Assay method for antihepatotoxic activity using galactosamine-induced cytotoxicity in primary-cultured hepatocytes

Conditions were investigated to devise an in vitro assay method for antihepatotoxic activity using galactosamine-produced injury in primary-cultured mouse and rat liver cells. Employing 1.5-h preincubated hepatocytes prepared from rats, which were much more sensitive to the hepatotoxin, a satisfactory assay procedure was achieved. Some natural products known to exert liver-protective effects in vivo were subjected to screening by this in vitro assay method to reveal that cynarin, desoxypodophyllotoxin, glycyrrhetinic acid, glycyrrhizin, picroside II, and silybin possessed significant antihepatotoxic activity. The described assay method may be useful for primary screening of antihepatotoxic activity of materials of plant origin. The assay method has a number of advantages including the ability to dispose numerous samples at one time at a low cost, the requirement of small sample sizes, little variation, and good reproducibility of results.