Effect of ethanol and diazepam given to pregnant rats on the behavior and catecholamine content in the brain of offspring

Effect of separate and combined treatment with ethanol and diazepam applied to pregnant rats upon the behavior of 3- and 6-week old and 3 month old rats, as well as on the content of noradrenaline (NA), 5-hydroxytryptamine (5-HT), and 5-hydroxyindoleacetic acid (5-HIAA) in various brain areas was investigated. It has been shown that separate application of ethanol and diazepam causes slight changes in the behavior and content of amines on the brain of offsprings. Combined application of ethanol and diazepam to pregnant rats exerts marked effects upon the behavior of offsprings and causes a decreased content mainly of 5-HT and 5-HIAA in the investigated areas of the brain.     

Effects of ethanol on pregnant rats and their offspring

Pregnant rats were intubated with either 1.0 or 2.0 g/kg of ethanol daily throughout gestation. Pair-fed vehicle-treated, and nontreated rats fed ad libitum, served as control groups for ethanol-treated animals. Ethanol treatment reduced food and water consumption and attenuated the gain in body weight of pregnant animals relative to nontreated animals fed ad libitum. Litter size, litter weight, and the mean weight per pup were reduced in both the ethanol-treated and pair-fed control groups. There was no evidence of gross malformations in any of the offspring. Since the reduction in litter size and litter weights did not differ significantly between ethanol-treated and pair-fed controls, the effects of treatment with ethanol appeared to be related to a reduction in maternal intake of calories rather than to the direct effect of ethanol on the developing fetus.There were no significant differences between any of the groups of offspring on one-way shock avoidance learning, water maze escape learning, spontaneous alternation, or brightness discrimination learning in tests beginning at 75 days of age. Thus, at the doses of alcohol used in this study, there was no evidence of behavioral teratogenesis comparable to that reported for higher doses in animals or in man characterized by the fetal alcohol syndrome.     

Effect of ethanol and toliprolol on the carbohydrate metabolism of the brain and liver

After application of ethanol (1.5 mg/g i.v.) to mice, the content of glycogen of the liver and the brain decreased. The alterated content of the metabolites of the glucose break-down points to an augmentation of the glycolysis in the liver and a diminution of the glycolysis in the brain. By pretreatment with the beta-sympatholytic agent 1-isopropylamino-3-(3-tolyloxy)-2-propanol (toliprolol, Doberol) (50 microgram/g s.c.), the ethanol-induced break-down of glycogen was partially blocked in the liver and completely abolished in the brain. In contrast, mainly synergistic effects of ethanol and toliprolol on the glycolysis of the liver and the brain were found. From the results it is concluded that the ethanol-induced break-down of glycogen is mediated, at least in part, by catecholamines whereas the alteration of the glycolysis by ethanol is brought about by some different mechanism.     

Effect of ethanol on brain neuropeptides in adolescent and adult rats

OBJECTIVE: Alcohol misuse early in life is associated with an increased risk of alcoholism. It is possible that this increased risk in adolescent drinkers is in part related to the susceptibility of the adolescent brain to ethanol. This study assessed the effects of ethanol exposure on several neuropeptides to begin to elucidate potential substrates that could mediate the differential effects of ethanol on adolescent and adult rats. METHOD: Male Sprague Dawley rats were exposed to ethanol vapor or air during adolescence (30 days old, n = 9, controls = 8) or adulthood (80-90 days old, n = 9, controls = 8) for 10 days. Blood alcohol concentrations averaging 250 mg/dl were maintained during this period. After 7 weeks of cessation from ethanol vapor, brain tissue was collected from the frontal cortex, caudate, hippocampus, amygdala and hypothalamus to assess the immunoreactivity levels of neuropeptide Y (NPY-LI), corticotropin-releasing hormone, substance P (SP-LI) and neurokinins (NK-LI). RESULTS: Ethanol exposure decreased overall hippocampal NPY-LI and increased SP-LI and NK-LI in the caudate, but these effects were more prominent in adult rats. Rats in the adult treatment groups (both ethanol exposed and controls) also had significantly lower levels of frontal cortical NK-LI, frontal cortical SP-LI and hypothalamic SP-LI relative to rats in the adolescent treatment groups. CONCLUSIONS: These data indicate that brief exposure to alcohol has long-term effects on levels of NPY-LI, SP-LI and NK-LI. As these effects were primarily the result of changes in rats exposed to ethanol during adulthood, however, they are unlikely to contribute to the increased susceptibility of adolescents to the effects of chronic ethanol exposure.     

泽泻汤对高脂血症大鼠肝脏线粒体蛋白质组的影响

目的 研究泽泻汤对高脂血症大鼠肝脏线粒体蛋白质组的影响.方法 将40只健康雄性SD大鼠随机分为正常组、高脂模型组、辛伐他汀组、泽泻汤组,每组10只,造模4周后辛伐他汀组和泽泻汤组分别灌胃给予相应的药物,正常组和模型组灌胃给予等量的生理盐水.连续给药4周后检测血清TC、TG、LDL-C及HDL-C水平.运用同位素标记的相对与绝对定量技术(isobaric tags for relative and absolute quantitation,iTRAQ)检测大鼠肝脏蛋白质组的变化.结果 高脂模型组大鼠TC、TG、LDL-C水平明显高于正常组(P＜0.05),辛伐他汀组和泽泻汤组给药4周后,TC、TG、LDL-C水平均降低(P＜0.05);iTRAQ分析结果显示,在各组大鼠肝脏组织样本中共检测出4 213种差异蛋白,其中模型组蛋白的变化有统计学意义的共有195种,经泽泻汤治疗后,与肝脏线粒体相关的蛋白表达变化有统计学意义的有18种,其中△3-△2-烯酰-辅酶A异构酶(3,2 trans-enoyl-Coenzyme A isomerase,ECI)表达差异最为显著.结论 泽泻汤对高脂血症大鼠肝脏线粒体能量代谢相关蛋白有影响,泽泻汤降血脂作用可能与线粒体ECI等蛋白差异表达有关.     

Effect of epinephrine and alprenolol on ethanol metabolism, liver cell respiration and mitochondrial function.

Chronic administration of epinephrine to adult male rats resulted in a significant increase in the rate of ethanol elimination, when given alone or together with the beta-adrenergic blocker alprenolol. This effect was observed concomitantly with an increased hepatic oxygen utilization and no changes in mitochondrial respiratory functions. Epinephrine given acutely did not modify the rate of ethanol metabolism. Blood glucose levels were enhanced in these conditions, but were unaffected in rats treated with epinephrine plus alprenolol. These results suggest that chronic epinephrine treatment induces an increased oxidative capacity in the liver characterized by enhanced rates of oxygen uptake and ethanol metabolism, which is not related to its beta-adrenergic actions.     

Prenatal ethanol exposure programs an increased susceptibility of non-alcoholic fatty liver disease in female adult offspring rats

Prenatal ethanol exposure (PEE) induces dyslipidemia and hyperglycemia in fetus and adult offspring. However, whether PEE increases the susceptibility to non-alcoholic fatty liver disease (NAFLD) in offspring and its underlying mechanism remain unknown. This study aimed to demonstrate an increased susceptibility to high-fat diet (HFD)-induced NAFLD and its intrauterine programming mechanisms in female rat offspring with PEE. Rat model of intrauterine growth retardation (IUGR) was established by PEE, the female fetus and adult offspring that fed normal diet (ND) or HFD were sacrificed. The results showed that, in PEE + ND group, serum corticosterone (CORT) slightly decreased and insulin-like growth factor-1 (IGF-1) and glucose increased with partial catch-up growth; In PEE + HFD group, serum CORT decreased, while serum IGF-1, glucose and triglyceride (TG) increased, with notable catch-up growth, higher metabolic status and NAFLD formation. Enhanced liver expression of the IGF-1 pathway, gluconeogenesis, and lipid synthesis as well as reduced expression of lipid output were accompanied in PEE + HFD group. In PEE fetus, serum CORT increased while IGF-1 decreased, with low body weight, hyperglycemia, and hepatocyte ultrastructural changes. Hepatic IGF-1 expression as well as lipid output was down-regulated, while lipid synthesis significantly increased. Based on these findings, we propose a “two-programming” hypothesis for an increased susceptibility to HFD-induced NAFLD in female offspring of PEE. That is, the intrauterine programming of liver glucose and lipid metabolic function is “the first programming”, and postnatal adaptive catch-up growth triggered by intrauterine programming of GC-IGF1 axis acts as “the second programming”.     

Effect of prenatal ethanol exposure during the brain growth spurt of the guinea pig

This study tested the hypothesis that prenatal ethanol exposure during the last third of gestation, including the brain growth spurt (BGS), in the guinea pig produces neurobehavioural teratogenicity, manifesting as brain growth restriction and hyperactivity. Pregnant guinea pigs (term, about gestational day (GD) 68) received oral administration of ethanol (2 g/kg maternal body weight per day on GD 43 and/or GD 44 and then 4 g/kg maternal body weight per day from GD 45 to GD 62), isocaloric-sucrose/pair-feeding, or water. Maternal blood ethanol concentration (BEC) on GD 57 or 58, at 1 h after the daily dose, was 340+/-76 mg/dl (n=8). Ethanol treatment decreased brain, cerebral cortical, hippocampal, and cerebellar weights at GD 63 (P<0.05), and decreased brain and cerebral cortical weights at postnatal day 10 (P<0.05), with no effect on body weight and no apparent effect on spontaneous locomotor activity. The data demonstrate that, in the guinea pig, prenatal ethanol exposure during the last third of gestation, including the BGS, decreases brain weight that persists into postnatal life, which is associated with growth restriction of the cerebral cortex. However, this prenatal ethanol exposure regimen, including the BGS, does not increase spontaneous locomotor activity in contrast to the persistent hyperactivity that occurs after chronic ethanol exposure throughout gestation.     

Effect of green tea extracts on liver functions in Wistar rats.

An herbal medicinal product (Exolise) containing as active ingredient an hydro-alcoholic extract of green tea named AR25 (standardized to 25% catechins) has been implicated in hepatic failures, leading to the withdrawal of the marketing authorization. The active ingredient of Exolise being manufactured with 80% ethanol, the question to know whether the extraction solvent could introduce some toxic components was hypothesized. Two investigations were conducted in Wistar rats to determine if repeated oral administration of different green tea extracts could corroborate the reported hepatotoxicity in humans. In a preliminary 6 week-study, experimental groups (n=9/group) received either the vehicle or a methylene chloride extract (2500 mg/kg body weight) where potential non-polar hepatotoxin(s) could be concentrated. In a second experiment (12 week-study), rats were divided in three groups (n=10/group) and treated with either the vehicle, or an aqueous extract (1400 mg/kg) or AR25 green tea extract (2000 mg/kg). Rat liver functions were assessed by serum biochemistry of hepatotoxicity markers. No sign of evidence of characteristic hepatotoxicity was found in rats treated with very high amount of different green tea extracts in these two experiments (respectively a daily dosage, which was about 900 and 80 times higher to the therapeutic daily dosage of Exolise.     

Effect of ethanol and the alpha-adrenoblockader IEM-611 on the antigenic spectrum of water-soluble brain proteins of rats

The composition of water-soluble antigens in the brain of rats preferring ethanol or water was studied. A significant difference was found in the content of two antigens in rats preferring water or ethanol. Administration of IEM-611 changed the antigen content ot the level characteristic for animals preferring water.     

Effect of ethanol on the metabolism of brain catecholamines

Ethanol, 7 g/kg, was given orally to mice. At various time intervals thereafter the animals received ³H-tyrosine and the net accumulation of ³H-noradrenaline and ³H-dopamine in the brain during 30 min was measured. In some of the experiments the animals were divided into two groups with different degrees of intoxication. In general the accumulation of ³H-catecholamines increased following ethanol, more markedly in the severely than in the less severely intoxicated group. The former group also showed a higher blood ethanol level than the latter. Furthermore, the ratio ³H-dopamine/³H-noradrenaline was significantly increased by ethanol. There were no certain changes in the specific activity of ³H-tyrosine in brain and plasma. The effect of ethanol on the ³H-dopamine accumulation was prevented by nialamide. Endogenous noradrenaline in the brain was reduced by about 15% by ethanol. The only effect of ethanol observed on brain dopamine was a slight and transient decrease.This research was supported by the Swedish State Medical Research Council under grant No. B72-14X-2157-06. The expert technical assistance of Mrs. Barbro AldÄng, Miss Ingrid Blume, Miss Barbro Jörblad and Mrs. Lena Löfberg is gratefully acknowledged.     

Effect of the variations of S-adenosyl-L-methionine liver content on fat accumulation and ethanol metabolism in ethanol-intoxicated rats

The protective effect of S-adenosyl-L-methionine against rat liver steatosis induced by chronic ethanol ingestion was investigated. S-Adenosyl-L-methionine given during ethanol treatment prevented steatosis and accelerated recovery from steatosis when given after ethanol withdrawal. It also caused a slight inhibition of blood ethanol consumption in both acutely and chronically intoxicated rats. About 30% inhibition of alcohol dehydrogenase, but not of the microsomal ethanol oxidation system, occurred in rats subjected to acute ethanol toxicity as well as in normal rats as a consequence of S-adenosyl-L-methionine treatment. A comparison between S-adenosyl-L-methionine and pyrazole, as concerns inhibition of ethanol oxidation and fat accumulation, revealed that a greater inhibition of ethanol metabolism by pyrazole was associated with incomplete prevention of steatosis, while a lower inhibition by S-adenosyl-L-methionine was coupled to a complete prevention. Ethanol induced a drastic decrease of reduced glutathione liver content as well as 630 and 133% increases of blood and liver acetaldehyde contents, respectively. S-Adenosyl-L-methionine treatment almost completely reconstituted the liver reduced glutathione pool and caused a large decrease of the liver and blood acetaldehyde contents. 1-Chloro-2,4-dinitrobenzene, which depletes the cellular reduced glutathione, and diethylethanolamine, an inhibitor of the phosphatidylethanolamine methylation, abolished the S-adenosyl-L-methionine-induced modifications of the reduced glutathione, acetaldehyde, and triacylglycerol contents in the liver of ethanol-treated rats. Neither S-adenosyl-L-methionine nor reduced glutathione inhibitors affected the liver acetaldehyde dehydrogenase activity. It is suggested that, although S-adenosyl-L-methionine induced a small inhibition of ethanol metabolism in the liver, its antisteatosic effect could largely depend on its role as a modulator of the reduced glutathione liver content.     

Effect of cyanamide on the level of endogenous ethanol in the liver of normal rats and in hypocorticism

The rat liver endogenous ethanol level was found to increase under inhibition of aldehyde dehydrogenases by cyanamide. Adrenalectomy results in a decrease of the liver endogenous ethanol content and abolishes cyanamide effect on this index. One of the mechanisms of cyanamide toxic effect may be accumulation of different aldehydes including acetaldehyde.     

Effects of maternal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin on fetal brain growth and motor and behavioral development in offspring rats

The effects of maternal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) during pregnancy on fetal brain growth and neurobehavioral development in early developmental stages were investigated using rat offspring. TCDD in corn-oil (0.1microg/kg) was orally administrated to the dams from the 9th to 19th gestational day. When TCDD effects on the fetal brain weight were analyzed on the 19th gestational day, weight ratio of the brain to the whole body, and that of the forebrain without the cerebral cortex to the whole brain were larger in the exposed group than those of the control group, suggesting premature fetal brain development. TCDD effects on motor functions were investigated using newborns in an inclined plane task. Motor development assessed by righting response on an inclination was delayed in the exposed offspring in the 8th-12th postnatal day, especially in male. Also, TCDD effects on active avoidance behavior in a shuttle box were investigated using the offspring after weaning. Latency in the active avoidance learning was longer, and locomotor activity was reduced in the exposed male offspring in the 41st-44th postnatal day. The results demonstrated that maternal TCDD exposure delayed fetal brain growth and neurodevelopment of the offspring in early stage, especially in male rats.     

Effect of acute ethanol administration on noradrenaline metabolism in brain regions of stressed and nonstressed rats

The effects of ethanol on noradrenaline (NA) metabolism of brain regions in stressed and nonstressed rats were investigated. Male Wistar rats were injected IP with either saline, or ethanol at 0.5 g/kg or 2 g/kg, 5 min before exposure to 1-hr immobilization stress. Levels of NA and its major metabolite, 3-methoxy-4-hydroxyphenylethyleneglycol sulfate (MHPG-SO4) in various brain regions and plasma corticosterone levels were fluorometrically determined. Immobilization stress caused significant increases in MHPG-SO4 levels in all brain regions examined, i.e., the hypothalamus, amygdala, hippocampus, cerebral cortex and locus coeruleus (LC) region. In nonstressed rats, ethanol significantly increased MHPG-SO4 levels in the hypothalamus, hippocampus and cerebral cortex, but not in the amygdala or in the LC region. In stressed rats, ethanol attenuated stress-induced increases in MHPG-SO4 levels preferentially in the amygdala and LC region, but not in the remaining three regions. Although ethanol per se dose-dependently elevated plasma corticosterone levels in nonstressed rats, ethanol at 2 g/kg attenuated the stress-induced elevation of corticosterone. These results suggest that the attenuating effect of ethanol on stress-induced increases in NA turnover in the amygdala and LC region might be related to the stress-relieving properties of this drug.     

Fatty acid alterations in liver and milk of cadmium exposed rats and in brain of their suckling offspring

Fatty acid composition was studied in milk at day 14 and in liver at day 24 after parturition of lactating rats exposed to 0 ppm, 5 ppm or 25 ppm cadmium (Cd) via drinking water for 17 days during lactation, and in the brain of their offspring at day 19 after birth. In the liver phospholipid fraction, 22:5(n-3) was significantly higher, while in the triacylglycerol fraction 22:6(n-3)/20:5(n-3) ratio was significantly lower in the 25 ppm group compared to the controls. Significantly higher proportions of 16:0 and lower proportions of medium-chain fatty acids, 8:0-14:0, were observed in milk of dams in the 25 ppm group, indicating decreased enzymatic activity of thiotransferase II in the mammary gland. Slightly increased levels of 20:3(n-6) were observed in brains of pups in the 25 ppm group compared to control. The results indicate that Cd exposure influences fatty acid metabolism in lactating rats.