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. 1998 Aug 1;102(3):538–549. doi: 10.1172/JCI1018

Mechanisms of spontaneous resolution of rat liver fibrosis. Hepatic stellate cell apoptosis and reduced hepatic expression of metalloproteinase inhibitors.

J P Iredale 1, R C Benyon 1, J Pickering 1, M McCullen 1, M Northrop 1, S Pawley 1, C Hovell 1, M J Arthur 1
PMCID: PMC508915  PMID: 9691091

Abstract

Liver fibrosis results from the excessive secretion of matrix proteins by hepatic stellate cells (HSC), which proliferate during fibrotic liver injury. We have studied a model of spontaneous recovery from liver fibrosis to determine the biological mechanisms mediating resolution. Livers were harvested from rats at 0, 3, 7, and 28 d of spontaneous recovery from liver fibrosis induced by 4 wk of twice weekly intraperitoneal injections with CCl4. Hydroxyproline analysis and histology of liver sections indicated that the advanced septal fibrosis observed at time 0 (peak fibrosis) was remodeled over 28 d of recovery to levels close to control (untreated liver). alpha-Smooth muscle actin staining of liver sections demonstrated a 12-fold reduction in the number of activated HSC over the same time period with evidence of HSC apoptosis. Ribonuclease protection analysis of liver RNA extracted at each recovery time point demonstrated a rapid decrease in expression of the collagenase inhibitors TIMP-1 and TIMP-2, whereas collagenase mRNA expression remained at levels comparable to peak fibrosis. Collagenase activity in liver homogenates increased through recovery. We suggest that apoptosis of activated HSC may vitally contribute to resolution of fibrosis by acting as a mechanism for removing the cell population responsible for both producing fibrotic neomatrix and protecting this matrix from degradation via their production of TIMPs.

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Selected References

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  1. Abdel-Aziz G., Lebeau G., Rescan P. Y., Clément B., Rissel M., Deugnier Y., Campion J. P., Guillouzo A. Reversibility of hepatic fibrosis in experimentally induced cholestasis in rat. Am J Pathol. 1990 Dec;137(6):1333–1342. [PMC free article] [PubMed] [Google Scholar]
  2. Arthur M. J., Friedman S. L., Roll F. J., Bissell D. M. Lipocytes from normal rat liver release a neutral metalloproteinase that degrades basement membrane (type IV) collagen. J Clin Invest. 1989 Oct;84(4):1076–1085. doi: 10.1172/JCI114270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baker A. J., Mooney A., Hughes J., Lombardi D., Johnson R. J., Savill J. Mesangial cell apoptosis: the major mechanism for resolution of glomerular hypercellularity in experimental mesangial proliferative nephritis. J Clin Invest. 1994 Nov;94(5):2105–2116. doi: 10.1172/JCI117565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Benyon R. C., Iredale J. P., Goddard S., Winwood P. J., Arthur M. J. Expression of tissue inhibitor of metalloproteinases 1 and 2 is increased in fibrotic human liver. Gastroenterology. 1996 Mar;110(3):821–831. doi: 10.1053/gast.1996.v110.pm8608892. [DOI] [PubMed] [Google Scholar]
  5. Burt A. D. C. L. Oakley Lecture (1993). Cellular and molecular aspects of hepatic fibrosis. J Pathol. 1993 Jun;170(2):105–114. doi: 10.1002/path.1711700203. [DOI] [PubMed] [Google Scholar]
  6. Carter E. A., McCarron M. J., Alpert E., Isselbacher K. J. Lysyl oxidase and collagenase in experimental acute and chronic liver injury. Gastroenterology. 1982 Mar;82(3):526–534. [PubMed] [Google Scholar]
  7. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  8. De Bleser P. J., Niki T., Xu G., Rogiers V., Geerts A. Localization and cellular sources of activins in normal and fibrotic rat liver. Hepatology. 1997 Oct;26(4):905–912. doi: 10.1002/hep.510260416. [DOI] [PubMed] [Google Scholar]
  9. Evan G. I., Brown L., Whyte M., Harrington E. Apoptosis and the cell cycle. Curr Opin Cell Biol. 1995 Dec;7(6):825–834. doi: 10.1016/0955-0674(95)80066-2. [DOI] [PubMed] [Google Scholar]
  10. Evan G. I., Wyllie A. H., Gilbert C. S., Littlewood T. D., Land H., Brooks M., Waters C. M., Penn L. Z., Hancock D. C. Induction of apoptosis in fibroblasts by c-myc protein. Cell. 1992 Apr 3;69(1):119–128. doi: 10.1016/0092-8674(92)90123-t. [DOI] [PubMed] [Google Scholar]
  11. Fanidi A., Harrington E. A., Evan G. I. Cooperative interaction between c-myc and bcl-2 proto-oncogenes. Nature. 1992 Oct 8;359(6395):554–556. doi: 10.1038/359554a0. [DOI] [PubMed] [Google Scholar]
  12. Fraser A., Evan G. A license to kill. Cell. 1996 Jun 14;85(6):781–784. doi: 10.1016/s0092-8674(00)81005-3. [DOI] [PubMed] [Google Scholar]
  13. Freije J. M., Díez-Itza I., Balbín M., Sánchez L. M., Blasco R., Tolivia J., López-Otín C. Molecular cloning and expression of collagenase-3, a novel human matrix metalloproteinase produced by breast carcinomas. J Biol Chem. 1994 Jun 17;269(24):16766–16773. [PubMed] [Google Scholar]
  14. Friedman S. L. Seminars in medicine of the Beth Israel Hospital, Boston. The cellular basis of hepatic fibrosis. Mechanisms and treatment strategies. N Engl J Med. 1993 Jun 24;328(25):1828–1835. doi: 10.1056/NEJM199306243282508. [DOI] [PubMed] [Google Scholar]
  15. Gavrieli Y., Sherman Y., Ben-Sasson S. A. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol. 1992 Nov;119(3):493–501. doi: 10.1083/jcb.119.3.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Herbst H., Heinrichs O., Schuppan D., Milani S., Stein H. Temporal and spatial patterns of transin/stromelysin RNA expression following toxic injury in rat liver. Virchows Arch B Cell Pathol Incl Mol Pathol. 1991;60(5):295–300. doi: 10.1007/BF02899560. [DOI] [PubMed] [Google Scholar]
  17. Iredale J. P., Benyon R. C., Arthur M. J., Ferris W. F., Alcolado R., Winwood P. J., Clark N., Murphy G. Tissue inhibitor of metalloproteinase-1 messenger RNA expression is enhanced relative to interstitial collagenase messenger RNA in experimental liver injury and fibrosis. Hepatology. 1996 Jul;24(1):176–184. doi: 10.1002/hep.510240129. [DOI] [PubMed] [Google Scholar]
  18. Iredale J. P., Goddard S., Murphy G., Benyon R. C., Arthur M. J. Tissue inhibitor of metalloproteinase-I and interstitial collagenase expression in autoimmune chronic active hepatitis and activated human hepatic lipocytes. Clin Sci (Lond) 1995 Jul;89(1):75–81. doi: 10.1042/cs0890075. [DOI] [PubMed] [Google Scholar]
  19. Iredale J. P., Murphy G., Hembry R. M., Friedman S. L., Arthur M. J. Human hepatic lipocytes synthesize tissue inhibitor of metalloproteinases-1. Implications for regulation of matrix degradation in liver. J Clin Invest. 1992 Jul;90(1):282–287. doi: 10.1172/JCI115850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Li J., Kim C. I., Leo M. A., Mak K. M., Rojkind M., Lieber C. S. Polyunsaturated lecithin prevents acetaldehyde-mediated hepatic collagen accumulation by stimulating collagenase activity in cultured lipocytes. Hepatology. 1992 Mar;15(3):373–381. doi: 10.1002/hep.1840150303. [DOI] [PubMed] [Google Scholar]
  21. Maruyama K., Feinman L., Fainsilber Z., Nakano M., Okazaki I., Lieber C. S. Mammalian collagenase increases in early alcoholic liver disease and decreases with cirrhosis. Life Sci. 1982 Apr 19;30(16):1379–1384. doi: 10.1016/0024-3205(82)90023-6. [DOI] [PubMed] [Google Scholar]
  22. Montfort I., Pérez-Tamayo R., Alvizouri A. M., Tello E. Collagenase of hepatocytes and sinusoidal liver cells in the reversibility of experimental cirrhosis of the liver. Virchows Arch B Cell Pathol Incl Mol Pathol. 1990;59(5):281–289. doi: 10.1007/BF02899415. [DOI] [PubMed] [Google Scholar]
  23. Murawaki Y., Kawasaki H., Burkhardt H. Serum collagenase activity in chronic liver diseases. Pathol Res Pract. 1994 Oct;190(9-10):929–933. doi: 10.1016/S0344-0338(11)80998-2. [DOI] [PubMed] [Google Scholar]
  24. Murawaki Y., Yamada S., Koda M., Hirayama C. Collagenase and collagenolytic cathepsin in normal and fibrotic rat liver. J Biochem. 1990 Aug;108(2):241–244. doi: 10.1093/oxfordjournals.jbchem.a123187. [DOI] [PubMed] [Google Scholar]
  25. Murawaki Y., Yamamoto H., Kawasaki H., Shima H. Serum tissue inhibitor of metalloproteinases in patients with chronic liver disease and with hepatocellular carcinoma. Clin Chim Acta. 1993 Sep 17;218(1):47–58. doi: 10.1016/0009-8981(93)90221-o. [DOI] [PubMed] [Google Scholar]
  26. Muzzillo D. A., Imoto M., Fukuda Y., Koyama Y., Saga S., Nagai Y., Hayakawa T. Clinical evaluation of serum tissue inhibitor of metalloproteinases-1 levels in patients with liver diseases. J Gastroenterol Hepatol. 1993 Sep-Oct;8(5):437–441. doi: 10.1111/j.1440-1746.1993.tb01544.x. [DOI] [PubMed] [Google Scholar]
  27. Okazaki I., Maruyama K. Collagenase activity in experimental hepatic fibrosis. Nature. 1974 Nov 1;252(5478):49–50. doi: 10.1038/252049a0. [DOI] [PubMed] [Google Scholar]
  28. Patel T., Gores G. J. Apoptosis and hepatobiliary disease. Hepatology. 1995 Jun;21(6):1725–1741. doi: 10.1002/hep.1840210635. [DOI] [PubMed] [Google Scholar]
  29. Perez-Tamayo R., Montfort I., Gonzalez E. Collagenolytic activity in experimental cirrhosis of the liver. Exp Mol Pathol. 1987 Dec;47(3):300–308. doi: 10.1016/0014-4800(87)90015-3. [DOI] [PubMed] [Google Scholar]
  30. Rockey D. C., Boyles J. K., Gabbiani G., Friedman S. L. Rat hepatic lipocytes express smooth muscle actin upon activation in vivo and in culture. J Submicrosc Cytol Pathol. 1992 Apr;24(2):193–203. [PubMed] [Google Scholar]
  31. Rojkind M., Dunn M. A. Hepatic fibrosis. Gastroenterology. 1979 Apr;76(4):849–863. [PubMed] [Google Scholar]
  32. Saile B., Knittel T., Matthes N., Schott P., Ramadori G. CD95/CD95L-mediated apoptosis of the hepatic stellate cell. A mechanism terminating uncontrolled hepatic stellate cell proliferation during hepatic tissue repair. Am J Pathol. 1997 Nov;151(5):1265–1272. [PMC free article] [PubMed] [Google Scholar]
  33. Vaux D. L., Strasser A. The molecular biology of apoptosis. Proc Natl Acad Sci U S A. 1996 Mar 19;93(6):2239–2244. doi: 10.1073/pnas.93.6.2239. [DOI] [PMC free article] [PubMed] [Google Scholar]

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