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We have studied the effects of polyunsaturated fatty acid and its metabolism on the activity, relative synthesis and mRNA levels for rat hepatic glucose-6-phosphate dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (6PGD). Rats were meal-fed high carbohydrate diets containing either no fat, 5% safflower oil or 5% safflower oil + eicosa-5,8,11,14-tetraynoic acid (TYA). Hepatocytes were isolated and used as a source of RNA, de novo radiolabeled protein and postmitochondrial supernatant for enzyme assay. Dietary safflower oil, as a source of linoleic acid, repressed G6PD activity, synthesis and mRNA levels two- to threefold without significantly changing the amount of carbohydrate consumed. Similar but smaller changes were observed for 6PGD. Dietary fat + TYA (an analogue of arachidonate that inhibits normal metabolism of linoleic acid) prevented the fat-dependent lowering of G6PD and 6PGD activity, synthesis and mRNA levels. Our results suggest that a metabolite of linoleic acid regulates the activity of two lipogenic enzymes, G6PD and 6PGD, by lowering gene expression or mRNA processing or stability.

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Conditions were identified which, for the first time, demonstrate that primary hepatocytes can express the same range of glucose-6-phosphate dehydrogenase (G6PD) synthesis and mRNA as in live rats. Primary hepatocytes were cultured without prior exposure to serum, hormones or carbohydrates. Five modulators implicated in G6PD induction in vivo were examined: insulin, dexamethasone, tri-iodothyronine (T3), glucose and fructose, T3 did not affect G6PD activity, and did not interact with carbohydrate to affect the activity of G6PD. Neither glucose nor fructose alone affected G6PD activity, and they did not interact with insulin to increase G6PD activity. Hepatocytes isolated from fasted rats and cultured in serum-free media with amino acids ad the only energy source how a 12-fold increase in G6PD synthesis and mRNA (measured by a solution-hybridization assay). This induction does not require added hormones or carbohydrate. The addition of insulin alone caused another increase in G6PD synthesis and mRNA. There are at least three distinct phases to G6PD induction under these conditions. The largest increase in G6PD synthesis (12-fold) occurs in the absence of any hormones and with amino acids as the only energy source. This phase is due to increased G6PD mRNA. Insulin causes an additional 2-3-fold increase in G6PD synthesis and mRNA. However, dexamethasone and insulin are both required before G6PD synthesis is equal to that in rats which are fasted and refed on a high-carbohydrate diet.