ESTROGEN-RECEPTOR LEVELS AND OCCUPANCY IN HEPATIC SINUSOIDAL ENDOTHELIAL AND KUPFFER CELLS ARE ENHANCED BY INITIATION WITH DIETHYLNITROSAMINE AND PROMOTION WITH 17-ALPHA-ETHINYLESTRADIOL IN RATS

We report the presence of estrogen receptors (ER) in rat liver sinusoi dal endothelial (SEC) and Kupffer cells (KC), which exhibited comparab le saturation kinetics and receptor affinity (K-d) for 17 alpha-estrad iol, as characterized for the rat hepatocyte ER, The ER levels in both cell types were significantly decreased by ovariectomy, indicating a regulatory role of estrogens, Initiation of ovariectomized rats with a single dose (200 mg/kg) of diethylnitrosamine (DEN) or saline (S), fo llowed by chronic exposure to 17 alpha-ethinylestradiol (EE(2)), 90 mu g/kg/day for 30 weeks, packed in cholesterol (C) resulted in signific ant changes of ER levels in both the endothelial and Kupffer cells. Th e isolation of enriched liver SEC and KC populations by centrifugal el utriation allowed for the evaluation of chronic EE(2) exposure and DEN -induced alterations on each cell type. The DEN-EE(2) regime significa ntly enhanced gamma-glutamyltranspeptidase activity in SEC (5-fold) an d KC (6.6-fold) compared to the S/C treated animals. Nuclear ER levels were elevated 5.1-fold in the SEC and 6.5-fold in the KC, and both ce ll types exhibited significant increases in the proportion of occupied nuclear ER compared to the S/C derived cells, suggesting that exogeno us estrogens could influence SEC and KC function through changes in ER levels and occupancy, ER occupancy was similar to 50 % of the total E R in SEC and KC from DEN-EE(2) rats. Increases in ER and occupancy for SEC and KC were similar to those observed for hepatocytes, Cellular g rowth was clearly modified in DEN-EE(2) animals as indicated by a 4- t o 10-fold increase in the proportion of SEC, KC or hepatocytes in S-ph ase as shown by flow cytometry, However, unlike hepatocytes, the epide rmal growth factor receptor (EGFR) was not detected in SEC or KC using a monoclonal EGFR antibody. These findings suggest that the EGFR at 3 0 weeks is not involved in EE(2)-mediated stimulation of mitogenesis i n SEC and KC which may be different from hepatocytes. In summary, our studies demonstrate that SEC and KC contain significant amounts of hig h-affinity ER and that ER pathways may modulate some activities of the SEC and KC, but that ER-EGFR interactions may be different in these c ells from hepatocytes.