A possible mechanism for the anti-ketogenic action of alanine in the rat

1. The anti-ketogenic effect of alanine has been studied in normal starved and diabetic rats by infusing l-alanine for 90min in the presence of somatostatin (10μg/kg body wt. per h) to suppress endogenous insulin and glucagon secretion. 2. Infusion of alanine at 3mmol/kg body wt. per h caused a 70±11% decrease in [3-hydroxybutyrate] and a 58±9% decrease in [acetoacetate] in 48h-starved rats. [Glucose] and [lactate] increased, but [non-esterified fatty acid], [glycerol] and [3-hydroxybutyrate]/[acetoacetate] were unchanged. 3. Infusion of alanine at 1mmol/kg body wt. per h caused similar decreases in [ketone body] (3-hydroxybutyrate plus acetoacetate) in 24h-starved normal and diabetic rats, but no change in other blood metabolites. 4. Alanine [3mmol/kg body wt. per h] caused a 72±9% decrease in the rate of production of ketone bodies and a 57±8% decrease in disappearance rate as assessed by [3-14C]acetoacetate infusion. Metabolic clearance was unchanged, indicating that the primary effect of alanine was inhibition of hepatic ketogenesis. 5. Aspartate infusion at 6mmol/kg body wt. per h had similar effects on blood ketone-body concentrations in 48h-starved rats. 6. Alanine (3mmol/kg body wt. per h) caused marked increases in hepatic glutamate, aspartate, malate, lactate and citrate, phosphoenolpyruvate, 2-phosphoglycerate and glucose concentrations and highly significant decreases in [3-hydroxybutyrate] and [acetoacetate]. Calculated [oxaloacetate] was increased 75%. 7. Similar changes in hepatic [malate], [aspartate] and [ketone bodies] were found after infusion of 6mmol of aspartate/kg body wt. per h. 8. It is suggested that the anti-ketogenic effect of alanine is secondary to an increase in hepatic oxaloacetate and hence citrate formation with decreased availability of acetyl-CoA for ketogenesis. The reciprocal negative-feedback cycle of alanine and ketone bodies forms an important non-hormonal regulatory system.

Metabolism of ketone bodies by ovine brain in vivo

Ketosis was produced by intravenous infusion (5 mmol/kg per h) of [3-14C]acetoacetate (sp act 0.5 muCi/mmol) into fed and 7-day-fasted sheep. Changes in arterial and sagittal sinus blood samples. During acetoacetate infusion, there was a significant increase in ketone body uptake (P less than 0.001) and conversion to 14CO2 in both fed and fasted sheep. Changes in arterial concentrations and cerebral removal of various metabolites were investigated by simultaneous collection of arterial and sagittal sinus blood samples. During acetoacetate infusion, there was a significant increase in ketone body uptake (P less than 0.001) and conversion to 14CO2 in both fed and fasted sheep when compared to control periods (saline infusion). The percent conversion of ketone bodies to 14CO2 was slightly higher in fasted sheep (22%) compared to fed sheep (18%). Blood glucose and free fatty acid concentrations were decreased, but there was a significant increase in blood lactic acid and lactic acid production by the brain. The plasma insulin concentration was increased significantly both in fed and fasted animals. These results indicate that ovine brain can utilize ketone bodies irrespective of nutritional state. In addition, ketone bodies stimulated the production of lactate by ovine brain.

Effect of ketone bodies on cardiac metabolism

The effect of acetoacetate infusion on myocardial metabolism was studied in 13 dogs at varying concentrations of acetoacetate. Acetoacetate was extracted by the myocardium at arterial levels of from 1 to 54 mg/100 ml. At arterial levels of above 60 mg/100 ml, extraction of acetoacetate by the heart was very small. Considerable amounts of the infused acetoacetate were reduced to beta-hydroxybutyrate. Acetoacetate inhibited the utilization of free fatty acids by the heart, resulting in a rise in the respiratory quotient of the heart. An increase in the myocardial extraction of lactate occurred at arterial acetoacetate levels of below 34 mg/100 ml. Between 34 and 80 mg/100 ml of acetoacetate levels, myocardial lactate extraction declined; the ketone became the preferred fuel of the myocardium at arterial acetoacetate levels between 34 and 54 mg/100 ml. Arterial glucose levels fell gradually during the experiment, leading to severe hypoglycemia. The negative myocardial balance of pyruvate significantly increased throughout the experiment. Coronary blood flow, heart rate, left ventricular pressure, myocardial contractility, and EKG were not affected significantly by arterial acetoacetate levels ranging from 1 to 80 mg/100 ml.