Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1990 Apr;87(8):3137–3141. doi: 10.1073/pnas.87.8.3137

Interleukin 6 regulates metallothionein gene expression and zinc metabolism in hepatocyte monolayer cultures.

J J Schroeder 1, R J Cousins 1
PMCID: PMC53849  PMID: 2326272

Abstract

Attention has focused on the cytokine interleukin 6 (IL-6) as a major mediator of acute-phase protein synthesis in hepatocytes in response to infection and tissue injury. We have evaluated the effects of IL-6 and IL-1 alpha as well as extracellular zinc and glucocorticoid hormone on metallothionein gene expression and cellular zinc accumulation in rat hepatocyte monolayer cultures. Further, we have evaluated the teleological basis for cytokine mediation by examining cytoprotection from CCl4-induced damage. Incubation of hepatocytes with IL-6 led to concentration-dependent and time-dependent increases in metallothionein-1 and -2 mRNA and metallothionein protein. The level of each was increased within 3 hr after the addition of IL-6 at 10 ng/ml (10 hepatocyte-stimulating factor units/ml). Maximal increases in metallothionein mRNA and metallothionein protein were achieved after 12 hr and 36 hr, respectively. In contrast, IL-1 alpha concentrations as high as 20 ng/ml (1000 lymphocyte-activating factor units/ml) had no effect. Concomitant with the up-regulation of metallothionein gene expression, IL-6 also increased cellular zinc. Responses to IL-6 required the synthetic glucocorticoid hormone dexamethasone and were optimized by increased extracellular zinc. In addition, IL-6 with dexamethasone, dexamethasone alone, and increased extracellular zinc each reversed, in decreasing potency, the deleterious effects of CCl4 on hepatocyte viability as measured by cell protein and lactate dehydrogenase activity of the medium. Thus, IL-6 is a major cytokine mediator of metallothionein gene expression and zinc metabolism in hepatocytes and provides cytoprotection from CCl4-induced hepatotoxicity via a mode consistent with dependence upon increased cellular metallothionein synthesis and zinc accumulation.

Full text

PDF
3137

Images in this article

3139Image
on p.3139

Selected References

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

  1. AMADOR E., DORFMAN L. E., WACKER W. E. SERUM LACTIC DEHYDROGENASE ACTIVITY: AN ANALYTICAL ASSESSMENT OF CURRENT ASSAYS. Clin Chem. 1963 Aug;12:391–399. [PubMed] [Google Scholar]
  2. Abel J., de Ruiter N. Inhibition of hydroxyl-radical-generated DNA degradation by metallothionein. Toxicol Lett. 1989 May;47(2):191–196. doi: 10.1016/0378-4274(89)90075-1. [DOI] [PubMed] [Google Scholar]
  3. Amrani D. L., Mauzy-Melitz D., Mosesson M. W. Effect of hepatocyte-stimulating factor and glucocorticoids on plasma fibronectin levels. Biochem J. 1986 Sep 1;238(2):365–371. doi: 10.1042/bj2380365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Andersen R. D., Taplitz S. J., Birren B. W., Bristol G., Herschman H. R. Rat metallothionein multigene family. Experientia Suppl. 1987;52:373–384. doi: 10.1007/978-3-0348-6784-9_35. [DOI] [PubMed] [Google Scholar]
  5. Andus T., Geiger T., Hirano T., Kishimoto T., Tran-Thi T. A., Decker K., Heinrich P. C. Regulation of synthesis and secretion of major rat acute-phase proteins by recombinant human interleukin-6 (BSF-2/IL-6) in hepatocyte primary cultures. Eur J Biochem. 1988 Apr 15;173(2):287–293. doi: 10.1111/j.1432-1033.1988.tb13997.x. [DOI] [PubMed] [Google Scholar]
  6. Bauer J., Lengyel G., Bauer T. M., Acs G., Gerok W. Regulation of interleukin-6 receptor expression in human monocytes and hepatocytes. FEBS Lett. 1989 May 22;249(1):27–30. doi: 10.1016/0014-5793(89)80008-0. [DOI] [PubMed] [Google Scholar]
  7. Baumann H., Jahreis G. P., Sauder D. N., Koj A. Human keratinocytes and monocytes release factors which regulate the synthesis of major acute phase plasma proteins in hepatic cells from man, rat, and mouse. J Biol Chem. 1984 Jun 10;259(11):7331–7342. [PubMed] [Google Scholar]
  8. Bettger W. J., O'Dell B. L. A critical physiological role of zinc in the structure and function of biomembranes. Life Sci. 1981 Mar 30;28(13):1425–1438. doi: 10.1016/0024-3205(81)90374-x. [DOI] [PubMed] [Google Scholar]
  9. Blalock T. L., Dunn M. A., Cousins R. J. Metallothionein gene expression in rats: tissue-specific regulation by dietary copper and zinc. J Nutr. 1988 Feb;118(2):222–228. doi: 10.1093/jn/118.2.222. [DOI] [PubMed] [Google Scholar]
  10. Castell J. V., Gómez-Lechón M. J., David M., Hirano T., Kishimoto T., Heinrich P. C. Recombinant human interleukin-6 (IL-6/BSF-2/HSF) regulates the synthesis of acute phase proteins in human hepatocytes. FEBS Lett. 1988 May 23;232(2):347–350. doi: 10.1016/0014-5793(88)80766-x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Chvapil M., Ludwig J. C., Sipes I. G., Misiorowski R. L. Inhibition of NADPH oxidation and related drug oxidation in liver microsomes by zinc. Biochem Pharmacol. 1976 Aug 1;25(15):1787–1791. doi: 10.1016/0006-2952(76)90417-2. [DOI] [PubMed] [Google Scholar]
  13. Coppen D. E., Richardson D. E., Cousins R. J. Zinc suppression of free radicals induced in cultures of rat hepatocytes by iron, t-butyl hydroperoxide, and 3-methylindole. Proc Soc Exp Biol Med. 1988 Oct;189(1):100–109. doi: 10.3181/00379727-189-42786. [DOI] [PubMed] [Google Scholar]
  14. Cousins R. J. Absorption, transport, and hepatic metabolism of copper and zinc: special reference to metallothionein and ceruloplasmin. Physiol Rev. 1985 Apr;65(2):238–309. doi: 10.1152/physrev.1985.65.2.238. [DOI] [PubMed] [Google Scholar]
  15. Cousins R. J., Dunn M. A., Leinart A. S., Yedinak K. C., DiSilvestro R. A. Coordinate regulation of zinc metabolism and metallothionein gene expression in rats. Am J Physiol. 1986 Dec;251(6 Pt 1):E688–E694. doi: 10.1152/ajpendo.1986.251.6.E688. [DOI] [PubMed] [Google Scholar]
  16. Cousins R. J., Leinart A. S. Tissue-specific regulation of zinc metabolism and metallothionein genes by interleukin 1. FASEB J. 1988 Oct;2(13):2884–2890. doi: 10.1096/fasebj.2.13.2458983. [DOI] [PubMed] [Google Scholar]
  17. DiSilvestro R. A., Cousins R. J. Glucocorticoid independent mediation of interleukin-1 induced changes in serum zinc and liver metallothionein levels. Life Sci. 1984 Nov 19;35(21):2113–2118. doi: 10.1016/0024-3205(84)90510-1. [DOI] [PubMed] [Google Scholar]
  18. Dinarello C. A. Biology of interleukin 1. FASEB J. 1988 Feb;2(2):108–115. [PubMed] [Google Scholar]
  19. Dunn M. A., Cousins R. J. Kinetics of zinc metabolism in the rat: effect of dibutyryl cAMP. Am J Physiol. 1989 Mar;256(3 Pt 1):E420–E430. doi: 10.1152/ajpendo.1989.256.3.E420. [DOI] [PubMed] [Google Scholar]
  20. Eaton D. L., Toal B. F. Evaluation of the Cd/hemoglobin affinity assay for the rapid determination of metallothionein in biological tissues. Toxicol Appl Pharmacol. 1982 Oct;66(1):134–142. doi: 10.1016/0041-008x(82)90068-0. [DOI] [PubMed] [Google Scholar]
  21. Etzel K. R., Shapiro S. G., Cousins R. J. Regulation of liver metallothionein and plasma zinc by the glucocorticoid dexamethasone. Biochem Biophys Res Commun. 1979 Aug 28;89(4):1120–1126. doi: 10.1016/0006-291x(79)92124-7. [DOI] [PubMed] [Google Scholar]
  22. Failla M. L., Cousins R. J. Zinc accumulation and metabolism in primary cultures of adult rat liver cells. Regulation by glucocorticoids. Biochim Biophys Acta. 1978 Oct 18;543(3):293–304. doi: 10.1016/0304-4165(78)90047-8. [DOI] [PubMed] [Google Scholar]
  23. Failla M. L., Cousins R. J. Zinc uptake by isolated rat liver parenchymal cells. Biochim Biophys Acta. 1978 Feb 1;538(3):435–444. doi: 10.1016/0304-4165(78)90405-1. [DOI] [PubMed] [Google Scholar]
  24. Girotti A. W., Thomas J. P., Jordan J. E. Inhibitory effect of zinc(II) on free radical lipid peroxidation in erythrocyte membranes. J Free Radic Biol Med. 1985;1(5-6):395–401. doi: 10.1016/0748-5514(85)90152-7. [DOI] [PubMed] [Google Scholar]
  25. Hamer D. H. Metallothionein. Annu Rev Biochem. 1986;55:913–951. doi: 10.1146/annurev.bi.55.070186.004405. [DOI] [PubMed] [Google Scholar]
  26. Huber K. L., Cousins R. J. Maternal zinc deprivation and interleukin-1 influence metallothionein gene expression and zinc metabolism of rats. J Nutr. 1988 Dec;118(12):1570–1576. doi: 10.1093/jn/118.12.1570. [DOI] [PubMed] [Google Scholar]
  27. Ikebuchi K., Wong G. G., Clark S. C., Ihle J. N., Hirai Y., Ogawa M. Interleukin 6 enhancement of interleukin 3-dependent proliferation of multipotential hemopoietic progenitors. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9035–9039. doi: 10.1073/pnas.84.24.9035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Imbra R. J., Karin M. Metallothionein gene expression is regulated by serum factors and activators of protein kinase C. Mol Cell Biol. 1987 Apr;7(4):1358–1363. doi: 10.1128/mcb.7.4.1358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Karin M., Imbra R. J., Heguy A., Wong G. Interleukin 1 regulates human metallothionein gene expression. Mol Cell Biol. 1985 Oct;5(10):2866–2869. doi: 10.1128/mcb.5.10.2866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Koj A., Gauldie J., Regoeczi E., Sauder D. N., Sweeney G. D. The acute-phase response of cultured rat hepatocytes. System characterization and the effect of human cytokines. Biochem J. 1984 Dec 1;224(2):505–514. doi: 10.1042/bj2240505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  32. Ludwig J. C., Misiorowski R. L., Chvapil M., Seymour M. D. Interaction of zinc ions with electron carrying coenzymes NADPH and NADH. Chem Biol Interact. 1980 Apr;30(1):25–34. doi: 10.1016/0009-2797(80)90111-8. [DOI] [PubMed] [Google Scholar]
  33. Morrone G., Ciliberto G., Oliviero S., Arcone R., Dente L., Content J., Cortese R. Recombinant interleukin 6 regulates the transcriptional activation of a set of human acute phase genes. J Biol Chem. 1988 Sep 5;263(25):12554–12558. [PubMed] [Google Scholar]
  34. Nebes V. L., DeFranco D., Morris S. M., Jr Cyclic AMP induces metallothionein gene expression in rat hepatocytes but not in rat kidney. Biochem J. 1988 Oct 15;255(2):741–743. [PMC free article] [PubMed] [Google Scholar]
  35. Thomas J. P., Bachowski G. J., Girotti A. W. Inhibition of cell membrane lipid peroxidation by cadmium- and zinc-metallothioneins. Biochim Biophys Acta. 1986 Dec 10;884(3):448–461. doi: 10.1016/0304-4165(86)90195-9. [DOI] [PubMed] [Google Scholar]
  36. Thornalley P. J., Vasák M. Possible role for metallothionein in protection against radiation-induced oxidative stress. Kinetics and mechanism of its reaction with superoxide and hydroxyl radicals. Biochim Biophys Acta. 1985 Jan 21;827(1):36–44. doi: 10.1016/0167-4838(85)90098-6. [DOI] [PubMed] [Google Scholar]
  37. Woloski B. M., Smith E. M., Meyer W. J., 3rd, Fuller G. M., Blalock J. E. Corticotropin-releasing activity of monokines. Science. 1985 Nov 29;230(4729):1035–1037. doi: 10.1126/science.2997929. [DOI] [PubMed] [Google Scholar]
  38. Zhang Y. H., Lin J. X., Yip Y. K., Vilcek J. Enhancement of cAMP levels and of protein kinase activity by tumor necrosis factor and interleukin 1 in human fibroblasts: role in the induction of interleukin 6. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6802–6805. doi: 10.1073/pnas.85.18.6802. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES