Effects Of Chronic Ethanol Consumption Research Articles

Chronic binge drinking-induced susceptibility to colonic inflammation is microbiome-dependent

ABSTRACT Alterations in intestinal permeability and the gut microbiome caused by alcohol abuse are associated with alcoholic liver disease and with worsening of inflammatory bowel diseases (IBD) symptoms. To resolve the direct effects of chronic ethanol consumption on the colon and its microbiome in the absence of acute or chronic alcohol-induced liver disease, we developed a mouse model of chronic binge drinking that uncovers how alcohol may enhance susceptibility to colitis via the microbiota. Employing daily ethanol gavage, we recapitulate key features of binge ethanol consumption. We found that binge ethanol drinking worsens intestinal infection, colonic injury and inflammation, and this effect persists beyond the drinking period. Using gnotobiotics, we showed that alcohol-driven susceptibility to colitis is microbiota-dependent and transferable to ethanol-naïve mice by microbiome transplantation. Allobaculum spp. expanded in binge drinking mice, and administration of Allobaculum fili was sufficient to enhance colitis in non-drinking mice. Our study provides a model to study binge drinking-microbiota interactions and their effects on host disease and reinforces the pathogenic function of Allobaculum spp. as colitogenic bacteria. Our findings illustrate how chronic binge drinking-induced alterations of the microbiome may affect susceptibility to IBD onset or flares.

Effect of Chronic Ethanol Consumption on Exogenous Glucose Metabolism in Rats Using [1-13C], [2-13C], and [3-13C]glucose Breath Tests.

The C3 carbon of glucose molecules becomes the C1 carbon of pyruvate molecules during glycolysis, and the C1 and C2 carbons of glucose molecules are metabolized in the tricarboxylic acid (TCA) cycle. Utilizing this position-dependent metabolism of C atoms in glucose molecules, [1-13C], [2-13C], and [3-13C]glucose breath tests are used to evaluate glucose metabolism. However, the effects of chronic ethanol consumption remain incompletely understood. Therefore, we evaluated glucose metabolism in ethanol-fed rats using [1-13C], [2-13C], and [3-13C]glucose breath tests. Ethanol-fed (ERs) and control rats (CRs) (n = 8 each) were used in this study, and ERs were prepared by replacing drinking water with a 16% ethanol solution. We administered 100 mg/kg of [1-13C], [2-13C], or [3-13C]glucose to rats and collected expired air (at 10-min intervals for 180 min). We compared the 13CO2 levels (Δ13CO2, ‰) of breath measured by IR isotope ratio spectrometry and area under the curve (AUC) values of the 13CO2 levels-time curve between ERs and CRs. 13CO2 levels and AUCs after administration of [1-13C]glucose and [2-13C]glucose were lower in ERs than in CRs. Conversely, the AUC for the [3-13C]glucose breath test showed no significant differences between ERs and CRs, although 13CO2 levels during the 110-120 min interval were significantly high in ERs. These findings indicate that chronic ethanol consumption diminishes glucose oxidation without concomitantly reducing glycolysis. Our study demonstrates the utility of 13C-labeled glucose breath tests as noninvasive and repeatable methods for evaluating glucose metabolism in various subjects, including those with alcoholism or diabetes.

Effects of a Novel Beta Lactam Compound, MC-100093, on the Expression of Glutamate Transporters/Receptors and Ethanol Drinking Behavior of Alcohol-Preferring Rats.

Chronic ethanol exposure affects the glutamatergic system in several brain reward regions including the nucleus accumbens (NAc). Our laboratory has shown that chronic exposure to ethanol reduced the expression of glutamate transporter 1 (GLT-1) and cystine/glutamate exchanger (xCT) and, as a result, increased extracellular glutamate concentrations in the NAc of alcohol-preferring (P) rats. Moreover, previous studies from our laboratory reported that chronic ethanol intake altered the expression of certain metabotropic glutamate receptors in the brain. In addition to central effects, chronic ethanol consumption induced liver injury, which is associated with steatohepatitis. In the present study, we investigated the effects of chronic ethanol consumption in the brain and liver. Male P rats had access to a free choice of ethanol and water bottles for five weeks. Chronic ethanol consumption reduced GLT-1 and xCT expression in the NAc shell but not in the NAc core. Furthermore, chronic ethanol consumption increased fat droplet content as well as peroxisome proliferator-activated receptor alpha (PPAR-α) and GLT-1 expression in the liver. Importantly, treatment with the novel beta-lactam compound, MC-100093, reduced ethanol drinking behavior and normalized the levels of GLT-1 and xCT expression in the NAc shell as well as normalized GLT-1 and PPAR-α expression in the liver. In addition, MC-100093 attenuated ethanol-induced increases in fat droplet content in the liver. These findings suggest that MC-100093 may be a potential lead compound to attenuate ethanol-induced dysfunction in the glutamatergic system and liver injury. SIGNIFICANCE STATEMENT: This study identified a novel beta-lactam, MC-100093, that has demonstrated upregulatory effects on GLT-1. MC-100093 reduced ethanol drinking behavior and normalized levels of GLT-1 and xCT expression in the NAc shell as well as normalized GLT-1 and PPAR-α expression in the liver. In addition, MC-100093 attenuated ethanol-induced increases in fat droplet content in the liver.

ENDURANCE AND INTOXICATION INDICATORS OF RAT BRAIN WITH CHRONIC ALCOHOLIZATION

Introduction. One of the current important and unsolved problems of modern medicine is revealing the nature of the impact of chronic alcohol intoxication on functional indicators of life. It is important to analyze the metabolic shifts caused by ethanol, as well as to findeffectiveways to prevent and correct the effectsof alcohol. Aims. Research the effectof chronic ethanol consumption on endurance of rats, and determination of indicators of inflammation,intoxication and nonspecificresistance in the brain of laboratory rats. Materials and Methods. The experiment was performed on 2-month-old male and female rats. Alcohol intoxication was reproduced by introducing ethanol into drinking water, starting with 5% and gradually increasing to 15%. Intact groups of males and females received water. Endurance of rats was assessed once a week by the Porsolta Behavioral Despair Test, in which rats were given negative buoyancy by additional weight. The time of endurance of rats served as an indicator of fatigue. The duration of the experiment constituted 108 days. Inflammation(elastase, acid phosphatase activity), intoxication (urease activity and malonic dialdehyde activity), nonspecificresistance (lysozyme and catalase activity) were determined in the brain. Main results. It was found that the exercise stress contributes to the formation of more pronounced endurance of females than males. Chronic administration of ethanol did not significantlyaffectthe endurance of males during the swimming test, and led to a decrease in endurance of females. Biochemical studies showed that prolonged administration of ethanol to male and female rats leads to the accumulation of toxic products (urease, ammonia, lipid peroxidation products), stimulates the intensificationof inflammation(increased activity of acid phosphatase) against the suppression of nonspecificresistance (decrease of catalase and lysozyme activity) in the brain of rats under chronic alcohol intoxication. Conclusions. Alcohol consumption did not significantlyaffet the endurance of males, but reduced the endurance of females. The survey of the brain of rats showed the accumulation of toxic products, the intensificationof inflammationagainst the background of the suppression of nonspecificresistance under the influenceof ethanol.

Bone impairment in adolescent female rats chronically exposed to ethanol

Ethanol consumption has increased among teenagers worldwide considerably,including females. Long-term ethanol consumption in women has been reported to cause bone metabolism imbalance. However, only few studies are available on the impact of long-term ethanol consumptionon bone morphology during adolescence. Here, we report the effects of chronic ethanol consumption on bone structure in adolescent female rats. Twenty female Wistar rats (35 days old) received, by gavage, distilled water (control) or ethanol (6.5 g/kg/day, 22.5% w/v) once daily for 55 days. After ethanol administration, animals were perfused, and the femora were collected. Morphometric evaluations were performed by electron microscopy scanning. Femora length, cortical bone thickness and medullar bone diameter was measured. The results demonstrated that ethanol exposure during adolescence reduced the length of femurs, with a decrease of the anterior thickness, posterior thickness, and mid-lateral diameter (P<0.05). Thus, long-term ethanol intake may lead to alterations on bone morphometry, reducing the thickness of compact bone and femur length in adolescent females.

Transcriptional analysis of the response of C. elegans to ethanol exposure

Ethanol-induced transcriptional changes underlie important physiological responses to ethanol that are likely to contribute to the addictive properties of the drug. We examined the transcriptional responses of Caenorhabditis elegans across a timecourse of ethanol exposure, between 30 min and 8 h, to determine what genes and genetic pathways are regulated in response to ethanol in this model. We found that short exposures to ethanol (up to 2 h) induced expression of metabolic enzymes involved in metabolizing ethanol and retinol, while longer exposure (8 h) had much more profound effects on the transcriptome. Several genes that are known to be involved in the physiological response to ethanol, including direct ethanol targets, were regulated at 8 h of exposure. This longer exposure to ethanol also resulted in the regulation of genes involved in cilia function, which is consistent with an important role for the effects of ethanol on cilia in the deleterious effects of chronic ethanol consumption in humans. Finally, we found that food deprivation for an 8-h period induced gene expression changes that were somewhat ameliorated by the presence of ethanol, supporting previous observations that worms can use ethanol as a calorie source.

Effect of chronic ethanol consumption in rhesus macaques on the nucleus accumbens core transcriptome.

The nucleus accumbens core (NAcc) has been repeatedly demonstrated to be a key component of the circuitry associated with excessive ethanol consumption. Previous studies have illustrated that in a nonhuman primate (NHP) model of chronic ethanol consumption, there is significant epigenetic remodeling of the NAcc. In the current study, RNA-Seq was used to examine genome-wide gene expression in eight each of control, low/binge (LD*), and high/very high (HD*) rhesus macaque drinkers. Using an FDR < 0.05, zero genes were significantly differentially expressed (DE) between LD* and controls, six genes between HD* and LD*, and 734 genes between HD* and controls. Focusing on HD* versus control DE genes, the upregulated genes (N = 366) were enriched in genes with annotations associated with signal recognition particle (SRP)-dependent co-translational protein targeting to membrane (FDR < 3 × 10−59), structural constituent of ribosome (FDR < 3 × 10−47), and ribosomal subunit (FDR < 5 × 10−48). Downregulated genes (N = 363) were enriched in annotations associated with behavior (FDR < 2 × 10−4), membrane organization (FDR < 1 × 10−4), inorganic cation transmembrane transporter activity (FDR < 2 × 10−3), synapse part (FDR < 4 × 10−10), glutamatergic synapse (FDR < 1 × 10−6), and GABAergic synapse (FDR < 6 × 10−4). Ingenuity Pathway Analysis (IPA) revealed that EIF2 signaling and mTOR pathways were significantly upregulated in HD* animals (FDR < 3 × 10−33 and <2 × 10−16, respectively). Overall, the data supported our working hypothesis; excessive consumption would be associated with transcriptional differences in GABA/glutamate-related genes.

Effect of chronic binge-like ethanol consumption on subsequent cocaine reinforcement in rhesus monkeys

BackgroundAlthough most individuals with cocaine use disorder also abuse alcohol, little is known about the behavioral and pharmacological mechanisms that promote co-abuse. For example, it is unclear whether prior experience with alcohol renders individuals more sensitive to cocaine when it is subsequently experienced. MethodsThis study examined the effects of chronic ethanol consumption on subsequent cocaine reinforcement in rhesus monkeys. Six monkeys consumed 2.0 g/kg ethanol in a binge-drinking paradigm and 6 monkeys drank a non-alcoholic solution 5 days per week. After 9 months, each monkey’s sensitivity to acquiring cocaine self-administration was determined. Monkeys performed an operant response resulting in food pellet delivery under a fixed-ratio 30 schedule of reinforcement. Saline, then ascending doses of cocaine, were substituted for food pellets until a cocaine dose was reached at which the number of cocaine injections delivered differed significantly from saline injections delivered. Following acquisition, a complete cocaine dose-effect curve was generated to determine whether ethanol consumption altered the reinforcing potency of cocaine determined by calculating the ED50 of the ascending limb of each subject’s curve. ResultsAlthough individual variability was observed, the cocaine dose which initially served as a reinforcer did not differ between ethanol-drinking and control groups and, within the ethanol-drinking group, was not related to the amount of ethanol consumed. Moreover, the reinforcing potency of cocaine did not differ between groups. ConclusionTaken together, the data suggest that a history of binge-like alcohol consumption does not affect sensitivity to cocaine when it is subsequently first experienced.

Sex differences in cholinergic circuits and behavioral disruptions following chronic ethanol exposure with and without thiamine deficiency.

Few studies have investigated differences in the vulnerabilities of males and females to alcohol use disorder and alcohol-related brain damage (ARBD). According to epidemiological and clinical findings, females appear to be more sensitive to the effects of alcohol and thiamine deficiency and have a worse prognosis in recovery from neurocognitive deficits compared with males. This study aimed to characterize the effects of chronic ethanol (EtOH) toxicity and thiamine deficiency across the sexes using rodent models. Male and female Sprague Dawley rats were assigned to chronic forced EtOH treatment (CET), pyrithiamine-induced thiamine deficiency (PTD), combined CET-PTD, or pair-fed (PF) control treatment conditions. Following treatments, spatial working memory was assessed during a spontaneous alternation task while measuring acetylcholine (ACh) in the prefrontal cortex (PFC) and the hippocampus (HPC). The animals also underwent an operant-based attentional set-shifting task (ASST) for the analysis of behavioral flexibility. Female and male rats did not differ in terms of EtOH consumption; however, the CET and CET-PTD-treated female rats had lower BECs than male rats. Compared with the PF group, the CET, PTD, and CET-PTD groups exhibited spatial working memory impairments with corresponding reductions in ACh efflux in the PFC and HPC. The ASST revealed that CET-PTD-treated males and females displayed impairments marked by increased latency to make decisions. Thalamic shrinkage was prominent only in the CET-PTD and PTD treatment conditions, but no sex-specific effects were observed. Although the CET and CET-PTD-treated females had lower BECs than the males, they demonstrated similar cognitive impairments. These results provide evidence that female rats experience behavioral and neurochemical disruptions at lower levels of alcohol exposure than males and that chronic EtOH and thiamine deficiencies produce a unique behavioral profile.

Effects of Chronic Ethanol Consumption and Ovariectomy on the Spontaneous Alveolar Bone Loss in Rats.

Postmenopausal estrogen deficiency and ethanol (EtOH) abuse are known risk factors for different diseases including bone tissues. However, little is known about the synergic effects of EtOH abuse and estrogen deficiency on alveolar bone loss in women. The present study evaluated the effects of EtOH chronic exposure and ovariectomy on the alveolar bone loss in female rats. For this, 40 female Wistar rats were randomly divided into 4 groups: control, EtOH exposure, ovariectomy (OVX), and OVX plus EtOH exposure. Initially, half of the animals were ovariectomized at 75 days of age. After that, the groups received distilled water or EtOH 6.5 g/kg/day (20% w/v) for 55 days via gavage. Thereafter, animals were sacrificed and the mandibles were collected, dissected, and separated into hemimandibles. Alveolar bone loss was evaluated by measuring the distance between the cementoenamel junction and the alveolar bone crest through a stereomicroscope in 3 different anatomical regions of the tissue. One-way ANOVA and post hoc Tukey were used to compare groups (p < 0.05). The results showed that the ovariectomy and EtOH exposure per se were able to induce alveolar bone loss, and their association did intensify significantly the effect. Therefore, OVX associated with heavy EtOH exposure increase the spontaneous alveolar bone loss in rats.

Flightless-I is a potential biomarker for the early detection of alcoholic liver disease

Alcoholic liver disease (ALD) is closely linked to oxidative stress induction. Antioxidant enzymes balance oxidative stress and function as intermediary signaling regulators. Nucleoredoxin (NXN), an antioxidant enzyme, regulates physiological processes through redox-sensitive interactions. NXN interacts with myeloid differentiation primary response gene-88 (MYD88) and flightless-I (FLII) to regulate toll-like receptor 4 (TLR4)/MYD88 pathway activation, but FLII also regulates key cell processes and is secreted into the bloodstream. However, the effects of chronic ethanol consumption recapitulated by either ethanol alone or in combination with lipopolysaccharides (LPS), as a two-hit ALD model, on FLII/NXN/MYD88 complex and FLII secretion have not been explored yet. In this study, we have demonstrated that ethanol feeding increased FLII protein levels, its nuclear translocation and plasma secretion, and modified its tissue distribution both in vivo and in vitro ALD models. Ethanol increased MYD88/FLII interaction ratio, and decreased NXN/MYD88 interaction ratio but this was partially reverted by two-hit model. While ethanol and two-hit model increased MYD88/TLR4 interaction ratio, two-hit model significantly decreased FLII nuclear translocation and its plasma secretion. Ethanol and LPS provoked similar effects in vitro; however, NXN overexpression partially reverted these alterations, and ethanol alone increased FLII secretion into culture medium. In summary, by analyzing the response of FLII/NXN/MYD88 complex during ALD early progression both in vivo and in vitro, we have discovered that the effects of chronic ethanol consumption disrupt this complex and identified FLII as a candidate non-invasive plasma biomarker for the early detection of ALD.

Effects of chronic ethanol consumption on the expression of GLT-1 and neuroplasticity-related proteins in the nucleus accumbens of alcohol-preferring rats

Chronic ethanol exposure induces impairments in CNS excitatory and inhibitory activity. These impairments are associated with glutamatergic dysfunction, including altered neuroplasticity. This study examined the effects of 6-week ethanol (15% and 30% v/v) consumption, by male alcohol-preferring P rats, on protein expression associated with neuroplasticity and glutamate transporter-1 (GLT-1) function. The latter regulates intra- and extra-synaptic glutamate levels. We focused on the shell and core subregions of the nucleus accumbens (Acb); i.e., shell (AcbSh) and core (AcbCo), for these measures. Chronic ethanol exposure increased the expression of BDNF, Arc and phosphorylated (p)-post-synaptic density protein-95 (p-PSD-95) in the AcbSh of P rats. Moreover, the ratio of phospho-neuronal nitric oxide synthase (p-nNOS) to total nNOS was also increased in the AcbSh. These changes in BDNF, Arc and p-nNOS/nNOS ratio were not observed in the AcbCo. Furthermore, chronic ethanol consumption reduced GLT-1 expression in the AcbSh. Alternatively, treatment with ceftriaxone (CEF), a known GLT-1 upregulator, abolished the effect of chronic ethanol consumption on BDNF expression in the AcbSh. Overall, the present findings confirm that chronic ethanol consumption modulates activity-associated synaptic proteins, including BDNF, Arc and nNOS in a subregion-specific (i.e., in the AcbSh but not AcbCo) manner. Thus, alterations in mesocorticolimbic glutamatergic homeostasis and neuroplasticity are possible functional targets for the treatment of alcohol use disorders.

Chronic Alcohol Consumption Exacerbates the Severity of Psoriasiform Dermatitis in Mice.

A relationship between alcohol consumption and psoriasis has been reported, but it is unclear whether alcohol consumption aggravates psoriasis. Here, we studied the effect of chronic ethanol (EtOH) consumption in the murine model of Aldara-induced psoriasiform dermatitis. C57BL/6 mice received 5% EtOH in their drinking water for 10weeks. Dermatitis was induced from weeks 9 to 10, by applying Aldara to the shaved patch of skin on the back. Inflammation was characterized by histological and transcriptomic analyses. EtOH consumption aggravated Aldara-induced dermatitis. The scales were more severe, epidermal thickening was more pronounced, and cutaneous expression of Th17-related cytokines was exacerbated. Control mice simply receiving EtOH displayed minimal cutaneous inflammation, characterized by epidermal infiltrates of T lymphocytes and the overexpression of IL-17A and the Th17-recruiting chemokine CCL20. In vitro studies showed that low concentrations of EtOH induce the expression of CCL20 by murine epidermal keratinocytes. Alcohol consumption leads to subliminar skin inflammation, which is revealed by the exacerbation of Aldara-induced experimental psoriasiform dermatitis, likely through Th17-type minimal skin inflammation. These results favor the systematic management of alcohol consumption in psoriatic patients.

CHRONIC ALCOHOLISM ASSOCIATED WITH DIABETES INDUCED APOPTOSIS IN THE CORPUS CAVERNOSUM IN RATS

The association between diabetes mellitus and erectile dysfunction has been studied for many years. Erectile dysfunction secondary to angiopathic, neuropathic and myopathic damage is one of the main complications of diabetes in men. Other reproductive complications include ejaculatory dysfunction, hypogonadism, modified semen parameters, and delayed puberty. On the other hand, ethanol has various effects too, on male sexual activity, in small amounts improves erection and increases libido because of its vasodilatory effect; however, large amounts can cause central sedation, decreased libido, impaired the penile tumescence, and transient erectile dysfunction. Chronic alcoholism may cause hypogonadism and polineuropathy that may affect penile nerve function. Therefore, chronic alcoholism induced dysfunction of smooth muscle in the corpus cavernosum in rats. Although, effects of ethanol on the ultrastructure of cavernosal smooth muscle cells, elastic fibers and collagen content; and apoptosis in the erectile tissues of diabetic rats has been showed, no studies have been done to analyse the apoptotic effects of the diabetes associated with ethanol in the corpus cavernosum in rats.ObjectivesTo investigate the effects of chronic ethanol consumption and diabetes on apoptotic process in the corpus cavernosum.Material and MethodsMale Wistar rats were divided into four groups: control, diabetic (alloxan injection), ethanol, and ethanol‐diabetic (12 rats in each group). Fragments of corpus cavernosum were obtained and destined to the immunohistochemistry and real‐time PCR to analyze the protein expression of apoptotic genes CASPASE‐3, and CASPASE‐9.ResultsThe immunoreactivity of CASPASE‐3 and CASPASE‐9 was diffuse and higher in the treated groups, however there was no significant difference between the experimental groups, only when compared with the control group. An increase was observed in the gene expression of CASPASE‐3 and CASPASE‐9 in the diabetic and ethanol‐diabetic groups when compared with to control and ethanol groups.ConclusionsThere was an increase in apoptosis in ethanol and ethanol‐diabetic rats. The association of these factors (ethanol and diabetes), jointly with neuropathy and arteriopathy could lead to development of erectile dysfunction.

Chronic ethanol consumption and HBV induce abnormal lipid metabolism through HBx/SWELL1/arachidonic acid signaling and activate Tregs in HBV-Tg mice.

Rationale: Chronic ethanol consumption as a public health problem worldwide boosts the development of chronic liver diseases in hepatitis B virus (HBV)-infected patients. Arachidonic acid metabolite prostaglandin E2 (PGE2) activates regulatory T cells (Tregs) function. Here, we aim to investigate the underlying mechanism by which chronic ethanol consumption enriches the HBV-induced abnormal lipid metabolism and Tregs.Methods: The si-RNAs were used to weaken the expression of SWELL1 in HepG2, HepG2.2.15 and K180 cancer cell lines, followed by RNA sequencing from HepG2 cells. Arachidonic acid metabolite PGE2 and LTD4 were measured by ELISA assay in vivo and in vitro. Western blot analysis and RT-qPCR were used to examine HBx and SWELL1 and transcriptional factor Sp1 in clinical HCC samples and cell lines. The effect of chronic ethanol consumption on Tregs was tested by flow cytometry in HBV-Tg mice. The splenic Tregs were collected and analyzed by RNA sequencing.Results: The cooperative effect of ethanol and HBV in abnormal lipid metabolism was observed in vivo and in vitro. The depression of SWELL1 (or HBx) resulted in the reduction of lipid content and arachidonic acid metabolite, correlating with suppression of relative gene atlas. Ethanol and SWELL1 elevated the levels of PGE2 or LTD4 in the liver of mice and cell lines. Interestingly, the ethanol modulated abnormal lipid metabolism through activating HBx/Sp1/SWELL1/arachidonic acid signaling. Chronic ethanol consumption remarkably increased the population of PBL Tregs and splenic Tregs in HBV-Tg mice, consistently with the enhanced expression of PD-L1 in vivo and in vitro. Mechanically, RNA-seq data showed that multiple genes were altered in the transcriptomic atlas of Tregs sorting from ethanol-fed mice or HBV-Tg mice.Conclusion: The chronic ethanol intake enriches the HBV-enhanced abnormal lipid metabolism through HBx/SWELL1/arachidonic acid signaling and activates Tregs in mice.

Effects of ethanol and ethanol metabolites on intrinsic function of mesenteric resistance arteries.

Evidence suggests that ethanol-induced hypertension is associated with increased cardiovascular responsiveness to vasopressors in vivo and enhanced reactivity of isolated arteries to vasopressors ex vivo. The underlying mechanisms are not well understood and the contribution of ethanol metabolites to vascular effects induced by ethanol consumption are unclear. Mesenteric resistance arteries were harvested from Sprague-Dawley rats. Pressure myography was utilized to test effects of ethanol, acetaldehyde and phosphatidylethanol on myogenic tone and on vasoconstriction induced by phenylephrine, arginine vasopressin (aVP), endothelin-1 and KCl. Ethanol, acetaldehyde and phosphatidylethanol concentrations were monitored during the experiments. Ethanol concentrations in the vessel bath decreased with a half-life of 25min; acetaldehyde and phosphatidylethanol concentrations remained constant. Pretreatment with ethanol dose-dependently increased the potency of phenylephrine to induce vasoconstriction 4-fold (p<0.01). These effects were comparable when arteries were pre-treated with a single dose of ethanol for 30min and when ethanol concentrations were kept constant during 30min and 60min of pretreatment. While ethanol also dose-dependently increased the potency of aVP to induce vasoconstriction 1.7-fold (p<0.05), it did not affect vasoconstriction induced by endothelin-1 or KCl. Acetaldehyde pre-treatment (30 min) dose-dependently increased the potency of phenylephrine to induce vasoconstriction 2.7-fold (p<0.01) but did not affect other vasoconstrictor responses. Phosphatidylethanol did not affect any vasoconstrictor responses. Ethanol and its metabolites did not affect myogenic tone. These data suggest that ethanol and acetaldehyde selectively sensitize intrinsic constrictor responses upon activation of vascular α1-adrenergic and/or vasopressin receptors at clinically relevant concentrations. Our findings support the concept that enhanced vasoreactivity to vasoactive hormones contributes to the development of hypertension induced by ethanol consumption. Ex vivo exposure of resistance arteries to ethanol and acetaldehyde resembles effects of chronic ethanol consumption on intrinsic vascular function, and thus could serve as test platform to evaluate interventions aimed to mitigate vascular effects associated with ethanol consumption.

The Effect of Chronic Ethanol Consumption on Sexual Motivation and Behavior of Adult Male Wistar Rats in the Copulatory Phase.

BackgroundThe interaction of ethanol consumption and sexual behavior has been evaluated over the past three decades; however, some studies have assessed how ethanol consumption affects the general behavioral aspects of the copulatory cycle patterns in male rats. The aim of this study was to investigate the effect of chronic ethanol consumption on adult male Wistar rats' sexual motivation and behavior alteration in pre-copulatory, copulatory, and executive phases of the copulatory cycle.MethodsMale Wistar rats were randomly allocated to two groups (control and ethanol treated groups). After 42 days of treatment, male rats were given access to adult female rats for 2 hours and their sexual behavior were recorded in a fully dark room using an infrared camera.FindingsChronic ethanol consumption caused a significant increase in anogenital sniffing and mounting, intermission, and ejaculation latencies periods, as well as a significant decrease in the sexual activity index (SAI) and copulatory efficiency (CE) compared to the control group.ConclusionIt is suggested that chronic ethanol consumption suppresses sexual behavior and reduces male rats' tendency toward sexual interaction with female rats as manifested by the enhanced latency periods in the copulatory phases and reduced SAI of ethanol treated animals.

Expression Profiles of eNOS, iNOS, endothelin receptors and microRNAs‐27b, 155 and 199 in the corpus cavernosum of rats submitted to chronic alcoholism and diabetes mellitus.

IntroductionEthanol consumption has been described to increase the levels of plasma endothelin‐1 (ET‐1) suggesting that chronic consumption activates endothelinergic pathway and also to affect nitric oxide synthase (NOS) derived nitric oxide (NO) and NOS expression in different tissues. Recent evidence showed that chronic ethanol consumption increases ET‐1‐induced sustained contraction of trabecular smooth muscles cells of the corpora cavernosal in corpus cavernosum of rat by mechanism that involves increased expression of ETA eee and ETB receptors.ObjectivesTo evaluate the effects of chronic ethanol consumption associated with diabetes affected the endothelin system in the cavernosal smooth muscle (CSM) of rats, as well as the effects of chronic ethanol consumption and diabetes on nitric oxide (NO)‐mediated relaxation of cavernosal smooth muscle (CSM).Materials and MethodsCorpus cavernosum were divided into four groups: control, ethanol, diabetic and ethanol‐diabetic of Male Wistar rats. We analyzed protein and gene expression of endothelial NO synthase (eNOS), inducible NO synthase (iNOS) and endothelin receptors (ETA eeeand ETB) by immunohistochemistry and real time‐PCR. The expression of miRNAs‐27a, 199 and 155 by real time‐PCR in corpus cavernosum. It was also evaluated the effect of ethanol consumption associated with diabetes on the relaxation induced by acetylcholine (ACh; 0.01–1000 micromol/L) and in the contraction induced by Endothelin‐1 (ET‐1 0.1 nmol‐ 1μmol L‐1) in organ chambers for measurement of isometric tension.ResultsThe endothelium‐dependent relaxation induced by ACh was decreased in CSM from ethanol, diabetic, ethanol‐diabetic groups when compared with the control group. Immunohistochemistry for eNOS, iNOS, ETA eeeand ETB showed an increase in cavernosal smooth muscle cells in the ethanol, diabetic and ethanol‐diabetic groups when compared with the control group. Similarly, the mRNA levels for eNOs, ETA eeeand ETB were increased in CSM of groups ethanol, diabetic and ethanol‐diabetic and miRNAs‐27a, 155 and 199 were decreased in cavernosal smooth muscle from the ethanol, diabetic and ethanol‐diabetic groups.ConclusionsOur results showed that chronic ethanol consumption associated with diabetes was an impairment of relaxation of CSM from ethanol, diabetic, and ethanol‐diabetic rats that involved a decrease in the NO pathway by endothelium‐dependent mechanisms accompanied by a change in the corpus cavernosum contractile sensitivity. We can also suggest that chronic ethanol consumption associated with diabetes changed the endothelinergic system in the CSM played a role in the pathogenesis of erectile dysfunction, suggesting that these changes of endothelin receptors were regulated by miRNAs‐155, 199.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Astrocyte-specific transcriptome responses to chronic ethanol consumption.

Astrocytes play critical roles in central nervous system (CNS) homeostasis and are implicated in the pathogenesis of neurological and psychiatric conditions, including drug dependence. Little is known about the effects of chronic ethanol consumption on astrocyte gene expression. To address this gap in knowledge, we performed transcriptome-wide RNA sequencing of astrocytes isolated from the prefrontal cortex (PFC) of mice following chronic ethanol consumption. Differential expression analysis revealed ethanol-induced changes unique to astrocytes that were not identified in total homogenate preparations. Astrocyte-specific gene expression revealed calcium-related signaling and regulation of extracellular matrix genes as responses to chronic ethanol use. These findings emphasize the importance of investigating expression changes in specific cellular populations to define molecular consequences of chronic ethanol consumption in mammalian brain.

Rho/rho-kinase signalling in chronically alcohol-fed mice.

The effects of chronic ethanol consumption on male sexual function are debateable, and its effects on the Rho/Rho-kinase pathway are unknown. Therefore, we investigated smooth muscle reactivity and Rho/Rho-kinase signalling in the corpus cavernosum of ethanol-fed mice. Ethanol was added to drinking water at 5% concentration in the first 2 days with 5% increment every subsequent 2 days, up to a final concentration of 20% for 45 days. Corpora cavernosa were isolated and a cumulative dose-response curve to phenylephrine was obtained. Acetylcholine (10-6 M), electrical field stimulation (EFS, 40 V, 8-16 Hz) and Y-27632 (10-6-10-5 M)-induced relaxations were compared in the control and ethanol groups. Blood ethanol levels in the control and ethanol-treated mice were 1.5 ± 0.3 mg/dL and 37.4 ± 4.1 mg/dL (P < 0.001), respectively. Phenylephrine-induced contractile responses were potentiated in the ethanol group, with pD2 values of 4.92 ± 0.18 and 5.71 ± 0.21 (P < 0.01) in the corpus cavernosum obtained from control and alcohol-fed mice, respectively. The relaxant responses to acetylcholine and EFS, but not Y-27632, were significantly augmented in the corpus cavernosum obtained from ethanol-treated mice. However, expression and activation of Rho-kinase remained unchanged in the alcohol group. Ethanol drinking may increase sensitivity to both vasoconstrictors and vasodilators in the mouse corpus cavernosum without any alteration in Rho/Rho-kinase signalling.