We reported recently that heparin-binding EGF-like growth factor (HB-EGF), a member of the EGF family, is a new hepatotrophic factor for the regeneration of rat liver after partial hepatectomy. The current study examined changes in the amount of HB-EGF mRNA in liver injured by hepatotoxins. The level of HB-EGF mRNA was very low in normal rat liver, but increased markedly in the liver of rats injured by CCl4, showing two peaks, the first at 6 h and the second at 36 h. Western blot analysis showed that HB-EGF protein in the liver of CCl4-treated rats (at 6 h) was increased about 3.4-fold above normal. The level of HB-EGF mRNA also increased markedly in the liver of rats treated with D-galactosamine, showing a major peak at 18 h, and a smaller one at 6 h. These results indicate that HB-EGF may play a role in the regeneration of the liver following hepatotoxic injury.