Effects of butyrate homologues on metallothionein induction in rat primary hepatocyte cultures.

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.