Advertisement

Mesenchymal stem cells suppress hepatic fibrosis accompanied by expanded intrahepatic natural killer cells in rat fibrosis model

  • Deniz Guney DumanEmail author
  • Noushin Zibandeh
  • Mustafa Umit Ugurlu
  • Cigdem Celikel
  • Tolga Akkoc
  • Munkhtsetseg Banzragch
  • Deniz Genc
  • Osman Ozdogan
  • Tunç Akkoc
Original Article

Abstract

Natural killer (NK) cells have antifibrotic effects. We have evaluated the influence of rat bone marrow-mesenchymal stem cell (BM-MSC) treatment on liver histology, biochemical liver function tests, systemic immunoregulatory state and NK cell distribution in liver and peripheral blood in rat model of common bile duct (CBD) ligation and compared the results with the control group. Rats were divided into three groups: (1) CBD ligated (CBDL) rats received phosphate-buffered saline (CBDL + PBS group) or (2) MSC (CBDL + MSC group) and sham-operated rats received MSC (healthy + MSC group). We found significantly decreased fibrosis scores with BM-MSC treatment in CBDL rats compared to the control (CBDL + PBS) group while no fibrosis developed in sham operated (healthy + MSC) group. BM-MSC treatment has decreased the inflammation as reflected by the significantly decreased T cell proliferation and inflammatory cytokine concentrations from splenocyte culture and liver tissue itself compared to CBDL + PBS. NK cells both in parenchyme and portal areas decreased significantly in liver and blood in CBDL + PBS compared to healthy + MSC while they were found to be increased in CBDL + MSC compared to CBDL + PBS rats. In conclusion, BM-MSCs may suppress hepatic fibrosis accompanied by expanded intrahepatic NK cells in CBDL rats. Thus, our animal study shows that MSC treatment holds great promise for treatment of patients with end-stage liver diseases through a possible mechanism by adopting the NK cell population and new studies investigating the role of NK cells and clinical fibrosis are warranted.

Trial registration number: Marmara University Animal Care and Use Committee approval code: 73.2013.mar.

Keywords

Cell therapy Stem cells Animal models Natural killer cells 

Abbreviations

NK cells

Natural killer cells

MSC

Mesenchymal stem cells

BM

Bone marrow

CBD

Common bile duct

CBDL

Common bile duct ligated

IL

Interleukin

TNF

Tumor necrosis factor

IFN-γ

Interferon-γ

ABMI

Autologous bone marrow cell infusion

HSCs

Hepatic stellate cells

AST

Aspartate aminotransferase

ALT

Alanine aminotransferase

BUN

Blood urea nitrogen

PBS

Phosphate buffer saline

DMEM

Dulbecco’s modified Eagle medium

FBS

Fetal bovine serum

GFP

Green fluorescent protein

CFSE

Carboxyfluorescein succinimidyl ester

HGF

Hepatocytes growth factor

MMPs

Matrix metalloproteinases

SOCS1

Cytokine signaling 1

Treg cells

T regulatory cells

Notes

Acknowledgements

This study was funded by the grant (SAG-A-041213-0443) from Marmara University Scientific Research Project Commission.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest regarding the publication of this article.

Supplementary material

11033_2019_4736_MOESM1_ESM.tif (53 kb)
Supplementary material Scheme of study protocol. Fibrosis was induced by common bile duct ligation which was taken as time zero. On day 14, MSCs (1 × 106) were injected through tail vein. Rats were sacrificed on day 28 (TIF 53 KB)

References

  1. 1.
    Schuppan D, Afdhal NH (2008) Liver cirrhosis. Lancet 371:838–851CrossRefGoogle Scholar
  2. 2.
    Jang YO, Jun BG, Baik SK, Kim MY, Kwon SO (2015) Inhibition of hepatic stellate cells by bone marrow-derived mesenchymal stem cells in hepatic fibrosis. Clin Mol Hepatol 21:141–149CrossRefGoogle Scholar
  3. 3.
    Terai S, Ishikawa T, Omori K et al (2006) Improved liver function in patients with liver cirrhosis after autologous bone marrow cell infusion therapy. Stem cells 24:2292–2298CrossRefGoogle Scholar
  4. 4.
    Gaia S, Smedile A, Omede P et al (2006) Feasibility and safety of G-CSF administration to induce bone marrow-derived cells mobilization in patients with end stage liver disease. J Hepatol 45:13–19CrossRefGoogle Scholar
  5. 5.
    Kim JK, Kim SJ, Kim Y et al (2017) Long-term follow-up of patients after autologous bone marrow cell infusion for decompensated liver cirrhosis. Cell Transplant 26:1059–1066CrossRefGoogle Scholar
  6. 6.
    Jeong WI, Park O, Suh YG et al (2011) Suppression of innate immunity (natural killer cell/interferon γ) in the advanced stages of liver fibrosis in mice. Hepatology 53:1342–1351CrossRefGoogle Scholar
  7. 7.
    Muhanna N, Tair LA, Doron S et al (2011) Amelioration of hepatic fibrosis by NK cell activation. Gut 60:90–98CrossRefGoogle Scholar
  8. 8.
    Melhem A, Muhanna N, Bishara A et al (2006) Anti-fibrotic activity of NK cells in experimental liver injury through killing of activated HSC. J Hepatol 45:60–71CrossRefGoogle Scholar
  9. 9.
    Seki E, Schwabe RF (2015) Hepatic inflammation and fibrosis: functional links and key pathways. Hepatology 61(3):1066–1079.  https://doi.org/10.1002/hep.27332 CrossRefGoogle Scholar
  10. 10.
    Li X, Zhang M, Liu J et al (2016) Intrahepatic NK cells function suppressed in advanced liver fibrosis. Eur J Clin Invest 46:864–872CrossRefGoogle Scholar
  11. 11.
    Liedtke C, Luedde T, Sauerbruch T et al (2013) Experimental liver fibrosis research: update on animal models, legal issues and translational aspects. Fibrogenesis Tissue Repair 6:19CrossRefGoogle Scholar
  12. 12.
    Geerts AM, Vanheule E, Praet M, Van Vlierberghe H, De Vos M, Colle I (2008) Comparison of three research models of portal hypertension in mice: macroscopic, histological and portal pressure evaluation. Int J Exp Pathol 89:251–263CrossRefGoogle Scholar
  13. 13.
    Ishak K, Baptista A, Bianchi L et al (1995) Histological grading and staging of chronic hepatitis. J Hepatology 22:696–699CrossRefGoogle Scholar
  14. 14.
    Hirazawa K, Hazama S, Oka M (1999) Depressed cytotoxic activity of hepatic nonparenchymal cells in rats with obstructive jaundice. Surgery 126:900–907CrossRefGoogle Scholar
  15. 15.
    Koyama Y, Brenner DA (2017) Liver inflammation and fibrosis. J Clin Invest 3(1):55–64 127)CrossRefGoogle Scholar
  16. 16.
    Li Q, Zhou X, Shi Y et al (2013) In vivo tracking and comparison of the therapeutic effects of MSCs and HSCs for liver injury. PLoS One 8:e62363CrossRefGoogle Scholar
  17. 17.
    Quintanilha LF, Mannheimer EG, Carvalho AB et al (2008) Bone marrow cell transplant does not prevent or reverse murine liver cirrhosis. Cell Transpl 17(8):943–953CrossRefGoogle Scholar
  18. 18.
    Ahmed SK, Mohammed SA, Khalaf G, Fikry H (2014) Role of bone marrow mesenchymal stem cells in the treatment of CCL4 induced liver fibrosis in albino rats: a histological and immunohistochemical Study. Int J Stem Cells 7(2):87–97CrossRefGoogle Scholar
  19. 19.
    Jeong WI, Park O, Suh YG et al (2011) Suppression of immunity (natural killer cell/interferon γ) in the advanced stages of liver fibrosis in mice. Hepatology 53:1342–1351CrossRefGoogle Scholar
  20. 20.
    Fang XQ, Zhang JF, Song HY et al (2016) Effect of umbilical cord mesenchymal stem cell transplantation on immune function and prognosis of patients with decompensated hepatitis B cirrhosis. Zhonghua Gan Zang Bing Za Zhi 24:907–910Google Scholar
  21. 21.
    Zhao L, Chen S, Shi X, Cao H, Li L (2018) A pooled analysis of mesenchymal stem cell-based therapy for liver disease. Stem Cell Res Ther 9:72CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Gastroenterology, School of MedicineMarmara UniversityIstanbulTurkey
  2. 2.Department of Pediatric Allergy-Immunology, School of MedicineMarmara UniversityIstanbulTurkey
  3. 3.Department of General Surgery, School of MedicineMarmara UniversityIstanbulTurkey
  4. 4.Department of Pathology, School of MedicineMarmara UniversityIstanbulTurkey
  5. 5.Genetic Engineering and Biotechnology InstituteTubitak Marmara Research CenterKocaeliTurkey
  6. 6.Department of GastroenterologyDarica Farabi State HospitalKocaeliTurkey
  7. 7.Department of GastroenterologyMarmara University Pendik Education and Research HospitalIstanbulTurkey

Personalised recommendations