Summary
Sodium butyrate (NaB), a 4-carbon fatty acid, has been reported to activate the metallothionein (MT) gene in certain carcinoma cell lines. Because the effects of NaB are dependent on the cell type investigated, this study was conducted to determine if NaB and its homologues induce MT in rat primary hepatocyte cultures. Hepatocytes were grown on monolayer for 12 h and subsequently treated with formate, acetate, propionate (NaP), NaB, and valeric acid for 10 to 58 h. To examine their interaction with known MT inducers, cadmium (Cd), zinc (Zn), or dexamethasone (Dex) were added to some cultures. MT protein in the cells was quantitated by the Cd-hemoglobin assay; MT-1 mRNA was analyzed by Northern blot hybridizations with oligonucleotide probes, and quantitated by slot-blot analysis. Among the 1 to 5 carbon carboxylic acids, only NaP (3 carbon) and NaB (4 carbon) induced MT. NaP and NaB alone produced a moderate increase in MT two- to fourfold over control), but when combined with Cd or Dex, an additive increase was observed. However, when combined with Zn, a synergistic increase was detected. NaB and Zn synergistically increased MT protein, but produced only an additive increase in MT mRNA, suggesting the involvement of some posttranscriptional event(s) in the NaB-Zn induction of MT. In conclusion, NaP and NaB induced MT in normal cultured rat hepatocytes, producing an additive increase in MT protein with Cd and Dex, and a synergistic increase in MT protein with Zn.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Andersen, R. D.; Birren, B. W.; Taplitz, S. J., et al. Rat metallothionein-1 structural gene and three pseudogenes, one of which contain 5’-regulatory sequence. Mol. Cell. Biol. 6:302–314; 1986.
Andrews, G. K.; Adamson, E. D. Butyrate selectively activates the metallothionein gene in teratocarcinoma cells and induces hypersensitivity to metal induction. Nucleic Acids Res. 15:5461–5475; 1987.
Andrews, G. K. Regulation of metallothionein gene expression. Recent Prog. Food Nutr. Sci. 14:193–258; 1990.
Berry, M. N.; Friend, D. S. High-yield preparation of isolated rat liver parenchymal cells. J. Cell Biol. 43:506–520; 1969.
Birren, B. W.; Herschman, H. R. Regulation of the rat metallothionein-I gene by sodium butyrate. Nucleic Acids Res. 14:853–867; 1986.
Birren, B. W.; Taplitz, S. J.; Herschman, H. R. Butyrate-induced changes in nuclease sensitivity of chromatin cannot be correlated with transcriptional activation. Mol. Cell. Biol. 7:3863–3870; 1987.
Bissell, D. M.; Guzelian, P. S. Phenotypic stability of adult rat hepatocytes in primary monolayer culture. Ann. NY Acad. Sci. 349:85–98; 1980.
Bracken, W. M.; Klaassen, C. D. Induction of metallothionein in rat primary hepatocyte cultures: evidence for direct and indirect induction. J. Toxicol. Environ. Health 22:163–174; 1987.
Bradford, M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72:248–254; 1976.
Cherian, M. G., Nordberg, M. Cellular adaption in metal toxicology and metallothionein. Toxicology 28:1–15; 1983.
Chomczynski, P.; Sacchi, N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162:156–159; 1987.
Collins, M. L.; Hunsaker, W. R. Improved hybridization assays employing tailed oligonucleotide probes: a direct comparison with 5’-end-labeled oligonucleotide probes and nick-translated plasmid probes. Anal. Biochem. 151:211–224; 1985.
Cousins, R. J. Absorption, transport, and hepatic metabolism of copper and zinc: special reference to metallothionein and ceruloplasmin. Physiol. Rev. 65:238–309; 1985.
Dunn, M. A.; Blalock, T. L.; Cousins, R. J. Metallothionein. Proc. Soc. Exp. Biol. Med. 185:107–119; 1987.
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. 66:134–142; 1982.
Engelmann, G. L.; Staecker, J. L.; Richardson, A. G. Effect of sodium butyrate on primary cultures of adult rat hepatocytes. In Vitro Cell. Dev. Biol. 23:86–92; 1987.
Guguen-Guillouzo, C.; Guillouzo, A. Modulation of functional activities in cultured rat hepatocytes. Mol. Cell. Biochem. 53/54:35–56; 1983.
Kreamer, B. L.; Staecker, J. L.; Sawada, N., et al. Use of a low-speed, iso-density percoll centrifugation method to increase the viability of isolated rat hepatocyte preparations. In Vitro Cell. Dev. Biol. 22:201–211; 1986.
Kruh, J. Effects of sodium butyrate, a new pharmacological agent, on cells in culture. Mol. Cell. Biochem. 42:65–82; 1982.
Lehrach, H.; Diamond, D.; Wozney, J. M., et al. RNA molecular weight determination by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry 16:4743–4751; 1977.
Liu, J.; Kershaw, W. C.; Klaassen, C. D. Rat primary hepatocyte cultures are a good model for examining metallothionein-induced tolerance to cadmium toxicity. In Vitro Cell. Dev. Biol. 26:75–79; 1990.
Nakamura, T.; Ichihara, A. Control of growth and expression of differentiated functions of mature hepatocytes in primary culture. Cell Struct. Funct. 10:1–16; 1985.
Prasad, K. N.; Sinha, P. K. Effect of sodium butyrate on mammalian cells in culture: a review. In Vitro 12:125–132; 1976.
Prasad, K. N. Butyric acid: a small fatty acid with diverse biological functions. Life Sci. 27:1351–1358; 1980.
Sambrook, J.; Fritsch, E. F.; Maniatis, T., editors. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press; 1989:7.3–7.87.
Staecker, J. L.; Sawada, N.; Pitot, H. C. Stimulation of DNA synthesis in primary cultures of adult rat hepatocytes by sodium butyrate. Biochem. Biophys. Res. Commun. 147:78–85; 1987.
Staecker, J. L.; Sattler, C. A.; Pitot, H. C. Sodium butyrate preserves aspects of the differentiated phenotype of normal adult rat hepatocytes in culture. J. Cell. Physiol. 135:356–376; 1988.
Steele, R. G. D.; Torrie, J. H. Principles and procedures of statistics. New York: McGraw-Hill Book Co.; 1960:99–160.
Thomas, D. J.; Angle, C. R.; Swanson, S. A., et al. Effect of sodium butyrate on metallothionein induction and cadmium cytotoxicity in ROS 17/2.8 ceUs. Toxicology 66:35–46; 1991.
Thome, A. W.; Kmiciek, D.; Mitchelson, K., et al. Patterns of histone acetylation. Eur. J. Biochem. 193:701–713; 1990.
Waalkes, M. P.; Wilson, M. J. Enhancement of cadmium-induced metallothionein synthesis in cultured TRL 1215 cells by butyric acid pretreatment. Toxicol. Lett. 32:289–294; 1986.
Webb, M. Role of metallothionein in cadmium metabolism. In: Foulkes, E. C., ed. Cadmium (Handbook of experimental pharmacology, vol. 80). Berlin, Heidelberg: Springer-Verlag Press; 1986:281–328.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Liu, J., McKIM, J.M., Liu, Y.P. et al. Effects Of Butyrate Homologues On Metallothionein Induction In Rat Primary Hepatocyte Cultures. In Vitro Cell Dev Biol - Animal 28, 320–326 (1992). https://doi.org/10.1007/BF02877055
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02877055