Deleterious Effects Of Ethanol Research Articles

D-ribose-L-cysteine exhibits neuroprotective activity through inhibition of oxido-behavioral dysfunctions and modulated activities of neurotransmitters in the cerebellum of Juvenile mice exposed to ethanol

Alcohol exposure to the cerebellum has been known to trigger cerebellar dysfunctions through several mechanisms. This present study was designed to evaluate the repealing effect of D-ribose-L-cysteine (DRLC) on alcohol-induced cerebellar dysfunctions in juvenile BALB/c mice. The animals were randomly divided into 4 groups (n = 10 per group). Mice were given oral administration of normal saline (control), DRLC (100 mg/kg, p.o), ethanol (0.2 mL of 10% w/v), or DRLC (100 mg/kg, p.o) + ethanol (0.2 mL of 10% w/v). On day 29 of the study (i.e., 24 h after the administration of the last respective doses), neurochemical quantification of the respective levels of serotonin and dopamine, lipid peroxidation, total antioxidant, superoxide dismutase, and glutathione peroxidase in the cerebellar tissues of the mice were analyzed. Compared with the saline-treated group, the studied neurochemical indices were modulated across the various experimental groups. The administration of ethanol significantly modulates the levels of monoamine neurotransmitters (serotonin and dopamine) as well as contents of total antioxidants, activities of superoxide dismutase, and glutathione peroxidase, with a concurrently increased level of lipid peroxidase in the cerebellar tissue of the mice. DRLC significantly reverses these effects in the DRLC + ethanol co-treated group. Combined exposure to DRLC + ethanol counteracts the deleterious effect of ethanol in the cerebellum of juvenile BALB/c mice via monoamine neurotransmitter, lipid peroxidation, total antioxidant status, superoxide dismutase, and glutathione peroxidase action pathways. Therefore, DRLC could be a pharmacologic or therapeutic agent in attenuating the deleterious effects of alcohol on the cerebellum.

Blood brain barrier as an interface for alcohol induced neurotoxicity during development

Ethanol consumption during pregnancy or lactation permanently impairs the development of the central nervous system (CNS), resulting in the spectrum of fetal alcohol disorders (FASD). FASD is a general term that covers a set of deficits in the embryo caused by gestational alcohol exposure, with fetal alcohol syndrome (FAS) considered the most serious. The clinical features of FAS include facial abnormalities, short stature, low body weight, and evidence of structural and/or functional damage to the central nervous system (CNS). The prevalence of FAS carriers worldwide is about 15 for every 10,000 live births (about 119,000 children with APS born per year). Epidemiological data in the US show that the incidence of FAS exceeds other congenital syndromes such as Down syndrome and spina bifida. The deleterious effects of ethanol appear in different brain regions, varying according to the dose and period of neural development when the embryo was exposed, and include: 1) microcephaly; 2) abnormalities in cortical development, with a significant decrease in gyrification; 3) agenesis or hypoplasia of the corpus callosum; and 4) cognitive and behavioral deficits (such as impaired memory and learning, speech difficulties, and hyperactivity). Current evidence indicates that CNS blood vessels are particularly affected by teratogenic ethanol. The CNS vasculature is composed of specialized endothelial cells that establish intimate interactions with astrocytes, pericytes, and microglia, constituting the neurovascular unit of the blood-brain barrier (BBB). Together with the fact that BBB exert protective function, it can prevent the passage of substances and drugs to treat diseases that affect the CNS. Pathological changes in the BBB, such as drug abuse during pregnancy, congenital infections, or ageing processes can drastically alter the molecular structure and vascular stability, disrupting the BBB and aggravating certain neurodegenerative and neurological diseases. In this review, we address the effects of alcohol exposure on the formation of the BBB, specifically describing the cellular and molecular events induced by ethanol in the physiology of endothelial cells and glial cells, as well as their interaction during CNS development.

Ethanol: Toxicity and Dangers in Women of Child-Bearing Age

The World Health Organisation estimates that alcohol abuse by adults accounts for about 5% of global disease burden. Additionally, prenatal alcohol exposure (PAE) causes ‘fetal alcohol spectrum disorder’ (FASD). Depending on severity, FASD is characterised by low birth weight, small head size at birth and growth retardation. There are also facial features of narrow eyes, flat upper lip and midface and impaired fine motor skills, hearing loss, poor hand-eye coordination and cognitive impairment. World-wide, up to 10% of children may be affected by PAE. It is unclear what dose or pattern of drinking results in these damaging effects, but animal models suggest that high, acute doses of ethanol (‘binge drinking’) in early pregnancy can result in the facial changes of FASD, whilst sustained, lower dose intake in later pregnancy produces anxiety and depression-like symptoms and deficits of learning and memory. The mechanisms underlying the deleterious effects of PAE are also unresolved, but evidence exists of long-lasting damage due to oxidative stress, increases in inflammatory mediators and changes to the brain renin-angiotensin system. There is also evidence of epigenetic changes. There is a need to prevent or limit the potential adverse effects of ethanol on the unborn child. It is highly unlikely, however, that all sexually-active women of child-bearing age not using reliable contraception will abstain from alcohol. There is therefore a need to research methods of reducing ethanol toxicity for the unborn child and / or develop therapeutic strategies to reverse the deleterious effects of ethanol on the unborn child.

Ethanol: striking the cardiovascular system by harming the gut microbiota.

Ethanol consumption represents a significant public health problem, and excessive ethanol intake is a risk factor for cardiovascular disease (CVD), one of the leading causes of death and disability worldwide. The mechanisms underlying the effects of ethanol on the cardiovascular system are complex and not fully comprehended. The gut microbiota and their metabolites are indispensable symbionts essential for health and homeostasis and therefore, have emerged as potential contributors to ethanol-induced cardiovascular system dysfunction. By mechanisms that are not completely understood, the gut microbiota modulates the immune system and activates several signaling pathways that stimulate inflammatory responses, which in turn, contribute to the development and progression of CVD. This review summarizes preclinical and clinical evidence on the effects of ethanol in the gut microbiota and discusses the mechanisms by which ethanol-induced gut dysbiosis leads to the activation of the immune system and cardiovascular dysfunction. The cross talk between ethanol consumption and the gut microbiota and its implications are detailed. In summary, an imbalance in the symbiotic relationship between the host and the commensal microbiota in a holobiont, as seen with ethanol consumption, may contribute to CVD. Therefore, manipulating the gut microbiota, by using antibiotics, probiotics, prebiotics, and fecal microbiota transplantation might prove a valuable opportunity to prevent/mitigate the deleterious effects of ethanol and improve cardiovascular health and risk prevention.

Polyunsaturated Fatty Acids (PUFA) Increases the Sensitivity of Saccharomyces cerevisiae to Ethanol by Impairing the Function of the Mitochondrial Unselective Channel (MUC)

Yeast cells uncouple its respiration by opening a mitochondrial unselective channel (MUC) that dissipates the membrane potential and accelerates oxygen consumption in order to avoid a deleterious buildup of ROS. Yeast incorporates polyunsaturated fatty acids (PUFA) into membranes when available in its environment, increasing the sensitivity of membranes to oxidative damage. One factor causing oxidative stress in yeast is ethanol accumulation during fermentation. The MUC is constituted by the voltage-dependent anionic channel (VDAC), the adenine nucleotide translocator and the cyclophilin D, being the VDAC a sensor for Ca2+ levels that promotes MUC closing under high Ca2+ concentrations. Moreover, MUC function is dependent on the phospholipid cardiolipin. In this work, we propose that yeast become more susceptible to ethanol stress when grow in the presence of PUFA by the impairment of MUC function due to enhanced lipid peroxidation. Yeast cells were grown in liquid YPD medium with (+C18:3) or without (-C18:3) 1mM linolenic acid and 10% ethanol. Ethanol decreased yeast growth by 20% and 90% in –C18:3 and +C18:3 cells, respectively. Ethanol also promoted lipid peroxidation only in +C18:3 cells. The addition of amiodarone, which increases cytosolic Ca2+ levels and promotes the closure of MUC, decreased 80% the growth in –C18:3 cells, while the +C18:3 cells showed the same levels of growth inhibition than without amiodarone. In – C18:3 cells respiring in uncoupled state (U), amiodarone decreased respiration by promoting MUC closing and coupling. Conversely, respiration was slightly inhibited by amiodarone in the +C18:3 cells. The role of MUC in the deleterious effects of ethanol in +C18:3 cells was confirmed in yeast mutants depleted in cardiolipin and VDAC, as these cells became insensitive to the effects of amiodarone. These results suggest that increased sensitivity to ethanol elicited by C18:3 may be due to impaired MUC function due to enhanced lipid peroxidation.

Deleterious Effects of Ethanol, Δ(9)-Tetrahydrocannabinol (THC), and Their Combination on the Spatial Memory and Cognitive Flexibility in Adolescent and Adult Male Rats in the Barnes Maze Task.

Research demonstrates that adolescents differ from adults in their response to drugs of abuse. The aim of the present study was to examine the influence of ethanol, Δ9-tetrahydrocannabinol hydrochloride (THC), and a combination of these drugs given during adolescence on spatial memory in adolescent and adult rats. Thus, adolescent rats (postnatal day (PND) 30) were subjected to the following groups: 0.9% NaCl; 1.5 g/kg ethanol; 1.0 mg/kg THC; 1.5 g/kg ethanol + 1.0 mg/kg THC. Rats received drug injection four times at three-day intervals. One day after the last injection, half of the treated animals were tested in the Barnes maze task, whereas the remaining animals were tested on PND 70. Results show that there was a significant age effect on spatial memory in the Barnes maze task after these drug administrations. Adolescent animals demonstrated more potent deficits in the spatial learning and memory (probe trial) and in cognitive flexibility (reversal learning) than did adults. However, in adult rats that received these drugs in adolescence, memory decline was observed only after ethanol and ethanol + THC administration. Thus, our results are important in understanding the deleterious impact of THC and/or ethanol abuse during adolescence on memory function across the lifespan (adolescent versus adult).

Resveratrol alleviates ethanol-induced hormonal and metabolic disturbances in the rat.

Resveratrol is a polyphenol found in different plant species and having numerous health-promoting properties in animals and humans. However, its protective action against deleterious effects of ethanol is poorly elucidated. In the present study, the influence of resveratrol (10 mg/kg/day) on some hormones and metabolic parameters was determined in rats ingesting 10 % ethanol solution for two weeks. Blood levels of insulin, glucagon and adiponectin were affected by ethanol, however, resveratrol partially ameliorated these changes. Moreover, in ethanol drinking rats, liver lipid accumulation was increased, whereas resveratrol was capable of reducing liver lipid content, probably due to decrease in fatty acid synthesis. Resveratrol decreased also blood levels of triglycerides and free fatty acids and reduced gamma-glutamyl transferase activity in animals ingesting ethanol. These results show that resveratrol, already at low dose, alleviates hormonal and metabolic changes induced by ethanol in the rat and may be useful in preventing and treating some consequences of alcohol consumption.

Direct effects of ethanol on neuronal differentiation: An in vitro analysis of viability and morphology.

The deleterious effects of ethanol (EtOH) on the brain have been widely described, but its effects on the neuronal cytoskeleton during differentiation have not yet been firmly established. In this context, our aim was to investigate the direct effect of EtOH on cortical neurons during the period of differentiation. Primary cultures of cortical neurons obtained from 1-day-old rats were exposed to EtOH after 7days of culture, and viability and morphology were analyzed at structural and ultrastructural levels after 24-h EtOH exposure. EtOH caused a significant reduction of 73±7% in the viability of cultured cortical neurons, by preferentially inducing apoptotic cellular death. This effect was accompanied by an increase in caspase 3 and 9 expression. Furthermore, EtOH induced a reduction in total dendrite length and in the number of dendrites per cell. Ultrastructural studies showed that EtOH increased the number of lipidic vacuoles, lysosomes and multilamellar vesicles and induced a dilated endoplasmatic reticulum lumen and a disorganized Golgi apparatus with a ring-shape appearance. Microtubules showed a disorganized distribution. Apposition between pre- and postsynaptic membranes without a defined synaptic cleft and a delay in presynaptic vesicle organization were also observed. Synaptophysin and PSD95 expression, proteins pre- and postsynaptically located, were reduced in EtOH-exposed cultures. Overall, our study shows that EtOH induces neuronal apoptosis and changes in the cytoskeleton and membrane proteins related with the establishment of mature synapses. These direct effects of EtOH on neurons may partially explain its effects on brain development.

The developmental perspective of the chronic alcoholism: Who’s holding the burden of proof?

Chronic alcoholism is a public health issue, and several theoretical frameworks have been proposed to explain its nature. The developmental approach to chronic alcoholism has a double contour, with neurobiological theories counting on several aspects of the deleterious effects exerted from ethanol over neural structures. Psychological and neurobiological theories are not intrinsically contradictory to each other. The importance of early experiences and the potential sensory clues leading to ethanol-self administration are integral parts of the developmental neurobiology of an alcoholic. The developmental theories need to consider any further the accumulative and deleterious effects of ethanol during the nervous system maturation.

Gender differences in alcohol-induced neurotoxicity and brain damage

Considerable evidence has demonstrated that women are more vulnerable than men to the toxic effects of alcohol, although the results as to whether gender differences exist in ethanol-induced brain damage are contradictory. We have reported that ethanol, by activating the neuroimmune system and Toll-like receptors 4 (TLR4), can cause neuroinflammation and brain injury. However, whether there are gender differences in alcohol-induced neuroinflammation and brain injury are currently controversial. Using the brains of TLR4+/+ and TLR4−/− (TLR4-KO) mice, we report that chronic ethanol treatment induces inflammatory mediators (iNOS and COX-2), cytokines (IL-1β, TNF-α), gliosis processes, caspase-3 activation and neuronal loss in the cerebral cortex of both female and male mice. Conversely, the levels of these parameters tend to be higher in female than in male mice. Using an in vivo imaging technique, our results further evidence that ethanol treatment triggers higher GFAP levels and lower MAP-2 levels in female than in male mice, suggesting a greater effect of ethanol-induced astrogliosis and less MAP-2+ neurons in female than in male mice. Our results further confirm the pivotal role of TLR4 in alcohol-induced neuroinflammation and brain damage since the elimination of TLR4 protects the brain of males and females against the deleterious effects of ethanol. In short, the present findings demonstrate that, during the same period of ethanol treatment, females are more vulnerable than males to the neurotoxic/neuroinflammatory effects of ethanol, thus supporting the view that women are more susceptible than men to the medical consequences of alcohol abuse.

Jatropha curcas: From Biodiesel Generation to Medicinal Applications

Jatropha curcas (JC) is a multipurpose perennial plant that belongs to the Euphorbiaceae family and is native to arid and semiarid tropical regions worldwide. It has many attributes and considerable potential for renewable energy, fish and livestock feeding. Despite its rich application as a renewable source and for animal feeding, JC has barely been explored for its medicinal potential. Here we review several patents related to JC that show it has been underused for medicinal purposes. For example, only one invention disclosure to date utilizes JC, combined with three other plants, in a preparation for wound healing. Motivated by support from the Brazilian funding agencies and anecdotal accounts in Brazil of the medicinal value of JC, we performed a series of pilot studies that demonstrate that JC is able to protect skeletal muscle cells in vitro against the deleterious effects of ethanol. We were able to determine that JC's effects are mediated by the up regulation of HSP60, a critical mitochondrial heat shock related protein that is essential for intracellular REDOX regulation. Given the fact that ethanol myopathy accounts for more than 50% of all cases of myopathy worldwide, we hope that our studies will sparkle new interest from the scientific community to explore the medicinal properties of Jatropha curcas, including the development of new patents leading to new drugs and new targets for the treatment of muscle diseases and other human diseases.

Neonatal Androgenization Exacerbates Alcohol-Induced Liver Injury in Adult Rats, an Effect Abrogated by Estrogen

Alcoholic liver disease (ALD) affects millions of people worldwide and is a major cause of morbidity and mortality. However, fewer than 10% of heavy drinkers progress to later stages of injury, suggesting other factors in ALD development, including environmental exposures and genetics. Females display greater susceptibility to the early damaging effects of ethanol. Estrogen (E2) and ethanol metabolizing enzymes (cytochrome P450, CYP450) are implicated in sex differences of ALD. Sex steroid hormones are developmentally regulated by the hypothalamic-pituitary-gonadal (HPG) axis, which controls sex-specific cycling of gonadal steroid production and expression of hepatic enzymes. The aim of this study was to determine if early postnatal inhibition of adult cyclic E2 alters ethanol metabolizing enzyme expression contributing to the development of ALD in adulthood. An androgenized rat model was used to inhibit cyclic E2 production. Control females (Ctrl), androgenized females (Andro) and Andro females with E2 implants were administered either an ethanol or isocalorically-matched control Lieber-DeCarli diet for four weeks and liver injury and CYP450 expression assessed. Androgenization exacerbated the deleterious effects of ethanol demonstrated by increased steatosis, lipid peroxidation, profibrotic gene expression and decreased antioxidant defenses compared to Ctrl. Additionally, CYP2E1 expression was down-regulated in Andro animals on both diets. No change was observed in CYP1A2 protein expression. Further, continuous exogenous administration of E2 to Andro in adulthood attenuated these effects, suggesting that E2 has protective effects in the androgenized animal. Therefore, early postnatal inhibition of cyclic E2 modulates development and progression of ALD in adulthood.

Effects of Ethanol on Conformational Changes of Akt Studied by Chemical Cross-Linking, Mass Spectrometry, and 18O Labeling

Although PI3K/Akt signaling that regulates neuronal survival has been implicated in the deleterious effects of ethanol on the central nervous system, underlying molecular mechanisms have not been fully elucidated. Akt-membrane interaction is a prerequisite step for Akt activation since it induces interdomain conformational changes to an open conformer that allows Akt phosphorylation by upstream kinases. In this study, we investigated the effect of ethanol on Akt activation by quantitatively probing Akt conformation using chemical cross-linking, (18)O labeling and mass spectrometry. We found that ethanol at pharmacologically relevant concentrations (20 or 170 mM) directly interacts with Akt and alters the local pleckstrin homology domain configuration near the PIP(3)-binding site. We also found that ethanol significantly impairs subsequent membrane-induced interdomain conformational changes needed for Akt activation. The observed alteration of Akt conformation caused by ethanol during the activation sequence provides a new molecular basis for the effects of ethanol on Akt signaling. The in vitro conformation-based approach employed in this study should also be useful in probing the molecular mechanisms for the action of ethanol or drugs on other signaling proteins, particularly for those undergoing dramatic conformational change during activation processes such as members of AGC kinase super family.

Ethanol causes endoplasmic reticulum stress and impairment of insulin secretion in pancreatic β-cells

Chronic ethanol consumption increases the risk of type 2 diabetes mellitus, and ethanol has been reported to cause insulin resistance and, inconsistently, to reduce insulin secretion. The mechanism(s) underlying the reduction of insulin secretion by ethanol is not known. We used β-cell lines and isolated murine islets to determine the effect of ethanol on insulin content and secretion at low- and high-glucose concentrations, in the presence of KCl, diazoxide, tolbutamide, and regulators of cyclic AMP and protein kinase C (PKC). We also determined the gene expression of insulin; pancreas duodenum homeobox 1; and endoplasmic reticulum (ER) stress markers, such as Chop, ERp57, glucose-regulated protein 78/binding immunoglobulin protein, and inositol 1,4,5-triphosphate receptors. Ethanol reduced insulin secretion by interfering with muscarinic signaling and PKC activation but not the K-ATP channels. In addition, ethanol reduced insulin content and caused ER stress. The deleterious effects of ethanol on β-cells were prevented by 4-methyl pyrazole, an inhibitor of alcohol dehydrogenase, suggesting that ethanol metabolism is required for these effects.

Deleterious Effects of Ethanol on Hematological Parameters and Fertility in Albino Rats

Objective: Hematological disorders including anaemia are prevalent in alcoholics. The study was an attempt to find out the toxic effects of ethanol on blood values and semen parameters of albino rats. Material & Methods: 15 male albino rats with body weight (bwt) of 190 – 220 gm were used for the 2-phase study. 25% ethanol was administered via oral cannula to a group of 5 male rats each at daily dose of 0.6ml/200gm bwt respec-tively for 3 days during phase I. Phase II was a recovery study involving 5 male rats exposed to dose regimen as in phase I, and sacrificed after 3-day withdrawal of treatment. The control group of 5 male rats was given sterile water ad-libitum. Each animal was weighed before sacrifice to obtain difference in bwt relative to its basal value. Blood samples were collected by cardio-puncture from the rats for hematology and serum testosterone at the end of each phase. Semen parameters were determined and compared with controls. Results: Ethanol caused significant reduction (P< 0.05) in the hematological profile as well as in the serum testoster-one and semen parameters of the animals. Discontinuation of the drug use however showed gradual recovery of the depressed indices of the semen, serum testosterone and blood parameters. Conclusion: The ethanol could induce reversible changes in hematological profiles and semen parameters of rats, and by extension man. Hence, the study supported the use of alcohol with caution especially in infertile men and those prone to anemic tendencies. Key Words: Albino rats; ethanol; semen parameters; hematology DOI: 10.3126/ajms.v1i2.3350Asian Journal of Medical Sciences 1 (2010) 37-40

Hepatoprotective activity of Picrorhiza kurroa Royle Ex. Benth extract against alcohol cytotoxicity in mouse liver slice culture

Kutki or Picrorhiza kurroa is a herbal medicinal plant belonging to Scrophulariaceae family and is found in the Himalayan region in India. This herb has been traditionally used in treating liver disorders. The antioxidant properties of P. kurroa were evaluated in vitro using different radical scavenging assays. Furthermore, liver slice culture system was used to test the antioxidant activity of this extract and ethanol was used as a hepatotoxin to generate oxidative stress. Hepatotoxicity was quantified in terms of release of intracellular marker enzymes lactate dehydrogenase, glutamate oxaloacetate transaminase and glutamate pyruvate transaminase. Oxidative stress induced by ethanol and its modulation in the presence of P. kurroa extract was tested by estimating the levels of antioxidant enzymes like catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase and glutathione-S-transferase, and of antioxidant molecules like uric acid and reduced glutathione that were quantitated along with lipid peroxidation. Our results clearly demonstrate that aqueous extract of P. kurroa with high antioxidant activity, as demonstrated using different radical scavenging assays, was effective in suppressing the deleterious effects of ethanol. Addition of P. kurroa aqueous extract along with ethanol restored the activities of antioxidant enzymes and significantly reduced lipid peroxidation. Key words: Ethanol, hepatoprotective activity, liver slice culture, marker enzymes, oxidative stress, Picrorhiza kurroa

Prohibitin is expressed in pancreatic β‐cells and protects against oxidative and proapoptotic effects of ethanol

Pancreatic beta-cell dysfunction is a prerequisite for the development of type 2 diabetes. Alcoholism is a diabetes risk factor and ethanol increases oxidative stress in beta-cells, whereas the mitochondrial chaperone prohibitin (PHB) has antioxidant effects in several cell types. In the present study we investigated whether PHB is expressed in beta-cells and protects these cells against deleterious effects of ethanol, using INS-1E and RINm5F beta-cell lines. Endogenous PHB was detected by western blot and immunocytochemistry. Reactive oxygen species were determined by 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate fluorescence assay, and mitochondrial activity was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) reduction, uncoupling protein 2 expression and ATP production. Cell death was determined by Hoechst 33342 staining, cleaved caspase-3 levels and flow cytometry. PHB was expressed in beta-cells under normal conditions and colocalized with Hoechst 33342 in the nucleus and with the mitochondrial probe Mitofluor in the perinuclear area. In ethanol-treated cells, MTT reduction and ATP production decreased, whereas reactive oxygen species, uncoupling protein 2 and cleaved caspase-3 levels increased. In addition, flow cytometry analysis showed an increase of apoptotic cells. Ethanol treatment increased PHB expression and induced PHB translocation from the nucleus to the mitochondria. PHB overexpression decreased the apoptotic effects of ethanol, whereas PHB knockdown enhanced these effects. The protective effects of endogenous PHB were recapitulated by incubation of the cells with recombinant human PHB. Thus, PHB is expressed in beta-cells, increases with oxidative stress and protects the cells against deleterious effects of ethanol.

Blunted response to low oxygen of rat respiratory network after perinatal ethanol exposure: involvement of inhibitory control

Acute ethanol depresses respiration, but little is known about chronic ethanol exposure during gestation and breathing, while the deleterious effects of ethanol on CNS development have been clearly described. In a recent study we demonstrated that pre- and postnatal ethanol exposure induced low minute ventilation in juvenile rats. The present study analysed in juvenile rats the respiratory response to hypoxia in vivo by plethysmography and the phrenic (Phr) nerve response to ischaemia in situ. Glycinergic neurotransmission was assessed in situ with strychnine application and [(3)H]strychnine binding experiments performed in the medulla. After chronic ethanol exposure, hyperventilation during hypoxia was blunted in vivo. In situ Phr nerve response to ischaemia was also impaired, while gasping activity occurred earlier and recovery was delayed. Strychnine applications in situ (0.05-0.5 microM) demonstrated a higher sensitivity of expiratory duration in ethanol-exposed animals compared to control animals. Moreover, [(3)H]strychnine binding density was increased after ethanol and was associated with higher affinity. Furthermore, 0.2 microM strychnine in ethanol-exposed animals restored the low basal Phr nerve frequency, but also the Phr nerve response to ischaemia and the time to recovery, while gasping activity appeared even earlier with a higher frequency. Polycythaemia was present after ethanol exposure whereas lung and heart weights were not altered. We conclude that chronic ethanol exposure during rat brain development (i) induced polycythaemia to compensate for low minute ventilation at rest; (ii) impaired the respiratory network adaptive response to low oxygen because of an increase in central glycinergic tonic inhibitions, and (iii) did not affect gasping mechanisms. We suggest that ethanol exposure during early life can be a risk factor for the newborn respiratory adaptive mechanisms to a low oxygen environment.

Beneficial effects of nicotinamide on alcohol‐induced liver injury in senescence‐accelerated mice

The deleterious effects of ethanol in senescence-accelerated prone 8 mice (SAMP8) and the protective role of nicotinamide (NAM) against ethanol-induced liver injury were examined. The mice were orally administered 2 g ethanol/kg BW and 200 mg or 500 mg NAM/kg BW three times/week for 10 weeks. Results showed that ethanol elevated activity of alanine aminotransferase (ALT) significantly. Ethanol also enhanced the formation of malondialdehyde (MDA) and protein carbonyls in the liver, whereas ethanol treatment resulted in significantly lower activity of hepatic glutathione peroxidase (GPx), catalase and superoxide dismutase (SOD). Hematoxylin and eosin staining indicated moderate to severe fatty infiltration but not fibrosis. Administration of high NAM (500 mg/kg BW) led to markedly decreased levels of hepatic MDA, protein carbonyls, fatty infiltration and the activity of ALT, and increased activity of GPx, catalase and SOD in the ethanol-fed group. Thus, using SAMP8 as animal model for ethanol-induced liver injury in the aged mice, this study demonstrates that NAM is effective in protecting such damage.

Vitamin E prevents ethanol-induced inflammatory, hormonal, and cytotoxic changes in reproductive tissues

Ethanol causes decreased function of the hypothalamic-pituitary-gonadal (HPG) axis. Ethanol resulted in inflammatory changes in HPG manifested by increased concentrations of pro-inflammatory cytokines. Since, such cytokines have deleterious effects on functions of HPG, it seemed possible that ethanol's suppressive action could be due, at least in part, to this inflammation. Since oxidative stress can cause inflammation, we have used the antioxidant vitamin E to test, whether reducing inflammation might protect reproductive functions from ethanol. Rats were fed an ethanol diet or pair fed identically without ethanol for a 3-week period. For the last 10 days, animals were given 30 IU/kg or 90 IU/kg or vehicle. Ethanol significantly increased hypothalamic, pituitary and testicular TNF-alpha and IL-6, all changes prevented by the higher dose of vitamin E. Also, ethanol induced changes in LHRH, LH, testosterone, and testicular germ cell apoptosis were similarly prevented by vitamin E. These data strikingly show that vitamin E protects the HPG from deleterious effects of ethanol and suggests that the mechanism of this protection might be both anti-inflammatory and antioxidant.