Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1998 May 1;101(9):1960–1969. doi: 10.1172/JCI504

Concanavalin A-induced liver injury triggers hepatocyte proliferation.

C Trautwein 1, T Rakemann 1, N P Malek 1, J Plümpe 1, G Tiegs 1, M P Manns 1
PMCID: PMC508783  PMID: 9576761

Abstract

Concanavalin A (Con A) injection into mice leads to immune-mediated liver injury. We studied whether after Con A-induced liver injury, TNF- and IL-6-dependent signaling pathways known to be related to hepatocyte proliferation are activated. 2 h after Con A injection, maximum TNF-alpha, and after 4-8 h, maximum IL-6 serum levels were found. The rise in aminotransferases and DNA fragmentation started after 4 h; maximum levels were evident after 8 h. 5-Bromo-2'-deoxyuridine staining and nuclear cyclin A expression as markers of the S-phase were first detected in hepatocyte nuclei after 24 h, peaking after 48 h. An increase in TNF-dependent nuclear expression of CCAAT/enhancer-binding protein-beta (C/EBP-beta)/liver-enriched activating protein (LAP) was detected after 1 h, whereas an increase in RNA expression was evident only after 4 h. C/EBP-beta/LAP expression returned to normal values before progression into the S-phase. DNA binding of signal transducer and activator of transcription (STAT) 3/acute phase response factor (APRF) increased for up to 8 h. As found by supershift experiments, in addition to STAT3/APRF, STAT1 also binds to the same sequence. During the course of time gel shift experiments, DNA binding of the apoptosis-related STAT1 started earlier than DNA binding of STAT3/APRF, which regulates hepatocyte proliferation. However, the subsequent decrease in DNA binding of both factors was comparable. This study demonstrates that after Con A injection, TNF- and IL-6- dependent signals trigger nuclear events regulating hepatocyte apoptosis and proliferation during liver injury.

Full Text

The Full Text of this article is available as a PDF (633.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akerman P., Cote P., Yang S. Q., McClain C., Nelson S., Bagby G. J., Diehl A. M. Antibodies to tumor necrosis factor-alpha inhibit liver regeneration after partial hepatectomy. Am J Physiol. 1992 Oct;263(4 Pt 1):G579–G585. doi: 10.1152/ajpgi.1992.263.4.G579. [DOI] [PubMed] [Google Scholar]
  2. Akira S., Nishio Y., Inoue M., Wang X. J., Wei S., Matsusaka T., Yoshida K., Sudo T., Naruto M., Kishimoto T. Molecular cloning of APRF, a novel IFN-stimulated gene factor 3 p91-related transcription factor involved in the gp130-mediated signaling pathway. Cell. 1994 Apr 8;77(1):63–71. doi: 10.1016/0092-8674(94)90235-6. [DOI] [PubMed] [Google Scholar]
  3. Barone M. V., Crozat A., Tabaee A., Philipson L., Ron D. CHOP (GADD153) and its oncogenic variant, TLS-CHOP, have opposing effects on the induction of G1/S arrest. Genes Dev. 1994 Feb 15;8(4):453–464. doi: 10.1101/gad.8.4.453. [DOI] [PubMed] [Google Scholar]
  4. Bock C. T., Tillmann H. L., Maschek H. J., Manns M. P., Trautwein C. A preS mutation isolated from a patient with chronic hepatitis B infection leads to virus retention and misassembly. Gastroenterology. 1997 Dec;113(6):1976–1982. doi: 10.1016/s0016-5085(97)70018-0. [DOI] [PubMed] [Google Scholar]
  5. Bromberg J. F., Horvath C. M., Wen Z., Schreiber R. D., Darnell J. E., Jr Transcriptionally active Stat1 is required for the antiproliferative effects of both interferon alpha and interferon gamma. Proc Natl Acad Sci U S A. 1996 Jul 23;93(15):7673–7678. doi: 10.1073/pnas.93.15.7673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Buck M., Turler H., Chojkier M. LAP (NF-IL-6), a tissue-specific transcriptional activator, is an inhibitor of hepatoma cell proliferation. EMBO J. 1994 Feb 15;13(4):851–860. doi: 10.1002/j.1460-2075.1994.tb06328.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cressman D. E., Diamond R. H., Taub R. Rapid activation of the Stat3 transcription complex in liver regeneration. Hepatology. 1995 May;21(5):1443–1449. [PubMed] [Google Scholar]
  8. Cressman D. E., Greenbaum L. E., DeAngelis R. A., Ciliberto G., Furth E. E., Poli V., Taub R. Liver failure and defective hepatocyte regeneration in interleukin-6-deficient mice. Science. 1996 Nov 22;274(5291):1379–1383. doi: 10.1126/science.274.5291.1379. [DOI] [PubMed] [Google Scholar]
  9. Darnell J. E., Jr, Kerr I. M., Stark G. R. Jak-STAT pathways and transcriptional activation in response to IFNs and other extracellular signaling proteins. Science. 1994 Jun 3;264(5164):1415–1421. doi: 10.1126/science.8197455. [DOI] [PubMed] [Google Scholar]
  10. Diehl A. M., Rai R. M. Liver regeneration 3: Regulation of signal transduction during liver regeneration. FASEB J. 1996 Feb;10(2):215–227. doi: 10.1096/fasebj.10.2.8641555. [DOI] [PubMed] [Google Scholar]
  11. Diehl A. M., Yang S. Q., Yin M., Lin H. Z., Nelson S., Bagby G. Tumor necrosis factor-alpha modulates CCAAT/enhancer binding proteins-DNA binding activities and promotes hepatocyte-specific gene expression during liver regeneration. Hepatology. 1995 Jul;22(1):252–261. doi: 10.1016/0270-9139(95)90379-8. [DOI] [PubMed] [Google Scholar]
  12. Diehl A. M., Yin M., Fleckenstein J., Yang S. Q., Lin H. Z., Brenner D. A., Westwick J., Bagby G., Nelson S. Tumor necrosis factor-alpha induces c-jun during the regenerative response to liver injury. Am J Physiol. 1994 Oct;267(4 Pt 1):G552–G561. doi: 10.1152/ajpgi.1994.267.4.G552. [DOI] [PubMed] [Google Scholar]
  13. Ferrari C., Mondelli M. U., Penna A., Fiaccadori F., Chisari F. V. Functional characterization of cloned intrahepatic, hepatitis B virus nucleoprotein-specific helper T cell lines. J Immunol. 1987 Jul 15;139(2):539–544. [PubMed] [Google Scholar]
  14. Fukada T., Hibi M., Yamanaka Y., Takahashi-Tezuka M., Fujitani Y., Yamaguchi T., Nakajima K., Hirano T. Two signals are necessary for cell proliferation induced by a cytokine receptor gp130: involvement of STAT3 in anti-apoptosis. Immunity. 1996 Nov;5(5):449–460. doi: 10.1016/s1074-7613(00)80501-4. [DOI] [PubMed] [Google Scholar]
  15. Gantner F., Leist M., Lohse A. W., Germann P. G., Tiegs G. Concanavalin A-induced T-cell-mediated hepatic injury in mice: the role of tumor necrosis factor. Hepatology. 1995 Jan;21(1):190–198. doi: 10.1016/0270-9139(95)90428-x. [DOI] [PubMed] [Google Scholar]
  16. Horvath C. M., Wen Z., Darnell J. E., Jr A STAT protein domain that determines DNA sequence recognition suggests a novel DNA-binding domain. Genes Dev. 1995 Apr 15;9(8):984–994. doi: 10.1101/gad.9.8.984. [DOI] [PubMed] [Google Scholar]
  17. Hsu H., Shu H. B., Pan M. G., Goeddel D. V. TRADD-TRAF2 and TRADD-FADD interactions define two distinct TNF receptor 1 signal transduction pathways. Cell. 1996 Jan 26;84(2):299–308. doi: 10.1016/s0092-8674(00)80984-8. [DOI] [PubMed] [Google Scholar]
  18. Küsters S., Gantner F., Künstle G., Tiegs G. Interferon gamma plays a critical role in T cell-dependent liver injury in mice initiated by concanavalin A. Gastroenterology. 1996 Aug;111(2):462–471. doi: 10.1053/gast.1996.v111.pm8690213. [DOI] [PubMed] [Google Scholar]
  19. Leist M., Gantner F., Bohlinger I., Germann P. G., Tiegs G., Wendel A. Murine hepatocyte apoptosis induced in vitro and in vivo by TNF-alpha requires transcriptional arrest. J Immunol. 1994 Aug 15;153(4):1778–1788. [PubMed] [Google Scholar]
  20. Leist M., Gantner F., Jilg S., Wendel A. Activation of the 55 kDa TNF receptor is necessary and sufficient for TNF-induced liver failure, hepatocyte apoptosis, and nitrite release. J Immunol. 1995 Feb 1;154(3):1307–1316. [PubMed] [Google Scholar]
  21. Liu Z. G., Hsu H., Goeddel D. V., Karin M. Dissection of TNF receptor 1 effector functions: JNK activation is not linked to apoptosis while NF-kappaB activation prevents cell death. Cell. 1996 Nov 1;87(3):565–576. doi: 10.1016/s0092-8674(00)81375-6. [DOI] [PubMed] [Google Scholar]
  22. Löhr H. F., Schlaak J. F., Lohse A. W., Böcher W. O., Arenz M., Gerken G., Meyer Zum Büschenfelde K. H. Autoreactive CD4+ LKM-specific and anticlonotypic T-cell responses in LKM-1 antibody-positive autoimmune hepatitis. Hepatology. 1996 Dec;24(6):1416–1421. doi: 10.1002/hep.510240619. [DOI] [PubMed] [Google Scholar]
  23. Löhr H. F., Weber W., Schlaak J., Goergen B., Meyer zum Buschenfelde K. H., Gerken G. Proliferative response of CD4+ T cells and hepatitis B virus clearance in chronic hepatitis with or without hepatitis B e-minus hepatitis B virus mutants. Hepatology. 1995 Jul;22(1):61–68. doi: 10.1002/hep.1840220110. [DOI] [PubMed] [Google Scholar]
  24. Mizuhara H., O'Neill E., Seki N., Ogawa T., Kusunoki C., Otsuka K., Satoh S., Niwa M., Senoh H., Fujiwara H. T cell activation-associated hepatic injury: mediation by tumor necrosis factors and protection by interleukin 6. J Exp Med. 1994 May 1;179(5):1529–1537. doi: 10.1084/jem.179.5.1529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Napoli J., Bishop G. A., McGuinness P. H., Painter D. M., McCaughan G. W. Progressive liver injury in chronic hepatitis C infection correlates with increased intrahepatic expression of Th1-associated cytokines. Hepatology. 1996 Oct;24(4):759–765. doi: 10.1002/hep.510240402. [DOI] [PubMed] [Google Scholar]
  26. Sherr C. J. Mammalian G1 cyclins. Cell. 1993 Jun 18;73(6):1059–1065. doi: 10.1016/0092-8674(93)90636-5. [DOI] [PubMed] [Google Scholar]
  27. Tiegs G., Hentschel J., Wendel A. A T cell-dependent experimental liver injury in mice inducible by concanavalin A. J Clin Invest. 1992 Jul;90(1):196–203. doi: 10.1172/JCI115836. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Trautwein C., Caelles C., van der Geer P., Hunter T., Karin M., Chojkier M. Transactivation by NF-IL6/LAP is enhanced by phosphorylation of its activation domain. Nature. 1993 Aug 5;364(6437):544–547. doi: 10.1038/364544a0. [DOI] [PubMed] [Google Scholar]
  29. Trautwein C., Rakemann T., Niehof M., Rose-John S., Manns M. P. Acute-phase response factor, increased binding, and target gene transcription during liver regeneration. Gastroenterology. 1996 Jun;110(6):1854–1862. doi: 10.1053/gast.1996.v110.pm8964411. [DOI] [PubMed] [Google Scholar]
  30. Trautwein C., Rakemann T., Pietrangelo A., Plümpe J., Montosi G., Manns M. P. C/EBP-beta/LAP controls down-regulation of albumin gene transcription during liver regeneration. J Biol Chem. 1996 Sep 6;271(36):22262–22270. doi: 10.1074/jbc.271.36.22262. [DOI] [PubMed] [Google Scholar]
  31. Trautwein C., Walker D. L., Plümpe J., Manns M. P. Transactivation of LAP/NF-IL6 is mediated by an acidic domain in the N-terminal part of the protein. J Biol Chem. 1995 Jun 23;270(25):15130–15136. doi: 10.1074/jbc.270.25.15130. [DOI] [PubMed] [Google Scholar]
  32. Wegner M., Cao Z., Rosenfeld M. G. Calcium-regulated phosphorylation within the leucine zipper of C/EBP beta. Science. 1992 Apr 17;256(5055):370–373. doi: 10.1126/science.256.5055.370. [DOI] [PubMed] [Google Scholar]
  33. Wen Z., Zhong Z., Darnell J. E., Jr Maximal activation of transcription by Stat1 and Stat3 requires both tyrosine and serine phosphorylation. Cell. 1995 Jul 28;82(2):241–250. doi: 10.1016/0092-8674(95)90311-9. [DOI] [PubMed] [Google Scholar]
  34. Williams S. C., Baer M., Dillner A. J., Johnson P. F. CRP2 (C/EBP beta) contains a bipartite regulatory domain that controls transcriptional activation, DNA binding and cell specificity. EMBO J. 1995 Jul 3;14(13):3170–3183. doi: 10.1002/j.1460-2075.1995.tb07319.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yamada Y., Kirillova I., Peschon J. J., Fausto N. Initiation of liver growth by tumor necrosis factor: deficient liver regeneration in mice lacking type I tumor necrosis factor receptor. Proc Natl Acad Sci U S A. 1997 Feb 18;94(4):1441–1446. doi: 10.1073/pnas.94.4.1441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yin M., Yang S. Q., Lin H. Z., Lane M. D., Chatterjee S., Diehl A. M. Tumor necrosis factor alpha promotes nuclear localization of cytokine-inducible CCAAT/enhancer binding protein isoforms in hepatocytes. J Biol Chem. 1996 Jul 26;271(30):17974–17978. doi: 10.1074/jbc.271.30.17974. [DOI] [PubMed] [Google Scholar]
  37. Zhong Z., Wen Z., Darnell J. E., Jr Stat3: a STAT family member activated by tyrosine phosphorylation in response to epidermal growth factor and interleukin-6. Science. 1994 Apr 1;264(5155):95–98. doi: 10.1126/science.8140422. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES