Skip to main content

Advertisement

SpringerLink
Log in

Role of cell autophagy in the generation of IgM and hepatic fibrosis in primary biliary cholangitis

  • Original Article
  • Published:
Clinical Rheumatology Aims and scope Submit manuscript

Abstract

Objective

Autophagy in the immune and autoimmune diseases has been concerned more and more. We investigated the potential role of cell autophagy of B cells generating IgM in primary biliary cholangitis (PBC), and explored the relationship between the cell autophagy of hepatic stellate cells (HSCs) and hepatic fibrosis in PBC.

Methods

We examined the aggregation degree of microtubule-associated protein light chain 3II (LC3II) in B cells of PBC (n = 21), health control (HC, n = 15) and disease control (DC, n = 8, active hepatitis B). The expression level of p62/SQSTM1 (p62) in B cells was detected by flow cytometry. ELISA was adopted to detect the level of IgM in the B cell culture supernatant under the basal condition, activated condition(stimulated by pokeweed mitogen, PWM) and inhibited condition(inhibited by autophagy inhibitor, 3-methyladenine, 3-MA) respectively. We detected the expression of α-smooth muscle actin (α-SMA), the infiltration level of LC3II, transforming growth factor-β (TGF-β), platelet-derived growth factor-bb (PDGF-bb), and collagen I in the hepatic tissues of five PBC patients and two HC individuals.

Results

When B cells were stimulated and activated by PWM, the expression of p62 increased, and the mean fluorescence intensity (MFI, 2404.8 ± 689.0) of p62 in PBC was lower than that in HC (2966.8 ± 488.9,P = 0.0227). Under 3-MA treatment, the MFI expressed of p62 in B cells weakened. The reduction difference in PBC (466.4 ± 214.9) was smaller than that in HC (1166.6 ± 231.2, P = 0.0000) and DC (1184.8 ± 197.7, P = 0.0001). The level of generating IgM in the PBC group was obviously higher than that in the HC group (65.7 ± 15.4 vs. 49.5 ± 20.4 ng/ml, P = 0.0357). Before and after B cells were treated with 3-MA, the peak level of IgM did not significantly diminish in PBC and DC (65.7 ± 15.4 vs. 59.6 ± 18.7 ng/ml, P = 0.2965; 50.1 ± 17.4 vs. 48.4 ± 12.3 ng/ml, P = 0.8336). The immunohistochemical result revealed that the expression level of collagen I, α-SMA, TGF-β and PDGF-bb in PBC was higher than that in HC, but the expression of LC3II was lower. Similarly, immunofluorescence assay revealed that the fluorescence intensity of collagen I was higher but LC3II was lower in PBC than that in HC.

Conclusion

The high level autophagy of B cells from PBC patients is one important factor to synthesize and secrete IgM. Hepatic fibrosis of PBC is probably associated with the weakened autophagy of HSCs.

Key Points

• High-level autophagy of B cells in PBC patients is an important factor to synthesize and secrete IgM.

• Pro-fibrogenic cytokines in PBC influence the increase of collagen-I through HSC autophagy, and promote the occurrence and development of hepatic fibrosis.

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

Similar content being viewed by others

References

  1. Beuers U, Gershwin ME, Gish RG, Invernizzi P, Jones DEJ, Lindor K, Ma X, Mackay IR, Parés A, Tanaka A, Vierling JM, Poupon R (2015) Changing nomenclature for PBC: from ‘cirrhosis’ to ‘cholangitis’. Hepatology. 62(5):1620–1622

    Article  Google Scholar 

  2. Carey EJ, Ali AH, Lindor KD (2015) Primary biliary cirrhosis. Lancet 386(10003):1565–1575

    Article  CAS  Google Scholar 

  3. Pengo N, Scolari M, Oliva L, Milan E, Mainoldi F, Raimondi A, Fagioli C, Merlini A, Mariani E, Pasqualetto E, Orfanelli U, Ponzoni M, Sitia R, Casola S, Cenci S (2013) Plasma cells require autophagy for sustainable immunoglobulin production. Nat Immunol 14(3):298–305

    Article  CAS  Google Scholar 

  4. Kimura S, Fujita N, Noda T et al (2009) Monitoring autophagy in mammalian cultured cells through the dynamics of LC3. Methods Enzymol 452:1–12

    Article  CAS  Google Scholar 

  5. Popov Y, Schuppan D (2009) Targeting liver fibrosis: strategies for development and validation of antifibrotic therapies. Hepatology. 50(4):1294–1306

    Article  CAS  Google Scholar 

  6. DeLeve LD (2015) Liver sinusoidal endothelial cells in hepatic fibrosis. Hepatology. 61(5):1740–1746

    Article  CAS  Google Scholar 

  7. Hidvegi T, Ewing M, Hale P, Dippold C, Beckett C, Kemp C, Maurice N, Mukherjee A, Goldbach C, Watkins S, Michalopoulos G, Perlmutter DH (2010) An autophagy-enhancing drug promotes degradation of mutant alpha1-antitrypsin Z and reduces hepatic fibrosis. Science 329:229–232

    Article  CAS  Google Scholar 

  8. Thoen LF, Guimaraes EL, Dolle L et al (2011) A role for autophagy during hepatic stellate cell activation. J Hepatol 55(6):1353–1360

    Article  CAS  Google Scholar 

  9. Shuai Z, Wang J, Badamagunta M et al (2017) The fingerprint of antimitochondrial antibodies and the etiology of primary biliary cholangitis. Hepatology 65(5):1670–1682

    Article  CAS  Google Scholar 

  10. Invernizzi P, Ransom M, Raychaudhuri S et al (2012) Classical HLA-DRB1 and DPB1 alleles account for HLA associations with primary biliary cirrhosis. Genes Immun 13:461–468

    Article  CAS  Google Scholar 

  11. Tanaka A, Leung PSC, Gershwin ME (2018) The genetics and epigenetics of primary biliary cholangitis. [J]. Clin Liver Dis 22:443–455

    Article  Google Scholar 

  12. Boonstra K, Beuers U, Ponsioen CY (2012) Epidemiology of primary sclerosing cholangitis and primary biliary cirrhosis: a systematic review. J Hepatol 56(5):1181–1188

    Article  Google Scholar 

  13. Liu H, Liu Y, Wang L, Xu D, Lin B, Zhong R, Gong S, Podda M, Invernizzi P (2010) Prevalence of primary biliary cirrhosis in adults referring hospital for annual health check-up in Southern China. BMC Gastroenterol 10:100

    Article  Google Scholar 

  14. Lindor KD, Gershwin ME, Poupon R, Kaplan M, Bergasa NV, Heathcote EJ, American Association for Study of Liver Diseases (2009) Primary biliary cirrhosis. Hepatology 50:291–308

    Article  Google Scholar 

  15. Sakauchi F, Mori M, Zeniya M et al (2006) Antimitochondrial antibody negative primary biliary cirrhosis in Japan: utilization of clinical data when patients applied to receive public financial aid. J Epidemiol 16:30–34

    Article  Google Scholar 

  16. Lindgren S, Eriksson S (1982) IgM in primary biliary cirrhosis. Physicochemical and complement activating properties. J Lab Clin Med 99(5):636–645

    CAS  PubMed  Google Scholar 

  17. Hilscher M, Hernandez-Gea V, Friedman SL (2012) Autophagy and mesenchymal cell fibrogenesis. Biochim Biophys Acta 1831(7):972–978

    PubMed  Google Scholar 

  18. Kim SI, Na HJ, Ding Y, Wang Z, Lee SJ, Choi ME (2012) Autophagy promotes intracellular degradation of type I collagen induced by transforming growth factor (TGF)-beta1. J Biol Chem 287(15):11677–11688

    Article  CAS  Google Scholar 

  19. Melton AC, Yee HF (2007) Hepatic stellate cell protrusions couple platelet-derived growth factor-BB to chemotaxis. Hepatology 45(6):1446–1453

    Article  CAS  Google Scholar 

  20. Seki E, De Minicis S, Osterreicher CH et al (2007) TLR4 enhances TGF-beta signaling and hepatic fibrosis. Nat Med 13(11):1324–1332

    Article  CAS  Google Scholar 

  21. Friedman SL (2008) Mechanisms of hepatic fibrogenesis. Gastroenterology. 134(6):1655–1669

    Article  CAS  Google Scholar 

  22. Leask A, Abraham DJ (2004) TGF-beta signaling and the fibrotic response. FASEB J 18(7):816–827

    Article  CAS  Google Scholar 

  23. Xu Y, Yang S, Huang J, Ruan S, Zheng Z, Lin J (2012) Tgf-beta1 induces autophagy and promotes apoptosis in renal tubular epithelial cells. Int J Mol Med 29(5):781–790

    CAS  PubMed  Google Scholar 

  24. Harris J (2011) Autophagy and cytokines. Cytokine. 56(2):140–144

    Article  CAS  Google Scholar 

  25. M NSaT (2012) The role of TGF-β receptors in firosis. Open Rheumatol 6:156–162

    Article  Google Scholar 

  26. Suzuki HI, Kiyono K, Miyazono K (2010) Regulation of autophagyby transforming growth factor-β (TGF-β) signaling. Autophagy. 6:645–647

    Article  Google Scholar 

  27. Wang RC, Levine B (2010) Autophagy in cellular growth control. FEBS Lett 584(7):1417–1426

    Article  CAS  Google Scholar 

  28. Pinzani MMS, Grappone C et al (1994) Expression of platelet-derived growth factor in a model of acute liver injury. Hepatology 19:701–707

    Article  CAS  Google Scholar 

  29. Borkham-Kamphorst E, van Roeyen CR, Ostendorf T et al (2007) Pro-fibrogenic potential of PDGF-D in liver fibrosis. J Hepatol 46(6):1064–1074

    Article  CAS  Google Scholar 

  30. Kazlauskas A (2017) PDGFs and their receptors. Gene. 614:1–7

    Article  CAS  Google Scholar 

  31. Salabei JK, Cummins TD, Singh M, Jones SP, Bhatnagar A, Hill BG (2013) PDGF-mediated autophagy regulates vascular smooth muscle cell phenotype and resistance to oxidative stress. Biochem J 451(3):375–388

    Article  CAS  Google Scholar 

  32. Bataller R, Brenner DA (2005) Liver fibrosis. J Clin Investig 115(2):209–218

    Article  CAS  Google Scholar 

  33. Hernandez-Gea V, Ghiassi-Nejad Z, Rozenfeld R et al (2012) Autophagy releases lipid that promotes fibrogenesis by activated hepatic stellate cells in mice and in human tissues. Gastroenterology 142(4):938–946

    Article  Google Scholar 

Download references

Funding

This work was supported by The National Natural Science Foundation of China [No.81571594, No.81771764].

Author information

Authors and Affiliations

  1. Department of Rheumatology, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Key Laboratory of Rheumatology and Clinical Immunology, Ministry of Education, No. 1 Shuai Fu Yuan, Eastern District, Beijing, 100730, China

    Chaofeng Lian, Yun Zhao, Jinlei Sun, Liling Zhao & Fengchun Zhang

Authors
  1. Chaofeng Lian
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yun Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jinlei Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Liling Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Fengchun Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Chaofeng Lian drafted the manuscript, Chaofeng Lian and Yun Zhao designed and analyzed data, Jinlei Sun and Liling Zhao revised the manuscript, Fengchun Zhang designed the study and drafted and revised the manuscript. All authors approved submission of the manuscript. We agree to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.

Corresponding author

Correspondence to Fengchun Zhang.

Ethics declarations

Disclosures

None.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lian, C., Zhao, Y., Sun, J. et al. Role of cell autophagy in the generation of IgM and hepatic fibrosis in primary biliary cholangitis. Clin Rheumatol 39, 3499–3506 (2020). https://doi.org/10.1007/s10067-020-05111-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10067-020-05111-6

Keywords

Search

Navigation