Single Dose Of Ethanol Research Articles

Protective effects of orange (Citrus sinensis L.) peel aqueous extract and hesperidin on oxidative stress and peptic ulcer induced by alcohol in rat

BackgroundMassive alcohol drinking can lead to gastric ulcer. In the present study we investigated the gastroprotective effect of Citrus sinensis peel aqueous extract (CSPE) and Hesperidin (H) in ethanol (EtOH) induced oxidative stress and peptic ulcer in rats.MethodsSeventy adult male Wistar rats were divided into seven groups of 10 each: control, EtOH (4 g/kg b.w.), EtOH + various doses of CSPE (100, 200 and 400 mg/kg, b.w.), EtOH + Hesperidin (50 mg/kg, p.o.) and EtOH + Omeprazole (OM, 20 mg/kg, p.o.). Animals were perorally (p.o.) pre-treated with CSPE during 15 days and intoxicated with a single oral administration of EtOH (4 g/kg b.w.) during 2 h. Gastric ulcer was induced in rats with a single dose of ethanol (EtOH). Ulcer index, gene expression of gastric cyclooxygenase-2 (COX-2), tumor necrosis factor alpha (TNF-α), malondialdhyde (MDA), hydrogen peroxide H2O2 and Thiol groups (−SH) content in stomach and antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and gluthation peroxidise (GPx) were measured. Furthermore, histopathological examinations were performed.ResultsThe results showed that ethanol induced gastric damage, improving oxidative stress markers level such as MDA (121 ± 4.45 nmol/mg proteins) and H2O2 (24.62 ± 1.04 μmol/mg proteins), increased pro-inflammatory cytokine (TNF-α level), as well as the expression of COX-2 in the ethanol group. However, a significant depletion of enzymatic and non-enzymatic antioxidants were observed, such as, GPx (72%), SOD (57.5%), CAT (41.6%) and -SH (50%). The lesions were associated with severe histopathological damage. The both Citrus sinensis peel aqueous extract (CSPE) and hesperidin significantly protect against all gastric damages caused by ethanol administration in rats.ConclusionsWe propose that CSPE and hesperidin exhibit protective effects in EtOH-induced peptic ulcer in rat. This protection might be related in to part its antioxidant properties as well as its opposite effects on some studied intracellular mediators.

C-C chemokine receptor type 5 deficiency exacerbates alcoholic fatty liver disease through pro-inflammatory cytokines and chemokines-induced hepatic inflammation.

Chemokines and chemokine receptors implicated with alcoholic liver disease. Studies have shown that inflammation and oxidative stress induce fat molecules aggregation in liver. We evaluated the relationship between alcoholic fatty liver disease and C-C chemokine receptor 5 (CCR5) and impact of inflammation and oxidative stress in fat molecule deposition. Lieber-DeCarli diet containing ethanol or isocaloric control diets were fed to wild-type and CCR5 knockout mice for 10days and gavaged with a single dose of ethanol or isocaloric maltose dextrin at 11th day. Cytokine, chemokine, and reactive oxygen species levels were measured in liver tissues to study the role of CCR5 in alcoholic fatty liver disease. C-C chemokine receptor type 5 knockout mice exacerbated ethanol-induced liver injury. Serum levels of aspartate aminotransferase and alanine aminotransferase were higher in CCR5 knockout mice than wild-type mice, and CCR5 knockout mice showed more severe lipid accumulation in liver tissue than wild-type mice after ethanol feeding. Increased expressions of pro-inflammatory cytokines TNF-α and IL-6 and chemokines CCL2, CCL3, CCL4, and CCL5 result in exacerbation of hepatitis in CCR5 knockout mice after ethanol feeding. Oxidative stress induced by reactive oxygen species was more severe in CCR5 knockout mice, and increasing level of fatty acid import and decreasing level of lipid degradation resulted in lipid accumulation in ethanol-fed CCR5 knockout mice. Deficiency of CCR5 exacerbates alcoholic fatty liver disease by hepatic inflammation induced by pro-inflammatory cytokines and chemokines and oxidative stress.

A hot water extract of turmeric (Curcuma longa) suppresses acute ethanol-induced liver injury in mice by inhibiting hepatic oxidative stress and inflammatory cytokine production.

Turmeric (Curcuma longa) is a widely used spice that has various biological effects, and aqueous extracts of turmeric exhibit potent antioxidant activity and anti-inflammatory activity. Bisacurone, a component of turmeric extract, is known to have similar effects. Oxidative stress and inflammatory cytokines play an important role in ethanol-induced liver injury. This study was performed to evaluate the influence of a hot water extract of C. longa (WEC) or bisacurone on acute ethanol-induced liver injury. C57BL/6 mice were orally administered WEC (20mg/kg body weight; BW) or bisacurone (60µg/kg BW) at 30min before a single dose of ethanol was given by oral administration (3·0g/kg BW). Plasma levels of aspartate aminotransferase and alanine aminotransferase were markedly increased in ethanol-treated mice, while the increase of these enzymes was significantly suppressed by prior administration of WEC. The increase of alanine aminotransferase was also significantly suppressed by pretreatment with bisacurone. Compared with control mice, animals given WEC had higher hepatic tissue levels of superoxide dismutase and glutathione, as well as lower hepatic tissue levels of thiobarbituric acid-reactive substances, TNF-α protein and IL-6 mRNA. These results suggest that oral administration of WEC may have a protective effect against ethanol-induced liver injury by suppressing hepatic oxidation and inflammation, at least partly through the effects of bisacurone.

Upregulation of SIRT1-AMPK by thymoquinone in hepatic stellate cells ameliorates liver injury

Thymoquinone (TQ) is a biologically active compound isolated from the seeds of Nigella sativa L. (Ranuculaceae). This study investigated the hepato-protective effect of TQ on liver injury through AMP-activated protein kinase (AMPK) signaling in hepatic stellate cells (HSCs). In vitro, TGF-β time-dependently attenuated liver kinase B-1 (LKB1) and AMPK phosphorylation, which were blocked by pretreatment with TQ and AICAR (an activator of AMPK). TQ significantly inhibited collagen-Ι, α-SMA, TIMP-1 and enhanced MMP-13 expression, contributing to prevent TGF-β-induced human HSCs activation. Moreover, TQ induced peroxisome proliferator activated receptor-γ (PPAR-γ) expression, which was inhibited by genetic deletion of AMPK. In vivo, C57BL/6 mice were fed with ethanol diet for 10 days, then administering a single dose of ethanol (5g/kg body weight) via gavage. TQ (20 or 40mg/kg) were given by gavage every day. TQ attenuated the increases in serum aminotransferase and hepatic triglyceride in mice fed with ethanol, while significantly activated LKB1 and AMPK phosphorylation. In addition, TQ enhanced the sirtuin 1 (SIRT1) expression. In conclusion, we demonstrate that AMPK pathway is a key therapeutic target for controlling liver injury and TQ confers hepato-protection against TGF-β-induced the activation of HSCs and ethanol-induced liver injury.

Protective effect of treatment with thiamine or benfotiamine on liver oxidative damage in rat model of acute ethanol intoxication

AimsThe aim of this study was to evaluate possible beneficial effects of treatment with thiamine or benfotiamine in an animal model of acute ethanol intoxication. Main methodsThirty male Wistar rats were separated at random into three groups of 10 animals each: Ethanol (E), Ethanol treated with thiamine (T) and Ethanol treated with benfotiamine (BE). Rats were gavaged with single dose of ethanol (5g/kg, 40% v:v). After 30min of ethanol gavage the animals were treated with thiamine or benfotiamine. Six hours after first gavage, the animals were euthanized and blood and liver samples were collected for ethanol and oxidative stress biomarkers quantification. Key findingsSerum ethanol levels were higher in animals treated with thiamine or benfotiamine while hepatic alcohol levels were higher in animals of the group treated with benfotiamine comparing to controls or thiamine treated groups. The lipid peroxidation biomarkers were diminished for the groups treated with thiamine or benfotiamine comparing to E animals. Concerning protein oxidative damage parameters, they were enhanced for animals treated with benfotiamine in relation to other groups. SignificanceIn conclusion, the treatment with thiamine or benfotiamine even 30min after the massive dose of ethanol has proven to be beneficial against liver damage. Improved results were obtained with benfotiamine in relation to oxidative damage from aqueous compartments.

EP 56. Cerebellar learning and its modifiability by alcohol in myoclonus-dystonia

Objectives To investigate cerebellar learning and its modifiability by alcohol in SGCE mutation-positive myoclonus-dystonia and healthy controls using eyeblink conditioning. Background Myoclonus-dystonia is an inherited neurological movement disorder characterized predominantly by generalized myoclonic jerks and focal or segmental dystonia with a remarkable decrease of symptoms due to alcohol. This effect has raised the question whether the cerebellum could play an important role in the generation of motor symptoms. Methods Delay eyeblink conditioning was assessed in 12 SGCE mutation-positive myoclonus-dystonia patients and 16 control subjects on day 1 before (OFF_C2 state) and on day 2 after a single oral ethanol dose (ON_C2 state), calculated according to the Widmark formula to reach a breath alcohol content of 0.8 ‰ (±0.1 ‰ ). For the conditioning paradigm, an air puff applied to the outer canthus of the eye was preceded by a 540 ms tone, the conditioned stimulus, in 10 learning blocks each including 10 paired stimuli (LT1-LT10). The learning paradigm was followed by three extinction blocks (10 stimuli each), in which the single tone was given only (ET1-ET3). To test for differential effects, a multifactorial repeated measures ANOVA was used with the factors STATE (OFF_C2 and ON_C2), BLOCK (LT1-10 and ET1-3) and the in-between subject factor GROUP (patient and controls). Results The ANOVA showed a significant main effect of the factor GROUP (F1;26) = 7.162, p = 0.013) and BLOCK (F12;312) = 20.181, p p = 0.01) ( Fig. 1 ) indicating that ethanol has a differential effect in myoclonus dystonia compared to controls. Without the influence of alcohol, compared to healthy controls, myoclonus-dystonia patients revealed a significantly lower learning effect (GROUP (F1;26) = 13.255, p = 0.001) and BLOCK (F9;234) = 12.23, p p = 0.013) ( Fig. 1 A LT1-10) and an altered extinction rate (LT 10 to ET 1, patients 41%–22% and controls 79%–32%). Although conditioning rates under the influence of alcohol did not significantly differ when comparing patients and control subjects, both groups showed a reverse effect comparing the learning effect before and after alcohol administration. While the control group had lower conditioning rates under alcohol, myoclonus-dystonia patients showed an increased eye blink conditioning rate under alcohol influence with a main effect of the factor STATE when assessing cerebellar learning before and after alcohol consumption (F1;11) = 7.921, p = 0.017). Conclusion Cerebellar learning is impaired in patients with SGCE mutation-positive myoclonus-dystonia and alcohol administration is able to this improves learning effects. This goes along with the clinically evident improvement of motors symptoms after alcohol intake. This study supports the idea of a crucial role of the cerebellum in generating motor symptoms in myoclonus dystonia.

Preventive effect of α-linolenic acid-rich flaxseed oil against ethanol-induced liver injury is associated with ameliorating gut-derived endotoxin-mediated inflammation in mice

The effects of α-linolenic acid (ALA)-rich flaxseed oil (FDO) against ethanol-induced liver injury and the probable molecular mechanisms in a mouse model of chronic-plus-single-binge ethanol feeding were evaluated. Mice were fed Lieber-DeCarli ethanol or control liquid diets with corn oil (CNO) or flaxseed oil for 10 days. On day 11, mice are gavaged with a single dose of ethanol or maltose dextrin. Ethanol exposure with CNO caused severe liver injury, inflammation and oxidative stress in liver, which were remarkably ameliorated by FDO. FDO supplementation decreased the elevation of plasma endotoxin level, which might be attributed to ameliorating ethanol-induced intestinal barrier dysfunction via upregulating the expressions of tight junction proteins. Additionally, FDO supplementation suppressed endotoxin-triggered inflammation via blocking TLR4/MyD88/NF-κB cascades in liver. These findings suggest that ALA-rich flaxseed oil may have potential to be developed as an effective agent for ethanol-induced liver injury.

Diallyl trisulfide attenuates ethanol-induced hepatic steatosis by inhibiting oxidative stress and apoptosis.

Inhibiting the major characteristics of alcoholic fatty liver (AFL) such as lipid accumulation, oxidative stress and apoptosis is a promising strategy of treating AFL. Diallyl trisulfide (DATS) is the major constituent isolated from garlic, which shows promise in the treatment of chronic liver disease. However, the effects of DATS on ethanol-induced liver injury and the related mechanisms remain unclear. The aim of this study was to evaluate the potential protective effects of DATS on AFL and the potential mechanisms. A single intragastric dose of ethanol was given to rats in vivo, while ethanol-stimulated LO2 cells were used as an in vitro model. Our results demonstrated that DATS prevented ethanol-induced injury, as indicated by the reduced activities of aspartate transaminase (AST) and alanine aminotransferase (ALT) in the serum and culture medium, and inhibition of cell apoptosis. Furthermore, DATS reduced hepatic steatosis by up-regulating the expression of peroxisome proliferator-activated receptor-alpha (PPAR-α) and down-regulating the expression of sterolregulatory element binding protein 1c(SREBP-1c). In addition, DATS alleviated ethanol-induced oxidative stress by enhancing non-enzymatic antioxidant and enzymatic antioxidants contents and by reducing the levels of reactive oxygen species (ROS) and malondialdehyde (MDA). These data collectively revealed that DATS protected ethanol-induced liver injury by inhibiting lipid accumulation and oxidative stress.

Betulin alleviated ethanol-induced alcoholic liver injury via SIRT1/AMPK signaling pathway

The present study was conducted to investigate the protective effect of betulin, a triterpene from the bark of Betula platyphylla Suk, against ethanol-induced alcoholic liver injury and its possible underlying mechanisms. In vitro, human hepatic stellate cell line, LX-2 cells were treated with betulin (6.25, 12.5 and 25μM) prior to ethanol (50mM) for 24h. Cell viability was analyzed by methyl thiazolyl tetrazolium assay, protein expressions were assessed by Western blot. In vivo, we induced alcoholic liver injury in male C57BL/6 mice, placing them on Lieber-DeCarli ethanol-containing diets for 10 days and then administering a single dose of ethanol (5g/kg body weight) via gavage. Betulin (20 and 50mg/kg) were given by gavage every day. In vitro results showed that betulin effectively decreased LX-2 cell viability, attenuated collagen-I, α-smooth muscle actin (α-SMA) levels, activated liver kinase B-1 (LKB1) and adenosine monophosphate-activated protein kinase (AMPK) phosphorylation. Betulin suppressed the expression of sterol regulatory element-binding protein-1 (SREBP-1), and genetic deletion of AMPK blocked the effect of betulin on SREBP-1 in ethanol treated LX-2 cells. In vivo, betulin attenuated the increases in serum aminotransferase and triglyceride levels in the mice fed with chronic-binge ethanol, while significantly inhibited SREBP-1 expression and activated LKB1-AMPK phosphorylation. Additionally, betulin enhanced the sirtuin 1 (SIRT1) expression mediated by ethanol. Taken together, betulin alleviates alcoholic liver injury possibly through blocking the regulation of SREBP-1 on fatty acid synthesis and activating SIRT1-LKB1-AMPK signaling pathway.

Hepato- and neuro-protective effects of watermelon juice on acute ethanol-induced oxidative stress in rats.

Chronic and acute alcohol exposure has been extensively reported to cause oxidative stress in hepatic and extra-hepatic tissues. Watermelon (Citrullus lanatus) is known to possess various beneficial properties including; antioxidant, anti-inflammatory, analgesic, anti-diabetic, anti-ulcerogenic effects. However, there is a lack of pertinent information on its importance in acute alcohol-induced hepato- and neuro-toxicity. The present study evaluated the potential protective effects of watermelon juice on ethanol-induced oxidative stress in the liver and brain of male Wistar rats. Rats were pre-treated with the watermelon juice at a dose of 4 ml/kg body weight for a period of fifteen days prior to a single dose of ethanol (50%; 12 ml/kg body weight). Ethanol treatment reduced body weight gain and significantly altered antioxidant status in the liver and brain. This is evidenced by the significant elevation of malondialdehyde (MDA) concentration; depletion in reduced glutathione (GSH) levels and an increased catalase (CAT) activity in the brain and liver. There was no significant difference in the activity of glutathione peroxidase (GPX) in the liver and brain.Oral administration of watermelon juice for fifteen (15) days prior to ethanol intoxication, significantly reduced the concentration of MDA in the liver and brain of rats. In addition, water melon pre-treatment increased the concentration of GSH and normalized catalase activity in both tissues in comparison to the ethanol control group. Phytochemical analysis revealed the presence of phenol, alkaloids, saponins, tannins and steroids in watermelon juice. Our findings indicate that watermelon juice demonstrate anti-oxidative effects in ethanol-induced oxidation in the liver and brain of rats; which could be associated with the plethora of antioxidant phyto-constituents present there-in.

EFFECTS OF LUDARTIN AND LEUKOMISIN ON THE ACUTE HYPERLIPIDEMIA MODEL INDUCED BY ETHANOL

Objective: study sesquiterpene lactones ludartin and leukomisin lipid-lowering properties on the model of acute hyperlipidemia induced by ethanol in rats. Material and methods. Rats during 7 days injected into the stomach ludartin and leukomisin in a dose 10 mg/kg or reference drug nicotinic acid in a dose 25 mg/kg. Hyperlipidemia caused by single introduc-tion of ethanol into the stomach in a dose 5 g/kg. In blood serum of tail vein measured the triacylgly-cerols, total cholesterol, high density and low density lipoproteins cholesterol, also the level of free fatty acids. Calculated the ratio of high density lipoproteins cholesterol to the amount of low density lipopro-teins cholesterol and the index of atherogenicity. Results. A single dose of ethanol increased serum level of triacylglycerols in 1.9 times, free fatty acids – in 3.2 times, low density lipoproteins – on 44% in comparison with the intact animals indices. It shows the development of acute hyperlipidemia. Serum total cholesterol, high density lipoproteins cholesterol and the index of atherogenicity were not changed. Course introduction of sesquiterpene lactones ludartin and leukomisin against the background of acute hyperlipidemia was accompanied by a decrease in the serum of triacylglycerols levels respectively by 37.5% and 49.5%. Nicotinic acid lowered the content of triacylglycerols by 42.4%. Ludartin, leukomisin and nicotinic acid reduced the increased level of free fatty acids in the blood serum by 63.4%, 41.6% and 67.9%. Ludartin, leukomisin and nicotinic acid de-creased by 15.8%, 20.3% and 17.2% of total cholesterol in the blood serum. In acute hyperlipidemia ludartin and leukomisin reduced low density lipoproteins cholesterol by 23.8% and 14.8%, respectively, nicotinic acid – by 15.7%. Both of sesquiterpene lactone and nicotinic acid did not modify the content of high density lipoproteins cholesterol. When introduction ludartin and nicotinic acid ratio of high density lipoproteins cholesterol to the amount of low density lipoproteins cholesterol significantly increased by 32.7% and 22.1% respectively. Conclusion. Sesquiterpene lactones ludartin and leukomisin posses hypolipidemic effect in acute experi-mental hyperlipidemia caused by the ethanol introduction. Lactones normalize many indices of lipid me-tabolism, which can be caused by different biochemical targets of these molecules. Lactones, as nicotinic acid, in the model of acute hyperlipidemia decrease in blood serum triacylglycerols, total cholesterol, and low density lipoproteins cholesterol content.

Metabolic danger signals, uric acid and ATP, mediate inflammatory cross-talk between hepatocytes and immune cells in alcoholic liver disease.

Inflammation defines the progression of ALD from reversible to advanced stages. Translocation of bacterial LPS to the liver from the gut is necessary for alcohol-induced liver inflammation. However, it is not known whether endogenous, metabolic danger signals are required for inflammation in ALD. Uric acid and ATP, 2 major proinflammatory danger signals, were evaluated in the serum of human volunteers exposed to a single dose of ethanol or in supernatants of primary human hepatocytes exposed to ethanol. In vitro studies were used to evaluate the role of uric acid and ATP in inflammatory cross-talk between hepatocytes and immune cells. The significance of signaling downstream of uric acid and ATP in the liver was evaluated in NLRP3-deficient mice fed a Lieber-DeCarli ethanol diet. Exposure of healthy human volunteers to a single dose of ethanol resulted in increased serum levels of uric acid and ATP. In vitro, we identified hepatocytes as a significant source of these endogenous inflammatory signals. Uric acid and ATP mediated a paracrine inflammatory cross-talk between damaged hepatocytes and immune cells and significantly increased the expression of LPS-inducible cytokines, IL-1β and TNF-α, by immune cells. Deficiency of NLRP3, a ligand-sensing component of the inflammasome recognizing uric acid and ATP, prevented the development of alcohol-induced liver inflammation in mice and significantly ameliorated liver damage and steatosis. Endogenous metabolic danger signals, uric acid, and ATP are involved in inflammatory cross-talk between hepatocytes and immune cells and play a crucial role in alcohol-induced liver inflammation.

Binge ethanol prior to traumatic brain injury worsens sensorimotor functional recovery in rats.

A significant number of patients suffering from traumatic brain injury (TBI) have a high blood alcohol level at the time of injury. Furthermore, drinking alcohol in a binge-like pattern is now recognized as a national problem, leading to a greater likelihood of being injured. Our objective was to determine the consequences of a binge paradigm of alcohol intoxication at the time of TBI on long-term functional outcome using a sensitive test of sensorimotor function. We trained adult, male, Sprague Dawley rats on the skilled forelimb reaching task and then administered a single binge dose of ethanol (2g/kg, i.p.) or saline for three consecutive days (for a total of 3 doses). One hour after the final ethanol dose, rats underwent a TBI to the sensorimotor cortex corresponding to the preferred reaching forelimb. Animals were then tested for seven weeks on the skilled forelimb reaching task to assess the profile of recovery. We found that the group given ethanol prior to TBI displayed a slower recovery curve with a lower recovery plateau as compared to the control group. Therefore, even a relatively short (3 day) episode of binge alcohol exposure can negatively impact long-term recovery from a TBI, underscoring this significant public health problem.

Effects of a Single Dose of Ethanol on Survival Rate and Angiogenesis of Chick Embryo

The developing Chick has been widely used as a model organism for research studies in developmental biology. Various aspects of ethanol (EtOH) toxicity during embryonic development have been documented in experimental studies. The studies on chick embryo address the effects of EtOH on craniofacial abnormalities and angiogenesis. The purpose of this study was to assess the effects of a single dose of ethanol on survival rate and angiogenesis of chick embryos during early stages of development. Chick eggs were incubated at 37oC in the humidified incubator, after 72 hours of incubation these eggs were removed from incubator, and treated with a single doses of 1%, 5%, 10%, 13% and 15% EtOH. The effects on survival rate and angiogenesis were recorded on ethanol treated chick embryos. The survival rate was slightly reduced with 1% and 5% EtOH, however with increasing dose of 10% the survival rate was reduced to 64% and at 15% EtOH complete death was observed. Our results also indicate that treatment of EtOH with 1% and 5% did not have any obvious effects on vessels formation in comparison with the BSS treated embryos. However, treatment of chick embryo with 10%, 13% and 15% EtOH severely inhibited the blood vessels formation.

Ethanol impairs intestinal barrier function in humans through mitogen activated protein kinase signaling: a combined in vivo and in vitro approach.

BackgroundEthanol-induced gut barrier disruption is associated with several gastrointestinal and liver disorders.AimSince human data on effects of moderate ethanol consumption on intestinal barrier integrity and involved mechanisms are limited, the objectives of this study were to investigate effects of a single moderate ethanol dose on small and large intestinal permeability and to explore the role of mitogen activated protein kinase (MAPK) pathway as a primary signaling mechanism.MethodsIntestinal permeability was assessed in 12 healthy volunteers after intraduodenal administration of either placebo or 20 g ethanol in a randomised cross-over trial. Localization of the tight junction (TJ) and gene expression, phosphorylation of the MAPK isoforms p38, ERK and JNK as indicative of activation were analyzed in duodenal biopsies. The role of MAPK was further examined in vitro using Caco-2 monolayers.ResultsEthanol increased small and large intestinal permeability, paralleled by redistribution of ZO-1 and occludin, down-regulation of ZO-1 and up-regulation of myosin light chain kinase (MLCK) mRNA expression, and increased MAPK isoforms phosphorylation. In Caco-2 monolayers, ethanol increased permeability, induced redistribution of the junctional proteins and F-actin, and MAPK and MLCK activation, as indicated by phosphorylation of MAPK isoforms and myosin light chain (MLC), respectively, which could be reversed by pretreatment with either MAPK inhibitors or the anti-oxidant L-cysteine.ConclusionsAdministration of moderate ethanol dosage can increase both small and colon permeability. Furthermore, the data indicate a pivotal role for MAPK and its crosstalk with MLCK in ethanol-induced intestinal barrier disruption.Trial RegistrationClinicalTrials.gov NCT00928733

Inhibition of heat shock protein 90 alleviates steatosis and macrophage activation in murine alcoholic liver injury

Heat shock protein 90 (hsp90) is an emerging therapeutic target in chronic liver diseases. Hsp90 plays an important role in liver immune cell activation; however its role in alcoholic liver disease (ALD) remains elusive. Here we hypothesize that hsp90 is crucial in alcohol induced steatosis and pro-inflammatory cytokine production. To test this hypothesis, we employed a pharmacological inhibitor of hsp90, 17-DMAG (17-Dimethylamino-ethylamino-17-demethoxygeldanamycin) in an in vivo mouse model of acute and chronic alcoholic liver injury. C57BL/6 mice were given either a single dose of ethanol via oral gavage (acute) or chronically fed alcohol for 2 weeks followed by oral gavage (chronic-binge). 17-DMAG was administered during or at the end of feeding. Liver injury parameters, inflammatory cytokines and lipid metabolism genes were analysed. Our results reveal increased expression of hsp90 in human and mouse alcoholic livers. In vivo inhibition of hsp90, using 17-DMAG, not only prevented but also alleviated alcoholic liver injury, determined by lower serum ALT, AST and reduced hepatic triglycerides. Mechanistic analysis showed that 17-DMAG decreased alcohol mediated oxidative stress, reduced serum endotoxin, decreased inflammatory cells, and diminished sensitization of liver macrophages to LPS, resulting in downregulation of CD14, NFκB inhibition, and decreased pro-inflammatory cytokine production. Hsp90 inhibition decreased fatty acid synthesis genes via reduced nuclear SREBP-1 and favoured fatty acid oxidation genes via PPARα. Inhibition of hsp90 decreased alcohol induced steatosis and pro-inflammatory cytokines and inhibited alcoholic liver injury. Hsp90 is therefore relevant in human alcoholic cirrhosis and a promising therapeutic target in ALD.

Plasticity of GABAAreceptor-mediated neurotransmission in the nucleus accumbens of alcohol-dependent rats

Chronic alcohol exposure-induced changes in reinforcement mechanisms and motivational state are thought to contribute to the development of cravings and relapse during protracted withdrawal. The nucleus accumbens (NAcc) is a key structure of the mesolimbic dopaminergic reward system and plays an important role in mediating alcohol-seeking behaviors. Here we describe the long-lasting alterations of γ-aminobutyric acid type A receptors (GABA(A)Rs) of medium spiny neurons (MSNs) in the NAcc after chronic intermittent ethanol (CIE) treatment, a rat model of alcohol dependence. CIE treatment and withdrawal (>40 days) produced decreases in the ethanol and Ro15-4513 potentiation of extrasynaptic GABA(A)Rs, which mediate the picrotoxin-sensitive tonic current (I(tonic)), while potentiation of synaptic receptors, which give rise to miniature inhibitory postsynaptic currents (mIPSCs), was increased. Diazepam sensitivity of both I(tonic) and mIPSCs was decreased by CIE treatment. The average magnitude of I(tonic) was unchanged, but mIPSC amplitude and frequency decreased and mIPSC rise time increased after CIE treatment. Rise-time histograms revealed decreased frequency of fast-rising mIPSCs after CIE treatment, consistent with possible decreases in somatic GABAergic synapses in MSNs from CIE rats. However, unbiased stereological analysis of NeuN-stained NAcc neurons did not detect any decreases in NAcc volume, neuronal numbers, or neuronal cell body volume. Western blot analysis of surface subunit levels revealed selective decreases in α1 and δ and increases in α4, α5, and γ2 GABA(A)R subunits after CIE treatment and withdrawal. Similar, but reversible, alterations occurred after a single ethanol dose (5 g/kg). These data reveal CIE-induced long-lasting neuroadaptations in the NAcc GABAergic neurotransmission.

Safety, tolerability, and pharmacokinetic evaluation of single‐ and multiple‐ascending doses of a novel kappa opioid receptor antagonist LY2456302 and drug interaction with ethanol in healthy subjects

Accumulating evidence indicates that selective antagonism of kappa opioid receptors may provide therapeutic benefit in the treatment of major depressive disorder, anxiety disorders, and substance use disorders. LY2456302 is a high-affinity, selective kappa opioid antagonist that demonstrates >30-fold functional selectivity over mu and delta opioid receptors. The safety, tolerability, and pharmacokinetics (PK) of LY2456302 were investigated following single oral doses (2-60 mg), multiple oral doses (2, 10, and 35 mg), and when co-administered with ethanol. Plasma concentrations of LY2456302 were measured by liquid chromatography-tandem mass spectrometry method. Safety analyses were conducted on all enrolled subjects. LY2456302 doses were well-tolerated with no clinically significant findings. No safety concerns were seen on co-administration with ethanol. No evidence for an interaction between LY2456302 and ethanol on cognitive-motor performance was detected. LY2456302 displayed rapid oral absorption and a terminal half-life of approximately 30-40 hours. Plasma exposure of LY2456302 increased proportionally with increasing doses and reached steady state after 6-8 days of once-daily dosing. Steady-state PK of LY2456302 were not affected by coadministration of a single dose of ethanol. No clinically important changes in maximum concentration (Cmax ) or AUC of ethanol (in the presence of LY2456302) were observed.

Evaluation of the safety, pharmacodynamic, and pharmacokinetic effects following oral coadministration of immediate-release morphine with ethanol in healthy male participants.

This paper aimed to evaluate the effects of coadministered immediate-release morphine and ethanol on safety, pharmacokinetic, and pharmacodynamic measures. In the first stage of a randomized, double-blind, placebo-controlled, crossover study, 16 healthy men with a history of moderate drinking received morphine 50 mg+ethanol 0.7 g/kg, morphine 50 mg+ethanol placebo, and morphine placebo+ethanol 0.7 g/kg. In the second stage, participants received either a lower (30 mg) or higher (80 mg) morphine dose (alone and in combination with ethanol) depending on their tolerability to treatments in stage 1. Safety, pharmacodynamic (including visual analog scales, pupillometry, capnography, and psychomotor and cognitive measures), and pharmacokinetic assessments were conducted. With the exception of one severe adverse event (AE), all others were mild or moderate in intensity. Morphine resulted in dose-related increases in AEs. When morphine was administered with ethanol, similar AEs were observed (dizziness, headache, somnolence, nausea, and vomiting), but these were sometimes more frequent compared with those observed with either drug alone. No consistent additive or interaction effects were observed on pharmacodynamic measures. Ethanol had no apparent effects on the pharmacokinetics of morphine or its metabolites. Coadministration of single doses of morphine and ethanol tested in this study did not affect the safety, pharmacodynamics, or pharmacokinetics of morphine or ethanol administered alone. Copyright © 2014 John Wiley & Sons, Ltd.

EFFECTS OF GROSSHEMIN AND GROSSMISIN ON THE ACUTE HYPERLIPIDEMIA MODEL INDUCED BY ETHANOL

Objective: study sesquiterpene lactones grosshemin and grossmisin lipid-lowering properties on the model of acute hyperlipidemia induced by ethanol in rats.Materials and methods. Rats during 7 days injected into the stomach grosshemin and grossmisin in a dose 10 mg/kg or reference drug nicotinic acid in a dose 25 mg/kg. Hyperlipidemia caused by single introduction of ethanol into the stomach in a dose 5 g/kg. In blood serum of tail vein measured the triacylglycerols, total cholesterol, high density and low density lipoproteins cholesterol, also the level of free fatty acids. Calculated the ratio of high density lipoproteins cholesterol to the amount of low density lipoproteins cholesterol and the index of atherogenicity.Results. A single dose of ethanol increased serum level of triacylglycerols in 1.9 times, free fatty acids – in 3.2 times, low density lipoproteins – on 44% in comparison with the intact animals indices. It shows the development of acute hyperlipidemia. Serum total cholesterol, high density lipoproteins cholesterol and the index of atherogenicity were not changed. Course sesquiterpene lactones grosshemin and grossmisin introduction against the background of acute hyperlipidemia was accompanied by a decrease in the serum of triacylglycerols levels respectively by 19.8% and 34.1%. Nicotinic acid lowered the content of triacylglycerols by 42.4%. Grosshemin and nicotinic acid reduced the increased level of free fatty acids in the blood serum by 60.7–67.9%. Grossmisin and nicotinic acid decreased by 14.6–17.2% of total cholesterol in the blood serum. In acute hyperlipidemia grosshemin and grossmisin reduced low density lipoproteins cholesterol by 17.6% and 20%, respectively, nicotinic acid – by 15.7%. Both of sesquiterpene lactone and nicotinic acid did not modify the content of high density lipoproteins cholesterol. When introduction grosshemin, grossmisin and nicotinic acid ratio of high density lipoproteins cholesterol to the amount of low density lipoproteins cholesterol significantly increased by 42.8%, 38,6% and 22.1% respectively.Conclusion. Sesquiterpene lactones grosshemin and grossmisin posses hypolipidemic effect in acute experimental hyperlipidemia caused by the ethanol introduction. Lactones normalize many indices of lipid metabolism, which can be caused by different biochemical targets of these molecules. Lactones, as nicotinic acid, in the model of acute hyperlipidemia decrease in blood serum triacylglycerols, total cholesterol, and low density lipoproteins cholesterol content. Grosshemin and nicotinic acid also reduce the free fatty acids level.