V subunit from rock bream. Dev Comp Immunol. 2015;49(2):298–302.

    Article  CAS  PubMed  Google Scholar 

  36. Yap CL, Anderson KE, Hughan SC, Dopheide SM, Salem HH, Jackson SP. Essential role for phosphoinositide 3-kinase in shear-dependent signaling between platelet glycoprotein Ib/V/IX and integrin alpha(IIb)beta(3). Blood. 2002;99(1):151–8.

    Article  CAS  PubMed  Google Scholar 

  37. Yin H, Liu J, Li Z, Berndt MC, Lowell CA, Du X. Src family tyrosine kinase Lyn mediates VWF/GPIb-IX-induced platelet activation via the cGMP signaling pathway. Blood. 2008;112(4):1139–46.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Siller-Matula JM, Merhi Y, Tanguay JF, Duerschmied D, Wagner DD, McGinness KE, et al. ARC15105 is a potent antagonist of von Willebrand factor mediated platelet activation and adhesion. Arterioscler Thromb Vasc Biol. 2012;32(4):902–9.

    Article  CAS  PubMed  Google Scholar 

  39. McElroy AK, Erickson BR, Flietstra TD, Rollin PE, Towner JS, Nichol ST, et al. Von Willebrand factor is elevated in individuals infected with Sudan virus and is associated with adverse clinical outcomes. Viral Immunol. 2015;28(1):71–3.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Liu Y, Wang X, Li S, Hu H, Zhang D, Hu P, et al. The role of von Willebrand factor as a biomarker of tumor development in hepatitis B virus-associated human hepatocellular carcinoma: a quantitative proteomic based study. J Proteome. 2014;106:99–112.

    Article  CAS  Google Scholar 

  41. Khan MR, Ahmed D. Protective effects of Digera muricata (L.) Mart. on testis against oxidative stress of carbon tetrachloride in rat. Food Chem Toxicol. 2009;47(6):1393–9.

    Article  CAS  PubMed  Google Scholar 

  42. Ryu J, Lee JW. TM4SF5-mediated roles in the development of fibrotic phenotypes. Mediat Inflamm. 2017;2017:5108525.

    Article  Google Scholar 

  43. Li X, Wang L, Chen C. Effects of exogenous thymosin beta4 on carbon tetrachloride-induced liver injury and fibrosis. Sci Rep. 2017;7(1):5872.

    Article  PubMed  PubMed Central  Google Scholar 

  44. Rahmouni F, Saoudi M, Amri N, El-Feki A, Rebai T, Badraoui R. Protective effect of Teucrium polium on carbon tetrachloride induced genotoxicity and oxidative stress in rats. Arch Physiol Biochem. 2017:1–9.

  45. Sikka SC, Rajasekaran M, Hellstrom WJ. Role of oxidative stress and antioxidants in male infertility. J Androl. 1995;16(6):464–8.

    CAS  PubMed  Google Scholar 

  46. Minamino T, Ito Y, Ohkubo H, Shimuzu Y, Kojo K, Nishizwa N, et al. Adhesion of platelets through thromboxane A(2) receptor signaling facilitates liver repair during acute chemical-induced hepatotoxicity. Life Sci. 2015;132:85–92.

    Article  CAS  PubMed  Google Scholar 

  47. De Meyer SF, Stoll G, Wagner DD, Kleinschnitz C. von Willebrand factor: an emerging target in stroke therapy. Stroke. 2012;43(2):599–606.

    Article  PubMed  Google Scholar 

  48. Pepin M, Kleinjan A, Hajage D, Buller HR, Di Nisio M, Kamphuisen PW, et al. ADAMTS-13 and von Willebrand factor predict venous thromboembolism in patients with cancer. J Thromb Haemost. 2016;14(2):306–15.

    Article  CAS  PubMed  Google Scholar 

  49. Wu H, Yan S, Wang G, Cui S, Zhang C, Zhu Q. von Willebrand factor as a novel noninvasive predictor of portal hypertension and esophageal varices in hepatitis B patients with cirrhosis. Scand J Gastroenterol. 2015;50(9):1160–9.

    Article  CAS  PubMed  Google Scholar 

  50. Baruch Y, Neubauer K, Ritzel A, Wilfling T, Lorf T, Ramadori G. Von Willebrand gene expression in damaged human liver. Hepato-Gastroenterology. 2004;51(57):684–8.

    CAS  PubMed  Google Scholar 

  51. Fang KF, Chen ZJ, Liu M, Wu PS, Yu DZ. Blood pH in coronary artery microthrombosis of rats. Asian Pac J Trop Med. 2015;8(10):864–9.

    Article  CAS  PubMed  Google Scholar 

  52. Marsden PA, Ning Q, Fung LS, Luo X, Chen Y, Mendicino M, et al. The Fgl2/fibroleukin prothrombinase contributes to immunologically mediated thrombosis in experimental and human viral hepatitis. J Clin Invest. 2003;112(1):58–66.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Ganai AA, Khan AA, Malik ZA, Farooqi H. Genistein modulates the expression of NF-kappaB and MAPK (p-38 and ERK1/2), thereby attenuating d-Galactosamine induced fulminant hepatic failure in Wistar rats. Toxicol Appl Pharmacol. 2015;283(2):139–46.

    Article  CAS  PubMed  Google Scholar 

  54. Ma L, Gong X, Kuang G, Jiang R, Chen R, Wan J. Sesamin ameliorates lipopolysaccharide/d-galactosamine-induced fulminant hepatic failure by suppression of Toll-like receptor 4 signaling in mice. Biochem Biophys Res Commun. 2015;461(2):230–6.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We would like to thank Chen-hui Wang for isolating the LSECs from mice and Xiao-he Shen for technical assistance in the Ets-1 siRNA transduction investigations.

Funding

This research was supported by grants from the National Natural Science Foundation of China (no. 31500710 and no. 31500720) and the National Key Research and Development Project (2016YFA0502204).

Author information

Author notes

  1. Hai-Jian Sun, Jian Chen, and Hao Zhang contributed equally to this study.

Authors and Affiliations

  1. Institute of Immunology, Third Military Medical University (Military Medical University of the Army), PLA, Chongqing, 400038, People’s Republic of China

    Hai-Jian Sun, Jian Chen, Hao Zhang, Yu-Zhang Wu & Xia Yang

  2. Embryology, Nanjing Medical University, Nanjing, Jiangsu, 210029, People’s Republic of China

    Hai-Jian Sun

  3. Out-patient Department of NO. 75737 Army, PLA, Guangzhou, 510800, People’s Republic of China

    Hao Zhang

  4. Department of Pathophysiology and High Altitude Pathology, Third Military Medical University (Military Medical University of the Army), Chongqing, 400038, People’s Republic of China

    Bing Ni

  5. van Velkinburgh Initiative for Collaboratory BioMedical Research, Santa Fe, NM, 87501, USA

    Jennifer C. van Velkinburgh

  6. Department of Pharmacy, Southwestern Hospital, Chongqing, 400038, People’s Republic of China

    Yao Liu

Authors
  1. Hai-Jian Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jian Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hao Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Bing Ni
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jennifer C. van Velkinburgh
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yao Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yu-Zhang Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xia Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

XY and YZW conceived of and designed the study; HJS, JC, and HZ carried out the experiments; YL collected the specimens from patients and healthy donors; XY, HJS, JC, HZ, and BN analyzed and interpreted the data; JCvV made critical revision to the article for important intellectual content. All authors made substantial contributions towards drafting the manuscript, reviewing the final manuscript for intellectual content, and authorizing the submission.

Corresponding authors

Correspondence to Yu-Zhang Wu or Xia Yang.

Ethics declarations

Conflict of interests

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sun, HJ., Chen, J., Zhang, H. et al. Von Willebrand factor protects against acute CCl4-induced hepatotoxicity through phospho-p38 MAPK signaling pathway inhibition. Immunol Res 65, 1046–1058 (2017). https://doi.org/10.1007/s12026-017-8946-7

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12026-017-8946-7

Keywords

Search

Navigation