Selected opioids, ethanol and intake of ethanol

Rats were given an opportunity to drink tap water or a sweetened ethanol solution once a day. Across initial days of opportunity, rats increased their intake of the ethanol solution. Prior to some days' sessions with presented fluids, rats received either an injection of placebo (the carrier of drugs) or doses of ethylketocyclozocine, diprenorphine, or ethanol. Diprenorphine increased rats' intake of the ethanol solution compared to placebo. The other agents did not reliably modify intakes. These findings support a conclusion that selected activity in opioid systems of brain increase the propensity to drink alcoholic beverages.     

Tests of opioid deficiency hypotheses of alcoholism.

Eighty-three male rats were maintained on a daily regimen involving 22 h of deprivation of fluids followed by 2 h of access to water and a sweetened alcoholic beverage (12% ethanol, 5% sucrose). After about 3 weeks of such a regimen, rats eventually take considerable amounts of ethanol daily. In the present study, a series of injections of opioids was given subsequent to establishing stable daily intakes of ethanol. Specifically, before a day's opportunity to take fluids, some rats were given a small dose of morphine (2.0 mg/kg), while others were given a dose of naloxone (4.0 mg/kg). When morphine was given 0.5 h before the opportunity to drink, intake of ethanol was increased. However, when morphine was given 4.0 h before the opportunity, intake of ethanol was decreased. Nearly opposite effects were observed when naloxone was given. Other experiments tested the effects of giving morphine 4.0 h before the opportunity to drink plus the effects of a small dose of naloxone or plus the effects of a small dose of morphine. Morphine given 4.0 h before potentiated the effects of a small dose of naloxone and attenuated the effects of another dose of morphine. The effects of morphine were also shown to be similar among rats taking a solution of ethanol and water rather than a sweetened solution. These data provide support for the idea that surfeits, not deficits, in opioidergic activity increase propensity to take alcoholic beverages.     

Constant infusions of morphine and intakes of sweetened ethanol solution among rats

Previous studies have indicated that injections of small doses of morphine increase rats' intake of solutions containing ethanol when rats have a choice of either water or a solution containing ethanol. In this experiment, rats which were implanted with osmotic pumps that delivered constant infusions of morphine (0.6 mg/kg/hr across 24 days) had elevated daily intakes of ethanol, as compared to controls, from the second day of opportunity to take the alcoholic beverage until the pumps were removed. In addition, half of the rats with pumps infusing morphine also received injections of morphine (1.0 mg/kg) just before the 1.5-hr opportunity to take alcoholic beverage or water every day for 8 days. Across this 8-day period, these rats took a mean of 5.18 g of pure ethanol/kg of body weight (g/kg) during the 1.5-hr opportunity to take the alcoholic beverage. This was reliably more than the mean of 4.02 g/kg that their counterparts (having morphine pumps and receiving injections of saline) took across the same period. These data support the hypothesis that a surfeit of opioidergic ligand may potentiate drinking of alcoholic beverages.     

Selected opioids modify intake of sweetened ethanol solution among female rats

Water-deprived female rats were given a daily, 1.5-hr opportunity to take either a sweetened ethanol solution or water. Across days, they increased their intake of ethanol solution and had stable intakes of about 2 g of pure ethanol/kg after 3 weeks. Morphine (1.0 mg/kg) alone, and in combination with diprenorphine (25 micrograms/kg), increased intake of ethanol solution among females similar to the increased intake seen with males under similar procedures. Fentanyl dose-relatedly increased intake of ethanol. The data strengthen the idea that one or more of the endogenous opioid systems, but not all, are involved with instances of "excessive" intake of alcoholic beverages.     

Persistence and specificity of small doses of morphine on intake of alcoholic beverages

Subsequent to water deprivation, male rats were given daily, 1.5-hr opportunities to take either water or a sweetened ethanol solution (ES). Each day, 15 min before the session, rats received a subcutaneous injection of either morphine (1.0 mg/kg) or saline. Across daily sessions, rats given saline gradually increased their intake of ES, until they were eventually taking about 2.0 to 3.0 g of ethanol/kg a session. Rats receiving morphine took greater amounts of ES from nearly the first opportunities. Additional tests assessed the effects of small doses of morphine on intakes of some sucrose solutions, and sweetened solutions containing methanol or propanol. The data support the conclusion that small doses of morphine persistently increase intake of ES across many days (up to 100) of testing, but that the effect is not unique to ES. Even though morphine's effects are not specific to ethanol, the fact that morphine persistently increases intake of ES is of interest with respect to theories of alcoholism.     

Endogenous opioids and alcohol dependence: opioid alkaloids and the propensity to drink alcoholic beverages

Rats consume alcoholic beverages in a wide variety of circumstances. Opioid antagonists, naloxone and naltrexone, decrease intake of many ingesta, including alcoholic beverages. Small doses of morphine increase intake of alcoholic beverages. Further, the effects of small doses of morphine are persistent and there is no sign that tolerance to morphine's ability to increase alcohol intake develops as seen with morphine's ability to produce analgesia. Morphine's effects can combine with other variables that enhance intake of alcoholic beverages to produce very large daily intakes of ethanol. These generalizations, from a large number of separate experiments, support the conclusion that alcoholism is a special case of an ingestive disorder involving opioidergic systems.     

Further studies of opioids and intake of sweetened alcoholic beverage

Rats were placed on a daily regimen of water deprivation followed by a limited opportunity to take either water or a sweetened alcoholic beverage. Across days of opportunity, they took less water and more alcohol until they were taking considerable amounts of alcohol. Naloxone, the classic antagonist at opioid receptors, was given prior to opportunity to drink and it reduced intakes of alcoholic beverage. Small doses of morphine, the classic agonist, increased intakes of alcoholic beverage at doses as low as 0.41 mg/kg. The effects of two other antagonists at opioid receptors (LY117413 and MR2266) were also tested. Both reduced intakes of alcohol. The data showing that more than one or two opioid antagonists reduce intakes of alcoholic beverages, even palatable beverages usually taken in large amounts, and that morphine increases intakes at very low doses, strengthen the idea that endogenous opioid systems are involved in modulating intake of alcohol.     

Opioidergic, serotonergic, and dopaminergic manipulations and rats' intake of a sweetened alcoholic beverage.

Groups of rats were maintained on a daily regimen of 22 h of water deprivation followed by a 2-h opportunity to take either water or a sweetened ethanol solution (ES). In one experiment, it was shown that previous morphine (M) dependence had no effect on initial daily intakes of fluids. After stable ES intakes were achieved, a variety of pharmacological manipulations were assessed for their effects on intake of the ES. Nalmefene, an opioid antagonist, dose-relatedly decreased intakes of ES, and was effective across days of injections. Fluoxetine (FX), a serotonergic reuptake inhibitor, also reduced ES intakes dose relatedly, and across days of injections, but the reduction was not as great as that seen with opioid antagonists. A small dose of M increased ES intakes when given in combination with an ineffective dose of FX, just as it does by itself. However, M had no effect on ES intakes in combination with an effective dose of FX. Pimozide (PIM), a dopaminergic antagonist, dose-relatedly decreased intakes of ES and water, and responding for positively reinforcing intracranial stimulation (ICS). When given in combination, M blunted PIM's reduction of ES intake, but had no effect on PIM's ability to decrease either intake of water or responding for ICS. Amphetamine did not reliably affect rats' intakes of ES across a range of doses. The data, in addition to previous work, lead to the idea that endogenous opioid systems are more salient, with respect to intake of alcoholic beverages, than the other tested neurotransmitter systems. Furthermore, the collective data suggest that a long-lasting opioid antagonist may be an effective pharmacological adjunct to other treatments for alcohol abuse and alcoholism.     

Manipulations of the renin-angiotensin system and intake of a sweetened alcoholic beverage among rats.

Standard laboratory rats were maintained on a daily regimen involving deprivation of fluids for 22 h followed by a 2-h opportunity to drink water and a sweetened alcoholic beverage. Angiotensin II, in doses ranging from 0.1 to 1.25 mg/kg, dose relatedly decreased rats' mean intake of ethanol. All doses increased rats' mean intake of water. Angiotensin II, 0.25 mg/kg, reliably reduced intake of ethanol when it was presented alone during the 1st h of the daily 2-h drinking session, and reliably increased intake of water when it was subsequently presented alone during the 2nd h. Thus the reduction in intake of ethanol seen when the alcoholic beverage is presented concurrently with water is probably not merely due to the increase in intake of water. Lisinopril, an angiotensin converting enzyme inhibitor, in doses of 0.3, 1.0, and 3.0 mg/kg, dose relatedly decreased intake of ethanol, but only after several days of injections. Concurrent intake of water was increased dose relatedly. When injections of lisinopril ceased, intakes of both ethanol and water took several days to return to control levels. Pretreatment with lisinopril, 3.0 mg/kg, for 8 days, had no effect on subsequent intakes of either water or ethanol. Lisinopril, 3.0 mg/kg, had no effect on rats' intake of a sweet solution without ethanol. These results confirm previous work and extend the data base supporting the idea that the renin-angiotensin system plays a role in modulating intake of ethanol.     

Morphine and naloxone modulate intake of ethanol

Rats, deprived of food and water for 18 hr, were given an opportunity to drink water and sweetened ethanol solution for 1 hr prior to being fed and watered for 5 hr, daily. One group received water and sucrose solution without ethanol and other groups received water and sucrose solution with 3, 6, 12 or 24% ethanol. Prior to some days' opportunities to drink, rats were injected with morphine (2.5 mg/kg), naloxone (10 mg/kg), or saline. Morphine increased intake of solutions containing ethanol as compared to intake under placebo. Naloxone reduced intakes of both fluids. Since morphine only increased sucrose solution intake when it contained ethanol, it was concluded that increments in opioid activity increase rats' avidity for ethanol.     

Methadone, pentobarbital, pimozide and ethanol-intake

For 28 days, water-deprived rats were given a daily, 1-hr opportunity to take a sweetened ethanol solution (ES) or water. Across days under this regimen, rats gained weight normally and increased intake of ES until they were taking considerable amounts. Across the next 13 days of the regimen, selected groups were given, before the opportunity to drink, one of five doses of methadone (from 0.5 to 8.0 mg/kg), pentobarbital (from 5 to 30 mg/kg), or their vehicles. Large doses of both agents increased intakes, with methadone (1 and 2 mg/kg) increasing only ES-intake. Subsequently, while the daily regimen continued, rats were given pimozide (0.2, 0.4, or 0.6 mg/kg) at either 1 or 4 hr before the opportunity to drink. Pimozide did not reduce ES-intake. Next, they were given a dose of pentobarbital (5.0 mg/kg) with challenge doses of naloxone (0.3, 1.0, 3.0 mg/kg). Naloxone dose-relatedly antagonized pentobarbital's potential to increase intakes.     

Moderate ethanol ingestion, redox status, and cardiovascular system in the rat

Moderate intake of alcoholic beverages decreases the incidence of cardiovascular pathologies, but it is in dispute if cardioprotective effects are due to ethanol, to polyphenolic compounds present in beverages or to a combination of both. In humans, effects of high, moderate, and low doses of alcoholic beverages are widely studied, but effects of pure alcohol remain unclear. On the other hand, experiments with laboratory animals are centered on high toxicological doses of ethanol but not on low doses. In the present study, we have aimed to mimic in the rat the pattern of alcohol intake in Mediterranean population. Alcohol ingestion is spread along the day and not always related to solid food consumption. We tried to define the beneficial and harmful effects of pure ethanol ingestion without polyphenol’s influence. Experimental rats were given 1% ethanol in their drinking water for 30 days, resulting in a daily ingestion of 0.27 mL of ethanol/rat/d. Ethanol ingestion did not cause deleterious effects on the general status of the animals, but it decreased cholesterol, triglycerides, and catecholamine stores’ rate of utilization in peripheral sympathetic system. Moreover, ethanol lowered pulmonary arterial pressure and did not alter systemic arterial pressure. In the liver, the reduced glutathione/oxidized glutathione ratio was augmented and lipid peroxide, superoxide dismutase, and glutathione peroxidase activities were decreased. However, catalase activity was unaltered. Liver cytochrome P4502E1 distribution and protein level and activity were unchanged by ethanol ingestion. Data indicate a lack of harmful effects and underscore a set of potentially beneficial effects of this dose of ethanol.     

DOES THE BLOCKADE OF OPIOID RECEPTORS INFLUENCE THE DEVELOPMENT OF ETHANOL DEPENDENCE?

We have tested whether the opioid antagonists naloxone (2 mg/kg),naltrexone (2 mg/kg) and diprenorphine (0.2 mg/kg), and theagonist morphine (4–8 mg/kg) given subcutaneously (10min before ethanol for 7 days) modify the ethanol withdrawalsyndrome (audiogenic seizures) following chronic ethanol intoxicationin rats. We found that naloxone, naltrexone and diprenorphinemodified the ethanol withdrawal syndrome. These findings donot rule out the possibility of a biochemical link between theaction of ethanol and opiates at the level of opioid receptors.     

Citalopram as an Inhibitor of Voluntary Ethanol Intake in the Male Rat

Psychological dependence was induced in rats by a 1-year intermittent exposure to intoxicating doses of ethanol, and recorded by the rat’s ability to later take the same dose of ethanol independent of the offered concentration. Citalopram (10 or 40 mg/kg/day) was given for 3 weeks with ethanol available only the first and the last day; 10 mg/kg had no effect. On the first treatment day 40 mg/kg decreased ethanol intake. On the last treatment day 40 mg/kg had no effect. The following week the ethanol intake was higher than before the treatment in the 40 mg/kg group. During the four posttreatment weeks the ethanol intake of the 40 mg/kg group dropped significantly. Citalopram was retested 18 weeks after the first treatment during 1 week, with continuous access to ethanol; 10 mg/kg had no effect and 40 mg/kg decreased ethanol intake at day 1, reaching a minimum in day 3. A tolerance to this effect was seen at the end of the week. Thus, in this model an acute dose of citalopram can decrease ethanol intake, but tolerance to this effect develops when citalopram is given both with and without access to ethanol.     

The margin of exposure to formaldehyde in alcoholic beverages

Formaldehyde has been classified as carcinogenic to humans (WHO IARC group 1). It causes leukaemia and nasopharyngeal cancer, and was described to regularly occur in alcoholic beverages. However, its risk associated with consumption of alcohol has not been systematically studied, so this study will provide the first risk assessment of formaldehyde for consumers of alcoholic beverages.Human dietary intake of formaldehyde via alcoholic beverages in the European Union was estimated based on WHO alcohol consumption data and literature on formaldehyde contents of different beverage groups (beer, wine, spirits, and unrecorded alcohol). The risk assessment was conducted using the margin of exposure (MOE) approach with benchmark doses (BMD) for 10 % effect obtained from dose-response modelling of animal experiments.For tumours in male rats, a BMD of 30 mg kg(-1) body weight per day and a "BMD lower confidence limit" (BMDL) of 23 mg kg(-1) d(-1) were calculated from available long-term animal experiments. The average human exposure to formaldehyde from alcoholic beverages was estimated at 8·10(-5) mg kg(-1) d(-1). Comparing the human exposure with BMDL, the resulting MOE was above 200,000 for average scenarios. Even in the worst-case scenarios, the MOE was never below 10,000, which is considered to be the threshold for public health concerns.The risk assessment shows that the cancer risk from formaldehyde to the alcohol-consuming population is negligible and the priority for risk management (e.g. to reduce the contamination) is very low. The major risk in alcoholic beverages derives from ethanol and acetaldehyde.     

Blockade of gamma-aminobutyric acid receptors does not modify the inhibiton of ethanol intake induced by Hypericum perforatum in rats

AIMS: Recent studies have shown that Hypericum perforatum extracts (HPE) inhibit ethanol intake in alcohol-preferring rats, but their mechanism of action is still unknown. HPE have been shown to bind at gamma-aminobutyric acid (GABA)(A) and GABA(B) receptors, to inhibit GABA reuptake, to evoke GABA release from synaptosomes and to exert an anxiolytic effect that is blocked by the benzodiazepine antagonist flumazenil. Since GABA-ergic mechanisms are known to influence ethanol intake, the present study was aimed at investigating whether they might mediate the effect of a CO2 Hypericum extract (HPCO2) on ethanol intake in genetically selected Marchigian Sardinian alcohol-preferring (msP) rats. METHODS: The GABA(A) receptor antagonist bicuculline and the GABA(B) receptor antagonists CGP-36742 and phaclofen were tested versus the effect of HPCO2 on ethanol intake. RESULTS: The results of the present study confirm that HPCO2, given by intragastric injection, markedly reduces ethanol intake in msP rats and its effect is behaviourally selective, since the same doses which inhibited ethanol intake did not modify the simultaneous intake of food or water. The GABA(A) receptor antagonist bicuculline, given by intraperitoneal (i.p.) injection at a dose of 2 mg/kg, which effectively antagonizes the effects of GABA(A) receptor agonists, did not modify the effect of HPCO2, 15 or 125 mg/kg. The GABA(B) receptor antagonists CGP-36742, given by i.p. injection at a dose of 100 mg/kg, and phaclofen, given by intracerebroventricular injection at a dose of 25 micro g/rat, did not modify the inhibitory effect on alcohol intake induced by HPCO2, 15 or 125 mg/kg. The same doses of the two GABA(B) receptor antagonists induced a pronounced reduction of the effect of the GABA(B) receptor agonist bacoflen, given by i.p. injection at a dose of 5 mg/kg. CONCLUSIONS: These findings suggest that the inhibitory effects of HPE on ethanol intake are not mediated by GABA agonist actions.     

Morphine increases intake of beer among rats

Rats were maintained on a daily regimen of 22 h of water deprivation followed by a 2-h opportunity to take water and sweetened alcoholic beverage containing 12% ethanol. After 30 days, the alcoholic beverage was changed to beer containing either 3% or 6% ethanol. After 20 daily sessions with beer, they received, before the next session, an injection of saline. On the next day, they received a 1.0 mg/kg injection of morphine before the session. Morphine reliably increased rats' mean intake of both kinds of beer. Subsequently, the concentration of ethanol in each groups' beer was changed. The 3% group's beer was switched to 6%, and the 6% group's to 3%. Both groups altered their mean intake of beer in an apparent attempt to maintain intakes of nearly the same amount of ethanol, but presentation of 6% beer resulted in greater intakes of ethanol.     

Nicotine increases ethanol preference but decreases locomotor activity during the initial stages of chronic ethanol withdrawal

AIM: The ability of nicotine to modify withdrawal symptoms in rats chronically treated with alcohol, with respect to locomotor activity and ethanol or nicotine preference, has been evaluated in these studies. METHODS AND RESULTS: Preliminary studies showed that locomotor activity increased 8-9 h after withdrawal from chronic nicotine intoxication, which was dose specific; it occurred in rats administered 0.15 mg/kg or 0.6 mg/kg but not the 0.3 mg/kg nicotine dose. Administration of nicotine, either acutely (0.3 mg/kg) during ethanol withdrawal, or chronically (0.15, 0.3 or 0.6 mg/kg) during the chronic alcohol treatment procedure, diminished locomotor activity, which increases significantly, approximately 6-7 h after withdrawal, in rats chronically treated with alcohol. Rats which were chronically treated with alcohol alone or in combination with nicotine, 0.3 mg/kg, showed an increase in ethanol intake when the free choice was performed between ethanol 10% and tap water; on the contrary, when the free choice was performed between ethanol 10% versus nicotine, 0.3 mg/kg, results showed a decrease in ethanol preference and a concomitant increase in nicotine preference. CONCLUSION: These studies clearly identified the modulatory effects of nicotine, at specific doses, on both motility and preference in rat chronically co-administered nicotine and ethanol.     

Effects of morphine on acquisition and maintenance of ethanol and water intake patterns in rats.

Two experiments were conducted to assess the effects of chronic subcutaneous injections of morphine (1.0 mg/kg) or saline on the pattern and amount of sweetened ethanol and water intake in fluid restricted Long-Evans rats. Following daily injections, 2-h two-bottle choice tests were conducted with water and an ethanol solution (15% ethanol v/v in 5% sucrose w/v). During a 20-day acquisition phase (Experiment 1), ethanol intake patterns and amounts did not differ between saline (n = 6) and morphine (n = 6) groups. Both groups exhibited ethanol intake patterns that decreased exponentially throughout the session suggesting control by fluid restriction procedures. Morphine decreased water intake during initial periods of each session and increased intake during later periods. In Experiment 2, morphine and saline injections were reversed across three phases with the same rats. Morphine increased total ethanol consumption during the first few days of each 15-day phase, but the groups did not differ thereafter, and the initial increases produced no statistically significant group differences. Additionally, morphine augmented ethanol intake in early portions of sessions, while water intake was decreased and increased during early and later portions of each session, respectively. Analysis of the data from the last 5 days of each phase indicated that, when injected with morphine, the group which received saline during acquisition consumed significantly more ethanol solution than the group injected with morphine during acquisition. The effect on patterns of water and ethanol intake were observed, regardless of the drug injected during acquisition.(ABSTRACT TRUNCATED AT 250 WORDS)     

Assessment of GABA-B, metabotropic glutamate, and opioid receptor involvement in an animal model of binge drinking

Drinking to intoxication or binge drinking is a hallmark characteristic of alcohol abuse. Although hard to model in rodents, the scheduled high alcohol consumption (SHAC) procedure generates high, stable ethanol intake and blood ethanol concentrations in mice to levels consistent with definitions of binge drinking. The purpose of the present studies was to determine the effects of pharmacological manipulation of the opioidergic, glutamatergic, and γ-aminobutyric acid (GABA)ergic systems on binge drinking with the SHAC procedure. Parallel manipulations were conducted in mice trained in operant self-administration of either sucrose or ethanol. For the SHAC procedure, genetically heterogeneous Withdrawal Seizure Control mice were given varying periods of fluid access, with a 30-min ethanol session every third day (total of seven). Mice were pretreated intraperitoneally with naltrexone (0, 0.6, or 1.25 mg/kg), baclofen (0, 2.5, or 5.0 mg/kg), or 2-methyl-6-(phenylethynyl)-pyridine (MPEP; 0, 3.0, or 10.0 mg/kg) before each ethanol session. For the operant self-administration procedure, separate groups of C57BL/6 mice were trained to complete a single response requirement (16 presses on the active lever) to gain 30 min of access to an ethanol or a sucrose solution. Mice received pretreatments of the same doses of naltrexone, MPEP, or baclofen before the self-administration sessions, with saline injections on intervening days. Naltrexone produced a dose-dependent decrease in binge drinking, and the highest dose also significantly decreased operant self-administration of ethanol and sucrose. Both doses of baclofen significantly decreased binge alcohol consumption, but the higher dose also tended to decrease water intake. The highest dose of baclofen also significantly decreased operant self-administration of sucrose. MPEP (10 mg/kg) significantly decreased binge alcohol consumption and sucrose self-administration. These results indicate that manipulation of the opioidergic, glutamatergic, and GABAergic systems significantly decreased binge drinking.