Moderate Ethanol Consumption Research Articles

Biphasic effects of ethanol consumption on N,N-dimethylformamide-induced liver injury in mice

N,N-Dimethylformamide (DMF) is a well-documented occupational hazardous material, which can induce occupational liver injury. The current study was designed to investigate whether ethanol consumption can affect DMF-induced hepatotoxicity and the potential underlying mechanisms involved. We found that a single dose of ethanol (1.25, 2.5, or 5 g/kg bw by gavage) significantly repressed the increase in serum alanine transaminase (ALT) and aspartate transaminase (AST) activities and alleviated the liver histopathological changes in mice challenged with 3 g/kg DMF. In contrast, long-term moderate drinking (2.5 g/kg bw) significantly aggravated the repeated DMF (0.7 g/kg bw) exposure-induced increase in the serum ALT and AST activities. Mechanistically, acute ethanol consumption suppressed DMF-induced activation of the NLR family pyrin domain-containing protein 3 (NLRP3) inflammasome, while long-term moderate ethanol consumption promoted hepatocyte apoptosis in the mouse liver. Notably, cytochrome P4502E1 (CYP2E1) protein level and activity in mouse livers were not significantly affected by ethanol per se in the two models. These results confirm that regular drinking can increase the risk of DMF-induced hepatotoxicity, and suggest that DMF-handling workers should avoid consuming ethanol to reduce the risk of DMF-indued liver injury.

Differential effects of moderate chronic ethanol consumption on neurobehavior, white matter glial protein expression, and mTOR pathway signaling with adolescent brain maturation

ABSTRACT Background: Adolescent brains are highly vulnerable to heavy alcohol exposure. Increased understanding of how alcohol adversely impacts brain maturation may improve treatment outcomes. Objectives: This study characterizes short-term versus long-term effects of ethanol feeding on behavior, frontal lobe glial proteins, and mTOR signaling. Methods: Adolescent rats (8/group) were fed liquid diets containing 26% or 0% ethanol for 2 or 9 weeks, then subjected to novel object recognition (NOR) and open field (OF) tests. Frontal lobes were used for molecular assays. Results: Significant ethanol effects on OF performance occurred in the 2-week model (p < .0001). Further shifts in OF and NOR performance were unrelated to ethanol exposure in the 9-week models (p < .05 to p < .0001). Ethanol inhibited MAG1 (p < .01) and MBP (p < .0001) after 2 but not 9 weeks. However, both control and ethanol 9-week models had significantly reduced MAG1 (p < .001–0.0001), MBP (p < .0001), PDGFRA (p < .05–0.01), and PLP (p < .001–0.0001) relative to the 2-week models. GFAP was the only glial protein significantly inhibited by ethanol in both 2- (p < .01) and 9-week (p < .05) models. Concerning the mTOR pathway, ethanol reduced IRS-1 (p < .05) and globally inhibited mTOR (p < .01 or p < .001) in the 9- but not the 2-week model. Conclusions: Short-term versus long-term ethanol exposures differentially alter neurobehavioral function, glial protein expression, and signaling through IRS-1 and mTOR, which have known roles in myelination during adolescence. These findings suggest that strategies to prevent chronic alcohol-related brain pathology should consider the increased maturation-related vulnerability of adolescent brains.

β-Carotene Supplementation Improves Pancreas Function during Moderate Ethanol Consumption: Initial Characterization from a Morphological Overview.

Alcohol is believed to harm acinar cells, pancreatic ductal epithelium, and pancreatic stellate cells. After giving ethanol and/or β-carotene to C57BL/6 mice, our goal was to evaluate their biochemistry, histology, and morpho-quantitative features. There were six groups of C57BL/6 mice: 1. Group C (control), 2. Group LA (low-dose alcohol), 3. Group MA (moderate-dose alcohol), 4. Group B (β-carotene), 5. Group LA + B (low-dose alcohol combined with β-carotene), and 6. Group MA + B (moderate-dose alcohol combined with β-carotene). After the animals were euthanized on day 28, each specimen's pancreatic tissue was taken. Lipase, uric acid, and amylase were assessed using biochemical assessment. Furthermore, the examination of the pancreatic structure was conducted using Ammann's fibrosis scoring system. Finally, the morpho-quantitative characteristics of the pancreatic islets and acinar cells were determined. In the serum of the MA + B group, there were higher amounts of total amylase (825.953 ± 193.412 U/L) and lower amounts of lipase (47.139 ± 6.099 U/L) (p < 0.05). Furthermore, Ammann's fibrosis punctuation in the pancreas revealed significant variations between the groups (p < 0.001). Finally, the stereological analysis of pancreatic islets showed that the groups were different (p < 0.001). These findings suggest that antioxidant treatments might help decrease the negative effects of ethanol exposure in animal models.

Study of the relationship between the severity of alcoholic cardiomyopathy and the level of alcohol consumption in male and female rats in the model of “home drinking”

Background. Earlier, on the translational model of alcoholic cardiomyopathy (ACMP) developed by us in outbred white rats, which reproduces the main clinical diagnostic signs of this disease, it was shown that in rats of both sexes under conditions of constant 24/7 alcoholization for 24 weeks, AСMP is formed.This study purpose is a comparative assessment of the features of the ACM formation in male and female rats depending on the level of ethanol consumption in model experiments simulating "domestic drunkenness" with periodic alcoholization 24/2 for 24 weeks.Materials and methods. Experiments were performed on outbred white rats randomized into 4 groups: group 1 — control male rats (n=18), group 2 — control female rats (n = 18), group 3 — alcoholized male rats (n = 39) and group 4 — alcoholized female rats (n = 19). Control animals received a normal diet and free access to water. Animals of the 3rd and 4th groups weekly, for 24 weeks, from Friday 22.00 to Monday 8.00, received a 10 % ethanol solution as the only source of liquid with unlimited access to standard food, and the usual diet on the remaining days.Results. In animals of both sexes, after 24 weeks from the consumption start of the ethanol solution, ACM is formed, as evidenced by an increase in the end-systolic and end-diastolic sizes of the left ventricle of the heart (p = 0.0001) and a decrease in its ejection fraction (p = 0.0001), while the degree of pathological myocardial remodeling is more pronounced in females. According to cluster analysis, by the consumption of ethanol (CET), animals of both sexes are divided into 3 subgroups: low, medium and high CET, however, in males, the subgroup with an average CET prevails — 56 %, and in females with a high CET — 47 % (p = 0.0286), the low CET subgroup is minimal (16 %). At the same time, if in males in all subgroups, starting from the 8th week of alcoholization, CET dynamically decreases, then how in females in subgroups with medium and high CET, starting from the 16th week, it increases. It was found that the degree of left ventricular remodeling in females with high and moderate CET was almost 2 times higher than in males (p &lt; 0.05).Conclusion. In model experiments imitating “domestic drunkenness”, it was shown that in females the intensity of the left ventricle heart remodeling is significantly higher than in males, which, apparently, is determined by the identified gender-dependent multidirectional trends in the formation of alcoholic behavior characterized by dynamic growth. consumption of ethanol in female rats as the duration of alcoholization increases.

Abstract P2026: Moderate Ethanol Consumption Causes Right Ventricular Hypertrophy And Pulmonary Hypertension Via The Pro-inflammatory Endothelin-interleukin Pathway

The substantial increase in ethanol consumption since the Covid-19 pandemic has re-emphasized the need for extensive preclinical research on the cardiac and pulmonary effects of ethanol. Current knowledge is mostly based on ethanol effects on left ventricular (LV) function and blood pressure. Therefore, this preclinical study dealt with the effects of moderate ethanol consumption on right ventricular (RV) and pulmonary artery (PA) structure and function, and the implicated mechanisms in a pulmonary hypertension susceptible rat strain. Two groups of Sprague-Dawley (CD strain) male rats (n=7-8 each) received balanced liquid diet containing 5% ethanol (w/v) or pair-fed with isocaloric liquid diet for 8 weeks. This ethanol regimen resulted in clinically relevant blood ethanol concentration (39.8 + 1.9 mg/dL). Echocardiographic analysis showed progressive decreases in PA acceleration time/ejection time and increases in mean PA pressure (hallmarks of pulmonary hypertension) in ethanol-fed rats. Further, transthoracic echocardiography in long-axis orientation showed a significant increase in the RV internal diameter/left ventricular internal diameter during diastole (RVID-d/LVID-d), indicative of RV dilation in chronic ethanol-treated rats. Echocardiographic LV Mass/body weight and post-mortem Fulton’s index were higher in ethanol-fed rats, suggesting ethanol induced simultaneous LV and RV hypertrophy. Further, ethanol increased mean arterial pressure (measured at the end of the 8-weeks study). Ex vivo biochemical analyses showed upregulation of the vasoconstrictor/pro-inflammatory protein endothelin-1 (ET-1) and its downstream target IL-6 in the RV and lungs of ethanol-fed rats. Finally, ethanol reduced lung endothelin ETB receptor expression but had no effect on the ETA receptor, leading to higher ETA/ETB receptor ratio. Collectively, this study yields new insight into ethanol-evoked RV and pulmonary artery remodeling via the upregulation of the ET-1-IL-6 pathway.

Differential Effects of White Wine and Ethanol Consumption on Survival of Rats after a Myocardial Infarction

Studies of the cardioprotective effects of wine are mainly focused on red wines, due to their much higher content of bioactive compounds relative to white wines. Although some studies indicate a cardioprotective effect of white wine, there is no clear consensus on the existence of additional benefits of white wine over ethanol. The aim of this study was to determine and compare the effects of moderate consumption of white wine and ethanol on the survival of rats subjected to surgically induced myocardial infarction (MI). Male Sprague Dawley rats (n = 74) were randomized into three groups: water only, white wine or a 13% v/v ethanol/water solution. After a four-week drinking period, MI was induced by ligating the left anterior descending artery. The survival rate was highest in the wine group (72.2%), and lowest in the water only group (47.8%). There was no statistically significant difference in survival between the ethanol and water groups. An analysis linking drinking volumes to survival outcomes revealed that lower ethanol consumption was more prevalent in rats that survived, indicating an upper limit for the protective effects of ethanol. An opposite finding was noticed in the wine group, where no deaths occurred in rats with an average daily white wine consumption of approximately 10 mL or more. We conclude that moderate consumption of white wine has a positive effect on survival after a myocardial infarction, which cannot be attributed only to ethanol, but also to other white wine constituents.

Effects of moderate ethanol consumption as a function of n-6:n-3 dietary ratio on lipid profile, inflammation, and liver function in mice

ObjectiveIt is critical to understand how moderate ethanol exposure interacts with dietary components such as essential fatty acids to influence inflammatory processes underlying CVD pathogenesis. The purpose of this study was to examine the effects of moderate ethanol consumption and dietary n-6:n-3 fatty acid composition on markers associated with CVD in mice. MethodsTwenty-three C57BL/6J mice consumed an 18% ethanol solution or 26.9% maltose dextrin solution (isocaloric control) for 12 weeks. Within each group, the mice were fed either a high n-6 (n-6:n-3 = 50:1) diet or a balanced n-3 (n-6:n-3 = 1:1) diet ad libitum. Following the exposure period, serum samples were analyzed to assess lipid profile, inflammatory markers, antioxidant capacity, DNA damage, and liver function enzyme activity. ResultsThe control group gained more weight than the ethanol group (P = 0.020). In ethanol-exposed mice, HDL was significantly increased (P = 0.009). C-reactive protein (CRP; P < 0.001), high mobility group box 1 protein (HMGB1; P = 0.011), 8-oxo-deoxyguanosine (8-oxo-dG; P = 0.019), ALT (P = 0.002) and AP (P = 0.021) were lower in the ethanol group. There was a significant main effect of the n-3 diet on total antioxidant capacity (TAC; P < 0.001) and 8-oxo-dG (P = 0.047). ConclusionThese findings indicate that moderate ethanol consumption and a balanced n-6:n-3 diet improve several inflammatory and lipid markers associated with CVD. Observed differences in weight gain between groups should be considered when interpreting these results.

Ethanol Intoxication Impairs Respiratory Function and Bacterial Clearance and Is Associated With Neutrophil Accumulation in the Lung After Streptococcus pneumoniae Infection.

Alcohol consumption is commonplace in the United States and its prevalence has increased in recent years. Excessive alcohol use is linked to an increased risk of infections including pneumococcal pneumonia, mostly commonly caused by Streptococcus pneumoniae. In addition, pneumonia patients with prior alcohol use often require more intensive treatment and longer hospital stays due to complications of infection. The initial respiratory tract immune response to S. pneumoniae includes the production of pro-inflammatory cytokines and chemokines by resident cells in the upper and lower airways which activate and recruit leukocytes to the site of infection. However, this inflammation must be tightly regulated to avoid accumulation of toxic by-products and subsequent tissue damage. A majority of previous work on alcohol and pneumonia involve animal models utilizing high concentrations of ethanol or chronic exposure and offer conflicting results about how ethanol alters immunity to pathogens. Further, animal models often employ a high bacterial inoculum which may overwhelm the immune system and obscure results, limiting their applicability to the course of human infection. Here, we sought to determine how a more moderate ethanol exposure paradigm affects respiratory function and innate immunity in mice after intranasal infection with 104 colony forming units of S. pneumoniae. Ethanol-exposed mice displayed respiratory dysfunction and impaired bacterial clearance after infection compared to their vehicle-exposed counterparts. This altered response was associated with increased gene expression of neutrophil chemokines Cxcl1 and Cxcl2 in whole lung homogenates, elevated concentrations of circulating granulocyte-colony stimulating factor (G-CSF), and higher neutrophil numbers in the lung 24 hours after infection. Taken together, these findings suggest that even a more moderate ethanol consumption pattern can dramatically modulate the innate immune response to S. pneumoniae after only 3 days of ethanol exposure and provide insight into possible mechanisms related to the compromised respiratory immunity seen in alcohol consumers with pneumonia.

Ethanol Alleviates Amyloid-β-Induced Toxicity in an Alzheimer's Disease Model of Caenorhabiditis elegans.

Amyloid-β, a hallmark of Alzheimer’s disease, forms toxic intracellular oligomers and extracellular senile plaques resulting in neuronal toxicity. Ethanol is widely consumed worldwide. Moderate ethanol consumption has numerous benefits in humans. We found that ethanol could significantly extend the lifespan of Caenorhabiditis elegans in a previous study. Based on that study, we tested the effect of ethanol on Alzheimer’s disease transgenic Caenorhabiditis elegans strain CL4176, which expresses amyloid-β1-42 peptide in body wall muscle cells. Ethanol delayed paralysis and reduced amyloid-β oligomers in Caenorhabiditis elegans worms of the CL4176 strain. Moreover, ethanol could induce the nuclear translocation of DAF-16 in the nematodes. However, in worms that were fed daf-16 RNAi bacteria, ethanol no longer delayed the paralysis. The qPCR assays showed that ethanol increases the expression of daf-16, hsf-1 and their common target genes- small heat shock protein genes. In addition, we also found that ethanol could increase lysosome mass in the CL4176 worms. In summary, our study indicated that ethanol attenuated amyloid-β toxicity in the Alzheimer’s disease model of Caenorhabiditis elegans via increasing the level of lysosomes to promote amyloid-β degradation and upregulating the levels of small heat shock protein genes to reduce amyloid-β aggregation.

A novel murine model of multi-day moderate ethanol exposure reveals increased intestinal dysfunction and liver inflammation with age

BackgroundThere are currently > 600 million people over the age of 65 globally and this number is expected to double by the year 2050. Alcohol use among this population is on the rise, which is concerning as aging is associated with increased risk for a number of chronic illnesses. As most studies investigating the effects of alcohol have focused on young/middle-aged populations, there is a dearth of information regarding the consequences of alcohol use in older consumers. In addition, most murine ethanol models have concentrated on exposure to very high levels of ethanol, while the vast majority of elderly drinkers do not consume alcohol in excess; instead, they drink on average 2 alcoholic beverages a day, 3–4 days a week.MethodsWe designed a murine model of aging and moderate ethanol consumption to determine if the deleterious effects of alcohol on the gut-liver axis are exacerbated in aged, relative to younger, animals. Aged and young mice were exposed to a multi-day moderate exposure ethanol regimen for 4 weeks and changes in gut permeability along with intestinal tight junction protein and antimicrobial peptide gene expression were measured. In addition, hepatic inflammation was assessed by histological analysis, inflammatory gene expression and flow cytometric analysis of inflammatory infiltrate.ResultsOur results reveal that in aged, but not young mice, moderate ethanol exposure yielded significantly worsened intestinal permeability, including increased bacterial translocation from the gut, elevated serum iFABP and leakage of FITC-dextran from the gut. Interestingly, moderate ethanol exposure in young animals led to gut protective transcriptional changes in the ileum while this protective response was blunted in aged mice. Finally, moderate ethanol exposure in aged mice also resulted in marked inflammatory changes in the liver.ConclusionsThese results demonstrate that aged mice are more susceptible to ethanol-induced gut barrier dysfunction and liver inflammation, even at moderate doses of ethanol. This increased vulnerability to ethanol’s gastrointestinal effects has important implications for alcohol use in the aging population. Future studies will explore whether improving intestinal barrier function can reverse these age-related changes.

Effects of Moderate Ethanol Consumption with a n‐3 Diet on Inflammation and Total Antioxidant Capacity in Mice

Excessive alcohol intake is known to elevate risk for cardiovascular diseases (CVD); however, several studies have reported that moderate alcohol consumption may not contribute additional risk or may provide a protective effect against CVD. It is critical to understand how moderate ethanol exposure interacts with other essential dietary components such as n-3 fatty acids to influence inflammation underlying CVD pathogenesis. The purpose of this study was to examine the effects of moderate ethanol consumption as a function of dietary n-6:n-3 fatty acid composition on markers associated with inflammation and total antioxidant capacity (TAC) in mice. Twenty-three mice (12 male, 11 female) consumed an 18% ethanol solution or 26.9% maltose dextrin solution (non-ethanol isocaloric control) for 12 weeks. In each group, half of the mice were fed either a high n-6 (n-6:n-3 = 50:1) diet or a balanced n-3 (n-6:n-3 = 1:1) diet ad libitum. There were no differences in initial body weight between the two groups; however, the control group gained significantly more weight than the ethanol group (P = 0.020) associated with higher maltose dextrin fluid intake (P < 0.001). C-reactive protein and high mobility group box 1 protein were significantly lower in the ethanol group compared to the control group (P < 0.001 and P = 0.029, respectively). There was a significant main effect of n-3 diet on total antioxidant capacity (TAC; P < 0.001) and no significant effect of ethanol on TAC. These findings indicate that moderate consumption of alcohol may improve inflammatory markers associated with CVD in mice; however, the results should be interpreted with caution due to the differences in weight gain between groups.

Phosphatidylethanol in whole blood of rhesus monkeys correlates with ethanol consumption.

Phosphatidylethanol (PEth) homologs are ethanol metabolites used to identify and monitor alcohol drinking in humans. In this study, we measured levels of the 2 most abundant homologs, PEth 16:0/18:1 and PEth 16:0/18:2, in whole blood samples from rhesus macaque monkeys that drank ethanol daily ad libitum to assess the relationship between PEth levels and recent ethanol exposure in this animal model. Blood samples were obtained from The Monkey Alcohol Tissue Research Resource. The monkeys were first induced to consume 4% (w/v) ethanol in water from a panel attached to their home cage. Then, monkeys were allowed to drink ethanol and water ad libitum 22h daily for 12months and the daily amount of ethanol each monkey consumed was measured. Whole, uncoagulated blood was collected from each animal at the end of the entire experimental procedure. PEth 16:0/18:1 and PEth 16:0/18:2 levels were analyzed by HPLC with tandem mass spectrometry, and the ethanol consumed during the preceding 14days was measured. Combined PEth was the sum of the concentrations of both homologs. Our results show that (1) PEth accumulates in the blood of rhesus monkeys after ethanol consumption; (2) PEth homolog levels were correlated with the daily average ethanol intake during the 14-day period immediately preceding blood collection; (3) the application of established human PEth 16:0/18:1 cutoff concentrations indicative of light social or no ethanol consumption (<20ng/ml), moderate ethanol consumption (≥ 20 and < 200ng/ml) and heavy ethanol consumption (≥ 200ng/ml) predicted significantly different ethanol intake in these animals. PEth homologs were not detected in ethanol-naïve controls. This study confirms that PEth is a sensitive biomarker for ethanol consumption in rhesus macaque monkeys. This nonhuman primate model may prove useful in evaluating sources of variability previously shown to exist between ethanol consumption and PEth homolog levels among humans.

Chronic ethanol consumption does not reduce true bone density in male Wistar rats.

Osteoporosis is characterized by reduced bone mineral density (BMD) and increased bone fragility, which may be modified by lifestyle behaviors. In observational studies, chronic moderate ethanol consumption is associated with higher BMD, but results are inconsistent and underlying mechanisms are unknown. To understand the influence of chronic ethanol consumption on true bone density (Archimedes principal), bone mechanical properties (Young's Modulus of bend), and osteogenic gene expression, 12-month-old male Wistar rats were randomly assigned to a control group or ethanol intervention (20% ethanol in drinking water on alternate days) group for 13 weeks and tibiae and femurs were collected. Blood was collected to assess alcohol content and antioxidant enzyme activities. We hypothesized that chronic ethanol consumption would increase true bone density and mechanical properties and increase osteoblastic gene expression and serum antioxidant enzyme activity. Ethanol consumption did not influence femoral or tibial true bone density but did result in lower tibial Young's modulus of bend (p=0.0002). However, there was no influence of ethanol on other measures of mechanical properties. Femoral pro-osteoclastic gene expression of Dkk1 was lower (p=0.0006) and pro-osteoblastic gene expression of Ctnnb1 was higher (p=0.02) with ethanol consumption. We observed no differences in circulating antioxidant activities between groups, other than a tendency for greater (p=0.08) glutathione peroxidase in the ethanol group. Results showed chronic ethanol consumption did not influence true bone density, only modestly reduced tibial mechanical properties (lower Young's modulus of bend), and moderately impacted expression of genes within the femur known to regulate both osteoblast and osteoclast activities.

Potential Mechanisms of Cardiac Preconditioning : Cardioprotective Effects of Volatile Anesthetics and Chronic Mild to Moderate Ethanol Consumption

Potential Mechanisms of Cardiac Preconditioning : Cardioprotective Effects of Volatile Anesthetics and Chronic Mild to Moderate Ethanol Consumption

Low Ethanol Concentrations Promote Endothelial Progenitor Cell Capacity and Reparative Function.

Background Endothelial progenitor cells (EPCs) are recruited to injured endothelium and contribute to its regeneration. There is evidence that moderate ethanol consumption prevents the development and progression of atherosclerosis in a variety of in vitro and in vivo models and increases the mobilization of progenitor cells. Furthermore, there are studies that identified ethanol at low concentration as a therapeutic tool to mobilize progenitor cells in peripheral blood. At the same time, the cell number of EPCs represents a close link to cardiovascular system constitution and function and contributes to cardiovascular risk. The aim of this study was to evaluate the effect of low dose ethanol on typical features of endothelial colony-forming cells (ECFCs), a proliferative subtype of EPCs. Methods and Results We tested whether ethanol impacts the functional abilities of ECFC (e.g., migration, tube formation, and proliferation) using in vitro assays, the intercommunication of ECFC by exploring cell surface molecules by flow cytometry, and the expression of (anti-)angiogenic molecules by ELISA. Low concentrations of ethanol concentration promoted migration, proliferation, and tubule formation of ECFC. The expression of the cell surface marker VE-cadherin, a protein which plays an important role in cell-cell interaction, was enhanced by ethanol, while (anti-)angiogenic molecule expression was not impacted. Conclusion Ethanol at moderate concentrations increases the angiogenic abilities of endothelial progenitor cells thus possibly contributing to vasoprotection.

Effect of Moderate Intake of Ethanol on Lipid Profile of Wistar-Albino Rats Fed High Fat Diet

Many evidences suggest that light to moderate alcohol drinking may provide some cardiovascular benefits. The present study examined the impact of how moderate ethanol consumption provides a protective effect against high blood cholesterol. Wistar-Albino rats were divided into a control group (I) and test groups (II, III, IV). The test groups (II, III, IV) were administered with a predefined dosage of 10%, 20% and 30% ethanol respectively following twenty days of high fat routine however, control animals were managed with normal saline for the next fifteen days. Significant difference in lipid profile parameters witnessed before and after ethanol administration between the control and test groups. Findings from the study suggests that 10% moderate ethanol is adequately least enough to reduce the risk of cardiovascular disease by improving the blood cholesterol in Wistar-Albino rats.

Inflammation Biomarkers in Older Adult Rats Consuming Moderate Levels of Ethanol

Previous epidemiological studies and experimental data from rodent models have reported a non‐linear relationship between consumption of alcohol and cardiovascular disease risk and colorectal health. These studies indicate that light‐to‐moderate alcohol consumption, in contrast to excessive alcohol use, may provide cardiovascular health benefits as well as a protective effects on risk for colorectal cancer. Although prior studies have examined the effects of moderate ethanol consumption primarily in young adult animals, it is unknown if the results can be extrapolated to older adults or if these effects may be more pronounced in aging animals due to declining indices of health.. This study examined the effects of moderate ethanol consumption on lipid profile and colonic gene expression in older rats. Twenty‐four non deprived middle aged (420 days old) Wistar rats (n=12/group) were allowed to voluntarily consume a 20% v/v ethanol solution on alternate days for 13 weeks or were given access to water as their sole source of fluid (non‐ethanol exposed control). Moderate ethanol consumption significantly increased HDL cholesterol levels (P = 0.016) and ApoA1 (P = 0.048) compared to control. There were no differences in serum triglyceride, total cholesterol, LDL cholesterol, oxidized LDL and glucose between the groups. Blood C‐reactive protein (CRP) was decreased significantly (P = 0.034) in ethanol‐exposed animals. Blood high‐mobility group protein 1 (HMGB1) showed a decreased trend in the ethanol group (P = 0.083). In colon, gene expression of aldehyde dehydrogenase (Aldh) displayed an increased trend in ethanol‐exposed rats (P = 0.073). Colonic superoxide dismutase (Sod) was upregulated with ethanol intervention (P = 0.050). Colonic gene expression of cyclooxygenase 2 (Cox‐2), RelA, b‐catenin, CD68, and Pparr were not different between the groups. These results suggest that moderate consumption of ethanol in older rats may potentially contribute to improved cardiovascular and colorectal cancer risk by improving blood cholesterol, decreasing blood markers of inflammation in and increasing antioxidant enzyme expression in colon.Support or Funding InformationNIH AA023291

EFFECTS OF MODERATE ETHANOL CONSUMPTION ON EXPRESSION OF GENES RELATED TO COLONIC CELL GROWTH IN YOUNG ADULT AND OLDER RATS

Our previous research showed that moderate ethanol consumption reduced colonic cell proliferation regardless of age. To examine the underlying mechanism, this study examined gene expression related to colonic cell growth. Twenty‐four nondeprived young adult (109 days old) Wistar rats and twenty‐four nondeprived middle‐aged (420 days old) Wistar rats were randomly assigned to an ethanol‐exposed or a non ethanol‐exposed (water control) group (n = 12 per group). The ethanol group was provided voluntary access to a 20% v/v ethanol solution on alternate days for 13 weeks (45 ethanol drinking sessions total), a paradigm which results in mean blood alcohol levels of ~30–50 mg/dl (0.03–0.05 mg%) in the outbred Wistar strain when measured 30–120 min into a standard drinking session. The control group was provided with standard drinking water during the same period. Gene expression of cyclin D1, cyclin dependent kinase 2 (CDK2), cyclin dependent kinase 4 (CDK4), p21, E‐cadherin and p53 were determined by real time polymerase chain reaction in colonic scraped mucosa. CDK2 was downregulated in ethanol‐fed rats compared to the control group (P = 0.017). Cyclin D1 trended toward lower expression (P = 0.095), and p21 trended toward higher expression (P = 0.095) in ethanol‐fed rats compared to controls, with no observed effect of age for either gene. Older rats showed significant decreases in CDK4 (P = 0.001), E cadherin (P = 0.013), and p53 (P = 0.005), with no observed effect of ethanol exposure. These results indicate that moderate ethanol consumption may improve expression of some genes related to important parameters of colonic cell growth and tumorigenesis.Support or Funding InformationNIH AA023291

Effects of Low‐to‐Moderate Ethanol Consumption on Colonic Cell Proliferation, Differentiation and Apoptosis in Young Adult and Middle‐Aged Rats

Excessive alcohol consumption is a risk factor associated with colorectal cancer; however, some epidemiological studies have reported that moderate alcohol consumption may not contribute additional risk for developing colorectal cancer while others suggest that moderate alcohol consumption may provide a protective effect that reduces colorectal cancer risk. Prior experimental data have indicated the importance of proliferation, differentiation, and apoptosis as parameters to consider when evaluating colonic cell growth and tumorigenesis. Chemopreventative agents are reported to decrease cell proliferation and to increase differentiation and apoptosis. The present study examined whether chronic low‐to‐moderate ethanol consumption had an effect on these parameters of colonic cell growth. Twenty‐four nondeprived young adult (109 days old) Wistar rats (n=12/group) and twenty‐four nondeprived middle‐aged (420 days old) Wistar rats (n=12/group) were allowed to voluntarily consume a 20% v/v ethanol solution on alternate days for 13 weeks (45 ethanol drinking sessions total) or were given access only to water (non ethanol‐exposed control). The intermittent access 20% ethanol drinking paradigm utilized results in mean blood alcohol levels of ~30–50 mg/dl in the outbred Wistar strain when measured 30–120 min into a standard drinking session. Following the final experimental session, colon tissues were collected for quantitative immunohistochemical analyses of cell proliferation, differentiation and apoptosis using Ki‐67, goblet cell and TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labeling), respectively. Ethanol treatment resulted in a lower cell proliferation index in both young (P = 0.05) and older (P &lt; 0.001) rats, as well as a lower proliferative zone (P &lt; 0.01 for both ages). Older rats had a shorter colon, but there was no significant difference for proliferation index between young and old rats. Cell differentiation between ethanol‐treated animals and controls was not significant for young rats, but older rats had a lower level of differentiation in the ethanol group. For apoptosis, there was no significant effect of ethanol treatment for either young or old rats. Younger rats had higher values than older rats for both differentiation and apoptosis (P &lt; 0.05). These findings suggest that moderate consumption of ethanol improves at least one notable parameter (cell proliferation) in colonic tumorigenesis regardless of age, however, reduced cell differentiation among older animals.Support or Funding InformationSupport Contributed By: NIH AA023291

Effects of Moderate Ethanol Consumption with a n‐3 Diet on Lipid Profile and Liver Function in Mice

Several studies have found a non‐linear relationship between alcohol consumption and cardiovascular disease (CVD) risk, suggesting that low or moderate consumption of alcohol may provide cardioprotective effects, while excessive alcohol consumption increases the risk of CVD. Studies also suggest that increased dietary consumption of n‐3 fatty acids may reduce CVD risk. The purpose of this study is to examine the effects of moderate ethanol consumption as a function of dietary n‐6:n‐3 fatty acid composition on markers associated with cardiovascular disease and liver dysfunction in mice. Twenty‐three mice (12 male, 11 female) consumed an 18% ethanol solution or 26.9% maltose dextrin solution (non‐ethanol isocaloric control) for 12 weeks. In each group, half of the mice were fed either a high n‐6 (n‐6:n‐3 = 50:1) diet or a balanced n‐3 (n‐6:n‐3 = 1:1) diet ad libitum. There were no differences in initial body weight between the two groups; however, the control group gained significantly more weight than the ethanol group (P = 0.020) associated with higher maltose dextrin fluid intake (P &lt; 0.001). In the control group, the balanced n‐3 diet resulted in significantly lower aspartate aminotransferase (AST) levels (P = 0.010) compared to the high n‐6 diet. In the ethanol group, the balanced n‐3 diet resulted in lower levels of serum triglycerides (P = 0.086), total cholesterol (P= 0.096), and low‐density lipoprotein (LDL) levels (P = 0.053) that were approaching significance. These findings indicate that a diet with a balanced n‐6 to n‐3 ratio may improve several lipid profile markers associated with CVD in mice who concurrently consume moderate amounts of alcohol.Support or Funding InformationNIH AA023291