Contributions of dietary carbohydrate and ethanol to alterations in liver glycogen levels and glycolytic activity.

Hepatic glycogen levels are decreased in rats as a consequence of chronic ethanol consumption. In earlier studies ethanol (36% of total calories consumed) replaced carbohydrate in the ethanol-containing diet, thus leading to the possibility that the decreases in liver glycogen were a result of limited dietary carbohydrate. In the present study, rats were administered ethanol in low-carbohydrate (LC) or high-carbohydrate (HC) diets to determine if lowered dietary carbohydrate contributes to the decrease in glycogen levels associated with ethanol consumption. The glycogen content of isolated hepatocytes was not different between rats fed LC or HC in control or ethanol-containing diets. Lactate and pyruvate were measured to determine the effects of dietary carbohydrate and ethanol on glycolytic activity, and were not significantly altered by changes in the levels of dietary carbohydrate. However, ethanol-containing diets resulted in decreased concentrations of hepatic glycogen, lactate, and pyruvate as compared with controls in both LC and HC diets. These observations demonstrate that decreases in glycogen content and lactate + pyruvate concentrations are due to chronic ethanol consumption rather than a carbohydrate deficiency, when carbohydrate is maintained above 10% of total calories.

Signal transduction via cannabinoid receptors.

The endocannabinoids anandamide and 2-arachidonoylglycerol are lipid mediators that signal via CB(1) and CB(2) cannabinoid receptors and Gi/o-proteins to inhibit adenylyl cyclase and stimulate mitogen-activated protein kinase. In the brain, CB(1) receptors interact with opioid receptors in close proximity, and these receptors may share G-proteins and effector systems. In the striatum, CB(1) receptors function in coordination with D(1) and D(2) dopamine receptors, and combined stimulation of CB(1)-D(2) receptor heteromeric complexes promotes a unique interaction to stimulate cAMP production. CB(1) receptors also trigger growth factor receptor signaling cascades in cells by engaging in cross-talk or interreceptor signal transmission with the receptor tyrosine kinase (RTK) family. Mechanisms for CB(1) receptor-RTK transactivation can include stimulation of signal transduction pathways regulated by second messengers such as phospholipase C, metalloprotease cleavage of membrane-bound precursor proteins such as epidermal growth factor which activate RTKs, RTK autophosphorylation, and recruitment of non-receptor tyrosine kinases. CB(1) and CB(2) receptors are expressed in peripheral tissues including liver and adipose tissue, and are induced in pathological conditions. Novel signal transduction resulting from endocannabinoid regulation of AMP-regulated kinase and peroxisome proliferator-activated receptors have been discovered from studies of hepatocytes and adipocytes. It can be predicted that drug discovery of the future will be based upon these novel signal transduction mechanisms for endocannabinoid mediators.

Chronic ethanol consumption and liver glycogen synthesis.

Chronic ethanol consumption results in a dramatic decrease in liver glycogen concentrations, which could be related to either a depressed rate of synthesis or an increased rate of breakdown. Earlier studies suggested that there is not an increase in the rate of glycogenolysis as glycogen phosphorylase activities are not elevated. In the present study it was observed that the incorporation of radiolabeled glucose into glycogen was significantly depressed in hepatocytes from ethanol-fed rats under both anaerobic and aerobic conditions. Chronic ethanol consumption decreased the total glycogen synthase (a + b) activity, which correlated closely with a loss in glycogen synthase protein. However, glycogen synthase messenger RNA levels were not depressed, which indicated posttranscriptional modifications affecting both activity and protein levels. The concentration of glucose transporter 1 was also decreased due to ethanol consumption, but glucose transporter 2 levels were not altered. This latter result suggests that glucose transport in the perivenous region of the liver lobule may be decreased in chronic ethanol consumers. The alterations in glucose transport protein and glycogen synthesis observed in this study may contribute to lowered glycogen synthesis, but do not appear to account for the magnitude of the decreases in glycogen levels and rate of synthesis. Indeed, ethanol effects on glycogen metabolism are likely to be exerted at several levels, including posttranslational modulation of enzyme activities.

Effect of chronic ethanol consumption on respiratory and glycolytic activities of rat periportal and perivenous hepatocytes.

Previous studies (Ivester et al., Arch. Biochem. Biophys. 322, 14-21, 1995) have established that periportal and perivenous hepatocytes isolated from ethanol-fed rats demonstrate lower ATP concentrations than those in control preparations when the cells are maintained at very low oxygen tension. In the present investigation, experiments were implemented with periportal and perivenous hepatocytes to determine the effects of chronic ethanol consumption on cellular respiratory and glycolytic activities, since both contribute to maintenance of the energy state of the liver cell. Both periportal and perivenous hepatocytes from ethanol-fed rats demonstrated significantly increased, rather than decreased, respiratory activity when monitored with oxygen concentrations ranging from 16 to 140 microM. Whole liver hepatocytes from control and ethanol-fed animals demonstrated equivalent oxygen utilization, however. Glycolytic activity, monitored by lactate + pyruvate concentrations obtained after both anaerobic and aerobic incubation protocols, was decreased in both cell types from ethanol-fed animals. The glycogen concentrations in freshly isolated periportal and perivenous hepatocytes were also decreased eight- and sevenfold, respectively, as compared with control preparations. Incubation under anaerobic conditions resulted in almost complete depletion of glycogen in both cell types. These observations suggest the possibility that the decreased energy state observed in hepatocytes from ethanol-fed animals is related to a depression in anaerobic glycolysis due to depletion of the endogenous substrate, glycogen.