Prolonged Alcohol Intake Research Articles

Proanthocyanidins alleviate acute alcohol liver injury by inhibiting pyroptosis via inhibiting the ROS-MLKL-CTSB-NLRP3 pathway

BackgroundAlcoholic Liver Disease (ALD) is a hepatic disorder resulting from prolonged or excessive alcohol intake. The predominant manifestation of ALD is fatty liver, which progresses to alcoholic hepatitis as the disease worsens. Pyroptosis is a novel type of programmed cell death that is intricately linked to the inflammatory cascade, presenting a promising avenue for therapeutic intervention in the management of ALD. Oligomeric proanthocyanidins (OPCs) are polyphenols extracted from grape seeds that have anti-inflammatory and antioxidant properties. However, whether OPCs can treat ALD by suppressing pyroptosis is not completely clarified. PurposeTo explore the role of OPCs in ALD to inhibit pyroptosis and its mechanism. MethodsIn vitro, HepG2 cells were employed to evaluate the beneficial impact of OPCs on alcohol-induced pyroptosis. MTT colorimetric method, enzyme-linked immunosorbent assay (ELISA), western blot (WB), immunofluorescence, acridine orange (AO) staining, and reactive oxygen species (ROS) assay were performed. In vivo, C57BL mice were used and gavaged with alcohol and OPCs. Hematoxylin-eosin staining (HE) staining, alanine aminotransferase (ALT), aspartate aminotransferase (AST) level assay, and WB were performed. ResultsThe findings revealed that OPCs could reduce the alcohol-induced increase in pyroptosis-related proteins, such as pyrin domain-containing 3 protein (NLRP3), cleaved-caspase 1, gasdermin D (GSDMD-N), Interleukin-18 (IL-18), IL-1β (IL-1β). In in vitro mechanistic experiments, We discovered that OPCs ameliorate alcohol-induced pyroptosis by decreasing cathepsin B (CTSB) leakage-mediated NLRP3 activation. More significantly, we discovered that alcohol phosphorylates mixed lineage kinase domain-like protein (MLKL), enabling P-MLKL to translocate to the lysosomal membrane and induce lysosomal membrane permeabilization (LMP). OPCs might counteract the effects of alcohol by reducing the leakage of CTSB and inhibiting the phosphorylation of MLKL through the scavenging of ROS. Conclusions: These results suggested that OPCs might counteract ALD by inhibiting pyroptosis through the ROS-MLKL-CTSB-NLRP3 pathway. Our study offered fresh insight into the ways in which naturally occurring chemicals shield ALD against harm.

Bergenin inhibits hepatic fat deposition by activating the AMPK signaling pathway, thereby attenuating alcoholic liver disease

Alcoholic liver disease (ALD) is a prevalent liver condition that arises from prolonged and excessive alcohol intake. Bergenin (BER) is an effective phytotherapeutic agent that exhibits pharmacological properties, including anti-inflammatory and anti-oxidative effects. To establish an in vivo model of ALD, C57BL/6 mice were continuously fed a high-fat diet (HFD) and administered alcohol gavage for 8 weeks, while concurrently administering BER and evaluated for therapeutic effects. After modeling, the therapeutic effects of BER were evaluated by observing histopathological changes and the detection of relevant biochemical indicators in mice. In addition, RNA sequencing of liver tissues was performed to analyze differentially expressed genes and to investigate the associated signaling pathways in order to elucidate the protective mechanisms of BER. These differentially expressed genes were mainly enriched in lipid metabolism pathways and the cytochrome P450 metabolism of exogenous substances. Subsequently, HepG2 was co-treated with sodium oleate (NaOA) and ethanol to establish an in vitro model, and the specific mechanism by which BER ameliorates ALD was further analyzed in depth. AMPK inhibitor, Compound C (CC), was demonstrated to significantly inhibit the regulation of lipid metabolism by BER in vitro. Finally, the differentially expressed genes selected were validated through qRT-PCR and Western blot analysis. Collectively, our findings revealed that BER effectively alleviated liver injury caused by alcohol and HFD in mice, significantly suppressing lipid deposition in ALD, enhancing alcohol metabolism, and mitigating oxidative stress.

Alcoholic cardiomyopathy: an update.

Alcohol-induced cardiomyopathy (AC) is an acquired form of dilated cardiomyopathy (DCM) caused by prolonged and heavy alcohol intake in the absence of other causes. The amount of alcohol required to produce AC is generally considered as >80 g/day over 5 years, but there is still some controversy regarding this definition. This review on AC focuses on pathogenesis, which involves different mechanisms. Firstly, the direct toxic effect of ethanol promotes oxidative stress in the myocardium and activation of the renin-angiotensin system. Moreover, acetaldehyde, the best-studied metabolite of alcohol, can contribute to myocardial damage impairing actin-myosin interaction and producing mitochondrial dysfunction. Genetic factors are also involved in the pathogenesis of AC, with DCM-causing genetic variants in patients with AC, especially titin-truncating variants. These findings support a double-hit hypothesis in AC, combining genetics and environmental factors. The synergistic effect of alcohol with concomitant conditions such as hypertension or liver cirrhosis can be another contributing factor leading to AC. There are no specific cardiac signs and symptoms in AC as compared with other forms of DCM. However, natural history of AC differs from DCM and relies directly on alcohol withdrawal, as left ventricular ejection fraction recovery in abstainers is associated with an excellent prognosis. Thus, abstinence from alcohol is the most crucial step in treating AC, and specific therapies are available for this purpose. Otherwise, AC should be treated according to current guidelines of heart failure with reduced ejection fraction. Targeted therapies based on AC pathogenesis are currently being developed and could potentially improve AC treatment in the future.

Mechanism of Shenling Baizhu Powder on treatment of alcoholic liver disease by regulating TLR4/NLRP3 pathway

This study aims to investigate the regulatory effects of Shenling Baizhu Powder(SBP) on cellular autophagy in alcoholic liver disease(ALD) and its intervention effect through the TLR4/NLRP3 pathway. A rat model of chronic ALD was established by gavage of spirits. An ALD cell model was established by stimulating BRL3A cells with alcohol. High-performance liquid chromatography(HPLC) was utilized for the compositional analysis of SBP. Liver tissue from ALD rats underwent hematoxylin-eosin(HE) and oil red O staining for pathological evaluation. Enzyme-linked immunosorbent assay(ELISA) was applied to quantify lipopolysaccharides(LPS), tumor necrosis factor-alpha(TNF-α), interleukin-1 beta(IL-1β), and interleukin-18(IL-18) levels. Quantitative reverse transcription polymerase chain reaction(qRT-PCR) was conducted to evaluate the mRNA expression of myeloid differentiation factor 88(MyD88) and Toll-like receptor 4(TLR4). The effect of different drugs on BRL3A cell proliferation activity was assessed through CCK-8 analysis. Western blot analysis was performed to examine the protein expression of NOD-like receptor pyrin domain-containing 3(NLRP3), nuclear factor-kappa B P65(NF-κB P65), phosphorylated nuclear factor-kappa B P65(p-P65), caspase-1, P62, Beclin1, and microtubule-associated protein 1 light chain 3(LC3Ⅱ). The results showed that SBP effectively ameliorated hepatic lipid accumulation, reduced liver function, mitigated hepatic tissue inflammation, and reduced levels of LPS, TNF-α, IL-1β, and IL-18. Moreover, SBP exhibited the capacity to modulate hepatic autophagy induced by prolonged alcohol intake through the TLR4/NLRP3 signaling pathway. This modulation resulted in decreased expression of LC3Ⅱ and Beclin1, an elevation in P62 expression, and the promotion of autolysosome formation. These research findings imply that SBP can substantially enhance liver function and mitigate lipid irregularities in the context of chronic ALD. It achieves this by regulating excessive autophagic responses caused by prolonged spirit consumption, primarily through the inhibition of the TLR4/NLRP3 pathway.

Nutritional ketosis as treatment for alcohol withdrawal symptoms in female C57BL/6J mice

Upon both acute and prolonged alcohol intake, the brain undergoes a metabolic shift associated with increased acetate metabolism and reduced glucose metabolism, which persists during abstinence, putatively leading to energy depletion in the brain. This study evaluates the efficacy of ketogenic treatments to rescue psychiatric and neurochemical alterations during long-term alcohol withdrawal. Female mice were intermittently exposed to alcohol vapor or air for three weeks, during which mice were introduced to either a ketogenic diet (KD), control diet supplemented with ketone ester (KE) or remained on control diet (CD). Withdrawal symptoms were assessed over a period of four weeks followed by re-exposure using several behavioral and biochemical tests. Alcohol-exposed mice fed CD displayed long-lasting depressive-like symptoms measured by saccharin preference and tail suspension, as well as decreased norepinephrine levels and serotonin turnover in the hippocampus. Both KD and KE rescued anhedonia for up to three weeks of abstinence. KD mice showed higher latency to first immobility in the tail suspension test, as well as lower plasma cholesterol levels. Our findings show promising effects of nutritional ketosis in ameliorating alcohol withdrawal symptoms in mice. KD seemed to better rescue these symptoms compared to KE.

Effects of Acetyl-L-Carnitine Administration on Auditory Evoked Potentials in Rats Exposed to Chronic Ethanol

These days, one of the biggest issues facing public health is chronic alcohol consumption. Prolonged alcohol intake has been associated with a number of illnesses, including pancreatitis, cerebrovascular illnesses, and liver damage. It is known that the endogenous molecule acetyl-L-carnitine (ALCAR) has neuroprotective properties. The objective of this study was to examine the impact of ALCAR on auditory evoked potentials (AEP) in a model of chronic alcoholism in rats. Four groups (n = 10) were randomly formed from 40 three-month-old albino Wistar rats: Sham (Sh), chronic ethanol (CE), ALCAR administered (ALCAR), and chronic ethanol+ALCAR (CE+ALCAR). The rats were administered either distilled water, ethanol, ALCAR, or ethanol+ALCAR via gavage for a duration of 4 weeks. AEP recordings were acquired from each rat upon the conclusion of the experiment period. The amplitudes of P1N1 and N1P2, in addition to the latencies of the P1, N1, and P2 peaks, were analyzed. Our findings showed that P1, N1, and P2 latencies were considerably prolonged in the CE group compared to the Sh group. Furthermore, the amplitudes of P1N1 and N1P2 exhibited an increase in the CE group relative to the Sh group, a decline in the CE+ALCAR group in comparison to the CE group, and a subsequent return to normal levels. Finally, our results demonstrated that, at the dose and duration used here, ALCAR normalized the increased neuronal activity but had no effect on the conduction velocity of the hearing signal.

Capturing Russian drinking patterns with the Alcohol Use Disorders Identification Test: An exploratory interview study in primary healthcare and narcology centers in Moscow.

Despite a considerable reduction in alcohol consumption, Russia has one of the highest levels of alcohol-attributable burden of disease worldwide due to heavy episodic drinking patterns. Further improvement of alcohol control measures, including early provision of screening and brief interventions (SBI), is needed. The legislative framework for delivering SBI in Russia was introduced in 2013. As part of the creation and validation of a Russian version of the Alcohol Use Disorders Identification Test (AUDIT), the present contribution explored challenges in using the AUDIT in Russia to inform a subsequent validation study of the tool. Qualitative in-depth expert interviews with patients and healthcare professionals from four primary healthcare and narcology facilities in Moscow. A total of 25 patients were interviewed, 9 from a preventive medicine hospital, 8 from a polyclinic, and 9 from narcology clinics. Also, 12 healthcare professionals were interviewed, 5 of whom were primary healthcare doctors and 7 were narcologists. Patients and healthcare professionals expressed difficulties in dealing with the concept of a "standard drink" in the AUDIT, which is not used in Russia. Various patients struggled with understanding the meaning of "one drinking occasion" on the test, mainly because Russian drinking patterns center around festivities and special occasions with prolonged alcohol intake. Narcology patients had specific difficulties because many of them experienced zapoi-a dynamic drinking pattern with heavy use and a withdrawal from social life, followed by prolonged periods of abstinence. Surrogate alcohol use was described as a common marker of alcohol dependence in Russia, not accounted for in the AUDIT. The provided analyses on the perception of the Russian AUDIT in different patient and professional groups suggest that a series of amendments in the test should be considered to capture the specific drinking pattern and its potential harms.

Effects of Alcohol Consumption on Various Systems of the Human Body: A Systematic Review.

Prolonged alcohol intake for many years has been known to cause serious ailments in human beings since time memorial. Even after knowing that this dangerous addiction paves the way to one's own grave, there isn't much difference in the way the community sees this deadly habit. Time and again history has proven that this fatal addiction could make the life of those who consume it terrible. Also, the lives of the dear ones of alcoholic people are affected as alcohol not only affects those who consume them but also kin and friends. Various research studies conducted over many years clearly show the association of prolonged alcohol intake in the causation, aggravation, worsening, and deterioration of the health of its consumers. Moreover, chronic alcohol intake single-handedly is one of the major etiological factors in various serious diseases.

Alcohol Use Disorder, Neurodegeneration, Alzheimer's and Parkinson's Disease: Interplay Between Oxidative Stress, Neuroimmune Response and Excitotoxicity.

Alcohol use disorder (AUD) has been associated with neurodegenerative diseases such as Alzheimer’s and Parkinson’s disease. Prolonged excessive alcohol intake contributes to increased production of reactive oxygen species that triggers neuroimmune response and cellular apoptosis and necrosis via lipid peroxidation, mitochondrial, protein or DNA damage. Long term binge alcohol consumption also upregulates glutamate receptors, glucocorticoids and reduces reuptake of glutamate in the central nervous system, resulting in glutamate excitotoxicity, and eventually mitochondrial injury and cell death. In this review, we delineate the following principles in alcohol-induced neurodegeneration: (1) alcohol-induced oxidative stress, (2) neuroimmune response toward increased oxidants and lipopolysaccharide, (3) glutamate excitotoxicity and cell injury, and (4) interplay between oxidative stress, neuroimmune response and excitotoxicity leading to neurodegeneration and (5) potential chronic alcohol intake-induced development of neurodegenerative diseases, including Alzheimer’s and Parkinson’s disease.

Novel Biomarker for the Diagnosis and Prognosis of Acute Alcoholic Hepatitis.

Alcohol use disorder (AUD) characterized by heavy and prolonged alcohol intake could cause various forms of alcoholic liver disease (ALD). The spectrum of alcoholic liver disease (ALD) consists of steatosis, steatohepatitis, cirrhosis and hepatocellular carcinoma depending on the duration and severity of alcohol intake [1,2]. Interestingly only one-third of heavy drinkers (typically a sub-group of AUD patients), who exhibit specific heavy drinking patterns [3,4] would develop any clinically relevant form of liver damage.

Assessment of Bone Mineral Density in Nonalcoholic Steatohepatitis Cirrhosis

Background: Cirrhosis is characterized by diffused hepatic fibrosis and nodule formation that can occur at any age. It has significant morbidity and mortality. Worldwide common causes of cirrhosis are viral hepatitis (hepatitis B virus and hepatitis C virus), prolonged excessive alcohol intake and nonalcoholic fatty liver disease (NAFLD). Progression of chronic liver disease (CLD) and deterioration of liver function are associated with various hepatic complications. Hepatic osteodystrophy is an important extrahepatic manifestation of advanced liver disease with features of classical osteoporosis and an increased risk for fractures. The objective of the study to assess the bone mineral density (BMD) by duel energy x-ray absorptiometry (DEXA) in patients with nonalcoholic steatohepatitis (NASH) cirrhosis of liver.
 Patients and Methods: A cross sectional observational study was carried out at National Institute of Nuclear Medicine and Allied Sciences (NINMAS), Bangabandhu Sheikh Mujib Medical University (BSMMU) campus, Dhaka, from July, 2016 to June, 2017. This study included the diagnosed cases of NASH cirrhosis patients presented at outpatient or inpatient department of Hepatology of BSMMU. For measurement of the BMD dual energy x-ray absorptiometry (DEXA) scan was performed by DMS Strator DR Bone densitometer at NINMAS. BMD were measured at lumbar vertebrae L1-L4 and both the femoral neck. Analysis of data from DEXA was computerized and completely automated (software 3DXA, Medix DR.) 
 Results: A total of 54 participants was included and among them 27 were cirrhotic patients (cirrhotic group) & 27 were non-cirrhotic patients with healthy liver (non-cirrhotic group). The age range of the patients was 40-70 years in both groups. Male to female ratio was 1:1.45. The mean age was 56.3 ± 6.7 years in cirrhotic group and 55.0 ± 6.3 years in non cirrhotic group. In cirrhotic group, 22 (66.7%) patients had low BMD and five (23.8%) patients had normal BMD. In non-cirrhotic group, 11 (33.3%) patients had low BMD and 16 (76.2%) patients had normal BMD. The difference was statistically significant (p < 0.05) between two groups. Multivariate logistic regression analysis was done to see the effect of multiple independent variables (age, menopause and cirrhosis) on dependent variable (low BMD). In cirrhotic patients odds ratio (OR) was 1.961 (95% CI 0.110 to 25.893) which implies cirrhotic patients had 1.961 times greater chance to develop low BMD than non cirrhotic patients keeping all other factors in fixed level.
 Conclusion: Hepatic osteodystrophy is an important extrahepatic complication of cirrhosis. NASH cirrhosis is found among elderly people and female. Osteoporosis is common in aged people and menopausal female. But in this study it was observed that a NASH cirrhosis patient is highly significant risk factor for low BMD (osteopenia and osteoporosis) other than aged persons and menopausal females. So, special measures and monitoring should be taken regarding osteoporosis in NASH cirrhotic patient to reduce subsequent morbidity.
 Bangladesh J. Nuclear Med. 21(1): 16-20, January 2018

Early post-natal neuroactive steroid manipulation modulates ondansetron effects on initial periods of alcohol consumption in rats.

Neuroactive steroids (NS) such as allopregnanolone are crucial for brain development and adult behaviour. Early post-natal alterations of NS by administering finasteride induce a decrease in the sensitivity to stimulant effects of low alcohol doses, an increase in alcohol consumption, and a decrease in ventrostriatal dopamine and serotonin levels. The aim of the present study is to observe if the effects of the 5HT3 receptor antagonist ondansetron on initial alcohol consumption are modulated by post-natal NS manipulation. For this purpose, allopregnanolone, finasteride, or vehicle was injected from day 5 to 9. In adulthood, a novel object preference test was carried out in order to detect a possible novelty-seeking pattern in our animals, which has been related to vulnerability to drug abuse. The subjects then had access to two bottles (alcohol or control solutions) one hour daily for two consecutive weeks. Ondansetron (0.01 mg/kg, 0.1 mg/kg or vehicle) was administered before the hour of consumption in the initial phase (days 1, 2, 3) of the procedure, and after prolonged alcohol intake (days 11, 12, 13). Results indicated that finasteride animals showed a higher preference to explore the new object, as well as a higher alcohol consumption than the rest of the groups. Moreover, 0.1 mg/kg of ondansetron decreased alcohol consumption, but only in the post-natal finasteride group, suggesting a possible increase in 5HT3 receptor sensitivity in these animals. In conclusion, NS manipulation in crucial stages of development, such as early post-natal periods, seems to play an important role on the effects of ondansetron on alcohol intake and in the vulnerability to develop drug use or abuse.

Effects of chronic alcohol consumption, withdrawal and nerve growth factor on neuropeptide Y expression and cholinergic innervation of the rat dentate hilus

Several studies have demonstrated the vulnerability of the hippocampal formation (HF) to chronic alcohol consumption and withdrawal. Among the brain systems that appear to be particularly vulnerable to the effects of these conditions are the neuropeptide Y (NPY)-ergic and the cholinergic systems. Because these two systems seem to closely interact in the HF, we sought to study the effects of chronic alcohol consumption (6months) and subsequent withdrawal (2months) on the expression of NPY and on the cholinergic innervation of the rat dentate hilus. As such, we have estimated the areal density and the somatic volume of NPY-immunoreactive neurons, and the density of the cholinergic varicosities. In addition, because alcohol consumption and withdrawal are associated with impaired nerve growth factor (NGF) trophic support and the administration of exogenous NGF alters the effects of those conditions on various cholinergic markers, we have also estimated the same morphological parameters in withdrawn rats infused intracerebroventricularly with NGF. NPY expression increased after withdrawal and returned to control values after NGF treatment. Conversely, the somatic volume of these neurons did not differ among all groups. On other hand, the expression of vesicular acetylcholine transporter (VAChT) was reduced by 24% in ethanol-treated rats and by 46% in withdrawn rats. The administration of NGF to withdrawn rats increased the VAChT expression to values above control levels. These results show that the effects of prolonged alcohol intake and protracted withdrawal on the hilar NPY expression differ from those induced by shorter exposures to ethanol and by abrupt withdrawal. They also suggest that the normalizing effect of NGF on NPY expression might rely on the NGF-induced improvement of cholinergic neurotransmission in the dentate hilus.

Alcohol-induced changes in opioid peptide levels in adolescent rats are dependent on housing conditions.

BackgroundEndogenous opioids are implicated in the mechanism of action of alcohol and alcohol affects opioids in a number of brain areas, although little is known about alcohol's effects on opioids in the adolescent brain. One concern, in particular when studying young animals, is that alcohol intake models often are based on single housing that may result in alcohol effects confounded by the lack of social interactions. The aim of this study was to investigate short- and long-term alcohol effects on opioids and the influence of housing conditions on these effects.MethodsIn the first part, opioid peptide levels were measured after one 24-hour session of single housing and 2-hour voluntary alcohol intake in adolescent and adult rats. In the second part, a model with a cage divider inserted during 2-hour drinking sessions was tested and the effects on opioids were examined after 6 weeks of adolescent voluntary intake in single-and pair-housed rats, respectively.ResultsThe effects of single housing were age specific and affected Met-enkephalin-Arg6Phe7 (MEAP) in particular. In adolescent rats, it was difficult to distinguish between effects induced by alcohol and single housing, whereas alcohol-specific effects were seen in dynorphin B (DYNB), beta-endorphin (BEND), and MEAP levels in adults. Voluntary drinking affected several brain areas and the majority of alcohol-induced effects were not dependent on housing. However, alcohol effects on DYNB and BEND in the amygdala were dependent on housing. Housing alone affected MEAP in the cingulate cortex.ConclusionsAge-specific housing- and alcohol-induced effects on opioids were found. In addition, prolonged voluntary alcohol intake under different housing conditions produced several alcohol-induced effects independent of housing. However, housing-dependent effects were found in areas implicated in stress, emotionality, and alcohol use disorder. Housing condition and age may therefore affect the reasons and underlying mechanisms for drinking and could potentially affect the outcome of a number of end points in research on alcohol intake.

Alcohol Induces Sensitization to Gluten in Genetically Susceptible Individuals: A Case Control Study

BackgroundThe mechanisms of cerebellar degeneration attributed to prolonged and excessive alcohol intake remain unclear. Additional or even alternative causes of cerebellar degeneration are often overlooked in suspected cases of alcohol-related ataxia. The objectives of this study were two fold: (1) to investigate the prevalence of gluten-related serological markers in patients with alcohol-related ataxia and; (2) to compare the pattern of brain involvement on magnetic resonance imaging between patients with alcohol and gluten ataxias.Materials & MethodsPatients diagnosed with alcohol and gluten ataxias were identified from a retrospective review of patients attending a tertiary clinic. HLA genotype and serological markers of gluten-related disorders were recorded. Cerebellar volumetry, MR spectroscopy and voxel-based morphometric analyses were performed on patients and compared with matched control data.ResultsOf 904 registered patients, 104 had alcohol ataxia and 159 had gluten ataxia. 61% of the alcohol ataxia group and 70% of the gluten ataxia group had HLA DQ2/DQ8 genotype compared to 30% in healthy local blood donors. 44% of patients with alcohol ataxia had antigliadin antibodies compared to 12% in the healthy local population and 10% in patients with genetically confirmed ataxias. None of the patients with alcohol ataxia and antigliadin antibodies had celiac disease compared to 40% in patients with gluten ataxia. The pattern of structural brain abnormality in patients with alcohol ataxia who had antigliadin antibodies differed from gluten ataxia and was identical to that of alcohol ataxia.ConclusionsAlcohol related cerebellar degeneration may, in genetically susceptible individuals, induce sensitization to gluten. Such sensitization may result from a primary cerebellar insult, but a more systemic effect is also possible. The duration and amount of exposure to alcohol may not be the only factors responsible for the cerebellar insult.

Inflexible and Indifferent Alcohol Drinking in Male Mice

Alcoholism is characterized by compulsive alcohol intake, but this critical feature of alcoholism is seldom captured in preclinical studies. Here, we evaluated whether alcohol-preferring C57BL/6J mice develop compulsive alcohol drinking patterns, using adulteration of the alcohol solution with quinine, in a limited access choice paradigm. We assessed 2 independent aspects of compulsive drinking: (i) inflexible alcohol intake by testing whether mice would drink bitter alcohol solutions if this was their only source of alcohol and (ii) indifferent drinking by comparing intake of aversive and nonaversive alcohol solutions. Male C57BL/6J mice consumed alcohol for 2 or 8 consecutive weeks. The alcohol solution was then adulterated with graded quinine concentrations, and the effect on alcohol intake was determined. C57BL/6J mice rapidly developed compulsive alcohol drinking patterns. Adulteration of the alcohol solution with an aversive quinine concentration failed to reduce intake, indicative of inflexible drinking behavior, after only 2 weeks of alcohol experience, although quinine adulteration did suppress the acquisition of alcohol drinking in naïve mice. After 8 weeks of alcohol consumption, the mice also became indifferent to quinine. They consumed an aversive, quinine-containing alcohol solution, despite the simultaneous availability of an unadulterated alcohol solution. Prolonged alcohol ingestion did not alter the sensitivity to the bitter taste of quinine itself. These findings demonstrate the staged occurrence in mice of 2 distinct behavioral characteristics of alcoholism, i.e., inflexible and indifferent alcohol drinking.

Moderate alcohol consumption aggravates high-fat diet induced steatohepatitis in rats

The detrimental effects of chronic and excessive alcohol consumption, e.g., prolonged alcohol intake in excess of approximately 100 g/day, on human health have been well documented (Bloss, 2005; Dawson et al., 2005; Ellison et al., 2001; Hoek and Pastorino, 2002; Seitz and Stickel, 2007; Wang, 2005). Recently, a growing number of evidence shows that “moderate” alcohol consumption without heavy drinking episodes is associated with multiple health benefits and lower risks for some chronic diseases (Koppes et al., 2005; Rimm et al., 1991; Stampfer et al., 2005). The term “moderate alcohol” has been established as 1~2 drinks/day for men (Dawson, 2003), which is equivalent to 14–28 g/day based on the energy value of pure ethanol (Meister et al., 2000). Data from the Framingham Heart Study revealed that the age-adjusted mean alcohol consumption was 21.0~30.6 g/day among adult men and 10.4~14.2 g/day among adult women (Zhang et al., 2008). The inferior limit for the moderate alcohol consumption category is established on 4 drinks/day (Dawson et al., 1995), which accounts for 19.6% of a total 2000 kcal intake. Nonalcoholic steatohepatitis (NASH) has been recognized as a severe stage of nonalcoholic fatty liver disease, the most common form of chronic liver disease in the United States (Ong and Younossi, 2007). Diagnosis of NASH is defined by the concurrence of fat accumulation and infiltration of inflammatory cells in the liver, in the absence of chronic & excessive alcohol consumption history (Brunt, 2005). Certain risk factors for NASH development have been suggested to potentiate the capacity of alcohol-mediated liver damage (Diehl, 2004). Although patients with NASH have been advised against drinking alcohol, experimental evidence is lacking regarding whether alcohol consumption, especially at moderate levels, can increase their susceptibility to more liver injuries. Previously, a high-fat diet induced NASH rat model was developed by feeding rats a 71% liquid high-fat diet enriched with corn oil for three weeks (Lieber et al., 2004). Recently, we demonstrated that a higher rate of cellular apoptosis was induced in this rat model, which was associated with increased hepatic inflammation and oxidative stress (Wang et al., 2008). Since excessive alcohol feeding to rats increased apoptosis (Baroni et al., 1994; Mi et al., 2000; Slomiany et al., 1999), we investigated whether moderate alcohol consumption (16% of total calories intake) with the pre-existing NASH condition, aggravates apoptosis and hepatic inflammation in rats.

The anxiolytic etifoxine protects against convulsant and anxiogenic aspects of the alcohol withdrawal syndrome in mice

Change in the function of γ-aminobutyric acid A (GABA A) receptors attributable to alterations in receptor subunit composition is one of main molecular mechanisms with those affecting the glutamatergic system which accompany prolonged alcohol (ethanol) intake. These changes explain in part the central nervous system hyperexcitability consequently to ethanol administration cessation. Hyperexcitability associated with ethanol withdrawal is expressed by physical signs, such as tremors, convulsions, and heightened anxiety in animal models as well as in humans. The present work investigated the effects of anxiolytic compound etifoxine on ethanol-withdrawal paradigms in a mouse model. The benzodiazepine diazepam was chosen as reference compound. Ethanol was given to NMRI mice by a liquid diet at 3% for 8 days, then at 4% for 7 days. Under these conditions, ethanol blood level ranged between 0.5 and 2 g/L for a daily ethanol intake varying from 24 to 30 g/kg. These parameters permitted the emergence of ethanol-withdrawal symptoms once ethanol administration was terminated. Etifoxine (12.5–25 mg/kg) and diazepam (1–4 mg/kg) injected intraperitoneally 3 h 30 min after ethanol removal, decreased the severity in handling-induced tremors and convulsions in the period of 4–6 h after withdrawal from chronic ethanol treatment. In addition when administered at 30 and 15 min, respectively, before the light and dark box test, etifoxine (50 mg/kg) and diazepam (1 mg/kg) inhibited enhanced aversive response 8 h after ethanol withdrawal. Etifoxine at 25 and 50 mg/kg doses was without effects on spontaneous locomotor activity and did not exhibit ataxic effects on the rota rod in animals not treated with ethanol. These findings demonstrate that the GABAergic compound etifoxine selectively reduces the physical signs and anxiety-like behavior associated with ethanol withdrawal in a mouse model and may hold promise in the treatment of ethanol-withdrawal syndrome in humans.

Cross-Talk Between Pulmonary Injury, Oxidant Stress, and Gap Junctional Communication

Gap junction channels interconnect several different types of cells in the lung, ranging from the alveolar epithelium to the pulmonary vasculature, each of which expresses a unique subset of gap junction proteins (connexins). Major lung functions regulated by gap junctional communication include coordination of ciliary beat frequency and inflammation. Gap junctions help enable the alveolus to regulate surfactant secretion as an integrated system, in which type I cells act as mechanical sensors that transmit calcium transients to type II cells. Thus, disruption of epithelial gap junctional communication, particularly during acute lung injury, can interfere with these processes and increase the severity of injury. Consistent with this, connexin expression is altered during lung injury, and connexin-deficiency has a negative impact on the injury response and lung-growth control. It has recently been shown that alcohol abuse is a significant risk factor associated with acute respiratory distress syndrome. Oxidant stress and hormone-signaling cascades in the lung induced by prolonged alcohol ingestion are discussed, as well as the effects of these pathways on connexin expression and function.

Skeletal muscle changes in chronic alcoholic patients. A conventional, histochemical, ultrastructural and morphometric study.

Muscle biopsies of 20 alcoholic patients (15 males and 5 females), most of whom had liver disease and with no clinical or analytical evidence of neuromyopathy, were studied. 10 abstemious patients with no neuromuscular disease were selected as controls. Conventional histology failed to show significant skeletal muscle changes. A characteristic histochemical picture of tubular aggregates was found in one patient. An enlargement of the intermyofibrillar space with increase in glycogen deposition and fat droplets were detected ultrastructurally while 2 of the patients showed tubular aggregates. The mitochondriae of the alcoholic patients had a smaller perimeter and area than those of the control group (P less than 0.004 and P less than 0.008, respectively). These results suggest that the prolonged ingestion of alcohol can cause a mitochondrial alteration only evidenced by morphometry, with poor clinical, biochemical, electrophysiological and histological expression of myopathy. These findings can represent an early stage of alcoholic skeletal muscle injury.