The goal of this proposal is to investigate hormonal influences which may regulate the activity of liver alcohol dehydrogenase and ethanol metabolism. Studies in vivo have already demonstrated that the activity of liver alcohol dehydrogenase and rates of ethanol elimination are affected by a variety of hormones. The majority of the studies proposed will be done with primary rat hepatocyte culture. This permits direct assessment of the effects of physiological concentrations of hormones, which is very difficult in vivo because of complex hormonal interactions. The effects of testosterone, dihydrotestosterone, triiodothyronine, growth hormone, corticosterone and estradiol will be determined on liver alcohol dehydrogenase activity in hepatocytes harvested from normal rats and corresponding hormone-deficient hepatocytes harvested from castrated, hypophysectomized, adrenalectomized and ovariectomized rats. Interactions between the hormones on the activity of the enzyme will be explored. Corresponding changes in enzyme-protein will be identified by an enzyme-linked immunosorbent assay. In cases of increased enzyme-protein it will be determined if the changes are due to increased synthesis, decreased degradation or a combination of both processes. Rates of ethanol metabolism will be determined whenever changes in the enzyme are obtained. Conversely, the effects of epinephrine and glucagon, which have been shown to increase the rate of ethanol metabolism in hepatocytes, will be determined on alcohol dehydrogenase activity. The influence of chronic exposure of the hepatocytes to ethanol on the documented effects of the hormones on alcohol dehydrogenase and ethanol metabolism will be assessed. The separate and combined effects of ethanol and the hormones on adrenergic receptors as they relate to ethanol metabolism will be explored. A secondary objective is to determine whether or not hypogonadism and testosterone administration, which modify alcohol dehydrogenase activity and ethanol metabolism in rats also modify ethanol elimination in man.
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