ANTIOXIDANTS CAN DELAY LIVER-CELL MATURATION WHICH IN TURN AFFECTS GAMMA-GLUTAMYL-TRANSPEPTIDASE EXPRESSION

In normal rats just before weaning the majority of hepatocytes are mononucleated diploids, but within days the number of binucleated cells reaches a peak (approximately 50%) before declining again and there is a steady shift of diploid to tetraploid nuclei. When weanling rats were exposed to ethoxyquin (EQ), the conversion of 2N nuclei to 4N and 8N nuclei as measured by flow cytometry was slowed down. The rapid rise in the number of binucleate cells was also delayed, although the long-term effect was an increased number compared with age-matched controls. It appeared that when EQ was present in the diet, significant numbers of diploid hepatocytes undergoing DNA synthesis also underwent mitosis and cytokinesis giving rise to new diploid hepatocytes. However, many hepatocytes from animals maintained on a control diet did not undergo cytokinesis. Thus the slower 'conversion' of 2N to 4N nuclei in treated hepatocytes was due in part to promotion of cytokinesis in diploid cells undergoing DNA synthesis. The ploidy of a cell would be expected to affect gene expression. EQ is a very potent inducer of gamma-glutamyltranspeptidase (GGT), but expression depended on the age of the animals, the length of treatment time and apparently the ploidy status of the liver. In weanling rats treated with EQ for 7 days, >80% of the hepatocytes expressed GGT, while in 42 day old rats similarly treated < 50% were positive for this enzyme. GGT expression was closely correlated with the percentage of 2N nuclei present in hepatocytes, suggesting that it was more easily induced in cells containing these nuclei than in those containing nuclei of higher ploidy. Although butylated hydroxytoluene (BHT), at the same concentration in the diet, had a similar negative effect on weight gain as did EQ, it had no effect on ploidy, nor did it induce GGT to the same extent as EQ.