Effect of ethanol administration on the in vivo kinetics of thiamine phosphorylation and dephosphorylation in different organs. I. Chronic effects

The effects of chronic ethanol administration on different steps in the metabolism of thiamine (T), thiamine mono- (TMP) and pyrophosphate (TPP) were determined in vivo in the liver, kidney, heart, skeletal muscle and small intestinal mucosa. The radioactivity of T and its phosphoesters was measured in plasma and in the selected organs under steady-state conditions and at fixed time intervals (0.25-240 hr) after an i.p. injection of Thiazole-[2-14C]-thiamine (30 micrograms: 1.25 microCi) in rats chronically (35 days) ethanol-treated (daily dose of 4.7 g kg-1 body wt by gastric gavage). Two types of controls were used: pair-fed rats treated with a sucrose solution isoenergetic with ethanol, and water-treated rats. A nutritionally adequate diet, which supplied an excess of thiamine, was given to the rats, producing a virtually steady content of thiamine compounds in the tissues. The analytical data obtained were elaborated using appropriate compartmental mathematical models, which allowed the fractional rate constants, turnover rates and turnover times to be calculated. Alterations in thiamine metabolism were modest and differed according to the organs. The most widespread modification was to facilitate the entry of T (small intestine, kidney and heart) or TMP (small intestine and kidney), while no significant change of T and TMP release was seen. Sucrose had minimal effect in both steps. Enzymatic transformations of thiamine were likewise marginally affected. A general trend toward a slower T pyrophosphorylation and a faster T phosphate dephosphorylation was observed in the small intestine, kidney, heart and liver. Skeletal muscle was unaffected.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of dietary zinc deficiency on the endogenous phosphorylation and dephosphorylation of rat erythrocyte membrane

The effect of dietary zinc deficiency on patterns of phosphorylation and dephosphorylation of rat erythrocyte membrane proteins and erythrocyte filterability was examined. Weanling male Wistar rats were fed an egg white-based diet containing less than 1.1 mg zinc/kg diet ad libitum for 3 wk. Control rats were either pair-fed or ad libitum-fed the basal diet supplemented with 100 mg zinc/kg diet. Net phosphorylation and dephosphorylation of erythrocyte membrane proteins were carried out by an in vitro assay utilizing [gamma-32P]ATP. The membrane proteins were subsequently separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the 32P content of gel slices was counted by Cerenkov counting. Erythrocyte filterability was measured as the filtration time of suspensions of erythrocytes, both untreated and preincubated with diamide, under constant pressure. Erythrocyte ghosts from zinc-deficient rats demonstrated greater dephosphorylation of protein bands R1 plus R2 and R7 than pair-fed rats and greater net phosphorylation of band R2.2 than pair-fed or ad libitum-fed control rats (P less than 0.05). Erythrocytes from ad libitum-fed control rats showed significantly longer filtration times than those from zinc-deficient or pair-fed control rats. In conclusion, dietary zinc deficiency alters in vitro patterns of erythrocyte membrane protein phosphorylation and dephosphorylation, whereas the depression in food intake associated with the zinc deficiency increases erythrocyte filterability.     

Effects of acute and chronic ethanol administration on thiamine metabolizing enzymes in some brain areas and in other organs of the rat

The effect of ethanol (4.7 g/kg body wt intragastrically as a single dose or once daily for 35 days) on the levels of the thiamine metabolizing enzymes (thiamine pyrophosphokinase, TPKase; thiamine pyrophosphatase, TPPase; and monophosphatase, TMPase) was studied in different organs (liver, kidney, small intestine, heart and skeletal muscle) and nervous regions (cerebral cortex, cerebellum, medulla oblongata, pons, corpus callosum, hypothalamus and sciatic nerve) of the rat. In order to evaluate the non-specific effects of the stress of gastric gavage and of the additional caloric intake, appropriate control groups of animals were treated intragastrically with water or with a saccharose solution isoenergetic with ethanol respectively. All animals were reared on a nutritionally adequate diet supplying amounts of thiamine higher than the recommended daily requirement. Enzymatic activities were determined quantitatively by biochemical methods. Tissue TPKase levels were generally reduced by both acute and chronic ethanol administration. TPPase levels were generally reduced after acute and increased after chronic ethanol treatment. Changes in brain TMPase levels were similar to those observed for TPPase. In visceral organs and skeletal muscle TMPase activity was increased by chronic ethanol treatment as compared to acute ethanol administration. In conclusion, ethanol exerts a marked influence on the tissue levels of the thiamine metabolizing enzymes: the activity of the enzymes dephosphorylating thiamine phosphates is increased whereas the activity of the thiamine pyrophosphate synthesizing enzyme is reduced. These changes may contribute to an important extent to the disturbances in thiamine cellular uptake and metabolism observed in alcoholism.     

Acute and chronic effects of ethanol on learning-related synaptic plasticity

Alcoholism is associated with acute and long-term cognitive dysfunction including memory impairment, resulting in substantial disability and cost to society. Thus, understanding how ethanol impairs cognition is essential for developing treatment strategies to dampen its adverse impact. Memory processing is thought to involve persistent, use-dependent changes in synaptic transmission, and ethanol alters the activity of multiple signaling molecules involved in synaptic processing, including modulation of the glutamate and gamma-aminobutyric acid (GABA) transmitter systems that mediate most fast excitatory and inhibitory transmission in the brain. Effects on glutamate and GABA receptors contribute to ethanol-induced changes in long-term potentiation (LTP) and long-term depression (LTD), forms of synaptic plasticity thought to underlie memory acquisition. In this paper, we review the effects of ethanol on learning-related forms of synaptic plasticity with emphasis on changes observed in the hippocampus, a brain region that is critical for encoding contextual and episodic memories. We also include studies in other brain regions as they pertain to altered cognitive and mental function. Comparison of effects in the hippocampus to other brain regions is instructive for understanding the complexities of ethanol's acute and long-term pharmacological consequences.     

Effect of thiamine deprivation on thiamine metabolism in mice

Mice were made deficient by thiamin deprivation. Thiamin pyrophosphokinase and thiamin pyrophosphatase activities were measured, and their changes were compared to transketolase (TK), pyruvate and alpha-ketoglutarate dehydrogenase (DG) activities and thiamin pyrophosphate (TPP) level in the same tissues. Thiamin pyrophosphokinase activity was increased on day 10 of thiamin deficiency and remained unchanged within the next period of investigation. Thiamin pyrophosphatase activity decreased after 5 days and was enhanced in 10, 15 and 20 days of deficiency. This fact may be related to the regulatory role of the enzyme in the intertissue vitamin distribution and transport. The liver TPP level was the earliest and most specific indicator of the thiamin status as thiamin deficiency developed, and pyruvate DG was more drastically inhibited than alpha-ketoglutarate DG. Blood TK was more sensitive to thiamin deprivation than liver TK. In both cases we obtained a more (20-40% in blood) or less (5-10% in liver) pronounced TPP-stimulatory effect of transketolase activity.     

The acute and chronic effects of ethanol on cardiac muscle protein synthesis in the rat in vivo.

An investigation was made into the acute and chronic effects of ethanol on rates of protein synthesis in the hearts of young rats (80-100 g body weight). Acute ethanol administration (75 mmol/kg body weight, IP) significantly reduced the fractional rate of protein synthesis by 20% after 2.5 hr, compared with saline-treated controls. Chronic ethanol feeding (36% of total calories) for 6 weeks significantly reduced cardiac wet weight by 11%, when compared to rats fed isovolumetric amounts of the same diet in which ethanol was substituted by isocaloric glucose. Neither the concentration nor the content of mixed cardiac proteins relative to body weight were overtly altered by chronic ethanol feeding, although, the total content of mixed cardiac proteins were significantly decreased. RNA concentrations and RNA relative to body weight increased slightly, but total cardiac DNA decreased. Indices for the capacity or potential of the heart to synthesis protein (indicated by the RNA/protein and RNA/DNA ratios) and the "DNA-unit" (protein/DNA ratio) were increased in response to chronic ethanol treatment. The fractional and absolute rates of mixed protein synthesis in the heart were (relatively) unaltered by chronic ethanol treatment, as was RNA efficiency and synthesis relative to DNA. It was concluded that the heart displays contrasting responses to acute and chronic ethanol exposure.     

Pre-natal and post-natal effects of alcohol in the rat--II. Changes in gamma-aminobutyric acid concentration and adenosine triphosphatase activity in the brain

The effects of chronic alcohol administration during the pre-nataland/or post-natal period on concentration of aminobutyric acid(GABA) in the amygdaloid cortex and striatum and on activitiesof Na⁺, K⁺- and Mg²⁺ -activated adenosine triphosphatases (ATPases)in the mid-brain and hippocampus were studied. There was a smalldecrease in the GABA content of both brain regions in thoseoffspring that had received alcohol immediately before, butnot in those that were free of the drug at the time of death.This suggests that the changes in GABA concentration are dueto a direct pharmacological effect of ethanol. A decrease alsooccurred in the activity of the Mg²⁺ -activated ATPase in thoseoffspring exposed to alcohol during the period of weaning. Thiseffect could not be attributed to a direct action of the drug,but may be indicative of a longer-term influence of ethanolon membrane transport processes. The increase in the activityof the Na⁺, K⁺ -activated ATPase in the mid-brain of rats thatwere exposed to the drug pre-natally and post-natally and followingweaning is probably due to a direct effect of the drug.     

The effects of maternal thiamine nutrition on thiamine status of the offspring in broiler chickens

The response of broiler chickens to a wide range of dietary supplementation of thiamine to broiler breeder diet was studied in order to understand the effects of maternal thiamine nutrition on the status of thiamine indices in the offspring. Thiamine, and thiamine pyrophosphate (TPP) content, and alpha-ketoglutarate dehydrogenase (KGDH) activity were measured in hearts from 20 day old chicken embryos and from chickens at 1, 7, 14, and 21 days of age and in blood at 21 days of age. Total thiamine content in the heart of day old chicks was higher in comparison to 20 day old embryos. Maternal supplementation of thiamine increased heart thiamine in the offspring (p < 0.001), and increased the activity of KGDH in the hearts of day old chicks (p < 0.001), but not in the embryo. The TPP content in the heart increased in response to both maternal and offspring thiamine supplementation (p < 0.001), however the effect of broiler thiamine supplementation was largely independent from the maternal effect. The effect of maternal thiamine nutrition on the offspring's heart KGDH activity was apparent, but the responses to broiler supplementation were dependent largely on the maternal effect. Blood TPP content was not affected by maternal thiamine supplementation (p = 0.39), but thiamine supplementation in the offspring diets increased blood TPP (p < 0.001). Both maternal and offspring thiamine supplementation increased blood free base thiamine content (both p < 0.001). It is concluded from this study that maternal thiamine nutrition affects thiamine status indices and thiamine metabolism of the offspring.     

Combined effects of ethanol and garlic on hepatic ethanol metabolism in mice.

The combined effects of ethanol and components in fresh garlic on ethanol metabolism were investigated in the livers of mice. Male, 11-wk-old C3H/HeNCrj mice were intragastrically administered 2 g ethanol/kg body weight after being administered fresh garlic juice for 8 d (garlic group), and changes in the concentrations of ethanol, acetaldehyde and acetate in the serum, and changes in the activity of hepatic enzymes related to ethanol metabolism in mice were examined. The increases in the concentrations of acetaldehyde and acetate in the serum after ethanol administration tended to be diminished following garlic administration. The microsomal ethanol-oxidizing system (MEOS) in the livers of the garlic groups was significantly lower than that of the control microsomes at 2 h after ethanol administration. It therefore seems that the decrease of MEOS in hepatic microsomes caused a smaller increase in the acetaldehyde concentration in the serum of the garlic groups because cytosolic alcohol dehydrogenase showed no significant difference between the control and garlic groups. After ethanol administration, the content of cytochrome P-450 in the hepatic microsomes of the control groups increased, while that of the garlic groups did not change although cytochrome P-450 (CYP) 2E1 and 1A2 in the hepatic microsomes of the garlic groups increased. These results indicate that the induction of isozymes of cytochrome P-450 other than CYP 2E1 and 1A2 was inhibited following garlic administration. Cytosolic high Km and total aldehyde dehydrogenase (AIDH) in the liver of the garlic groups tended to be lower than those activities of the control groups at 1 and 2 h after ethanol administration. It therefore seems that the decreases of AIDH in the hepatic cytosols diminished the increase of acetate in the serum of the garlic groups after ethanol administration. These results suggest that the ethanol metabolism in the mouse liver is controlled by components in fresh garlic juice.     

Effects of ethanol on the kinetics of methyl ethyl ketone in man.

The kinetics of inhaled methyl ethyl ketone (MEK) at a concentration of 200 ppm for four hours were studied in volunteers after swallowing ethanol at a dose of 0.8 g/kg. Ethanol was given either before or at the end of the exposure to MEK. The blood concentrations of MEK, 2-butanol, and 2,3-butanediol were monitored during and after the exposure. MEK concentrations in exhaled air and MEK and 2,3-butanediol concentrations in urine were also measured. Ethanol inhibited the primary oxidative metabolism of MEK and caused an increase in the blood concentrations of MEK and 2-butanol after ingestion. Ethanol ingestion, through higher blood MEK concentrations, also increased the elimination of MEK in the urine and exhaled air. Ethanol taken before exposure to MEK reduced the serum concentration of 2,3-butanediol initially but there was an increase about eight hours after the exposure. Urinary excretion of 2,3-butanediol followed the same pattern. Prior ingestion of ethanol thus seemed to interfere with the metabolism of 2,3-butanediol during and after exposure to MEK.     

Effect of ascorbic Acid and thiamine supplementation at different concentrations on lead toxicity in liver

OBJECTIVE: To investigate the effect of ascorbic acid [vitamin C (VC)] on liver damage parameters in the lead-exposed mice, when given in combination with thiamine [vitamin B1 (VB(1))] at different concentrations. METHODS: Sixty-six male mice were randomly assigned into 11 groups (n = 6). Mice in Group I were supplied with only the tap water as the drinking water; mice in Group II were provided with a tap water containing 0.2% lead acetate; mice in Group III-XI were given different dose of VC (140, 420, 1260 mg kg(-1) bw) and VB(1) (10, 30, 90 mg kg(-1) bw) according to 3 x 3 factorial design by oral gavages, along with ingestion of 0.2% lead acetate. After 42 test days, DNA damage of liver cells was assessed using single-cell gel electrophoresis. The apoptosis rate of liver cells was determined by flow cytometry. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in blood and the level of reduced glutathione (GSH) in liver cells were measured based on individual biochemical reactions. RESULTS: Compared with the Group I, sub-chronic lead ingestion (Group II) resulted in a significant decrease of Hb, GSH-P(X), SOD in blood and GSH level in liver cells; lead exposure induced also a significant increase in DNA damage and apoptosis of liver cells (P < 0.05). Supplementation with VC and VB(1), however, reversed these effects. The best effective combination was VC (420 mg kg(-1) bw) and VB(1) (10 mg kg(-1) bw), followed by the combination of VC (420 mg kg(-1) bw) and VB(1) (30 mg kg(-1) bw). But no reversion was shown in the combination of the highest combination of VC (1260 mg kg(-1)) and VB(1) (90 mg kg(-1)). CONCLUSIONS: These findings strongly indicated that combination of VC and VB(1) can lessen the damage to liver cells from oxidative stress induce by lead, but the antioxidant effects are dependent on their concentrations.     

Acute ethanol administration: effects on stress-induced gastric and duodenal ulcer in rats

Rats were given 6% ethanol (v/v) as their only source of liquid for 4 days. On the basis of ethanol consumption (g/kg/day), animals were divided into high, medium and low ethanol consuming groups. A non-ethanol exposed control group was also included. Following a 24 hr food deprivation period, animals were restrained for 3 hr. No differences in gastric ulcer frequency or severity were noted with the exception of a slight tendency toward a lower incidence among ethanol consuming rats relative to controls. An unusual observation was the high incidence of duodenal ulcer observed only among ethanol consuming rats. This ethanol-stress interaction is discussed in terms of an animal's history of ethanol exposure.     

Acute ethanol intoxication during pregnancy: postnatal effects on the behavioral response to serotonin agents.

Pregnant wistar rats were treated on the eighth day of gestation (GD 8) with two IP injections, spaced by 4 h, of either ethanol (2.9 g/kg in 24% v/v saline solution, EG) or saline (SG). Other pregnant females did not received any type of IP injections (absolute control group, ACG). Offspring were tested at 45 or 90 days of age. At 45 days of age, EG showed an increased behavioral response (forepaw treading and hindlimb abduction) to the 5-HT1 agonist, 5-methoxy-N,N-dymethyltryptamine. In addition, an enhanced "wet-dog" shakes behavioral response to 5-HT2 agonist, 5-hydroxy-L-tryptophan, was also observed in EG as compared to ACG and SG. On the contrary, at 90 days, EG exhibited a diminished behavioral reactivity to 5-HT1 and 5-HT2 agonists as compared to SG. These results demonstrated that acute administration of ethanol on GD 8 induced long-lasting changes in the functioning of central serotonergic systems.     

不同腹腔减压方式对腹腔间室综合征大鼠肠道功能的影响

目的:观察腹腔开放减压和腹腔梯度减压两种方式对腹腔间室综合征(ACS)大鼠肠黏膜屏障的影响. 方法:将18只大鼠模型随机分为对照组、开放减压组和梯度减压组.其中开放减压组大鼠ACS持续4h后,经腹正中切口迅速使腹腔压力(IAP)降至0mmHg;梯度减压组大鼠ACS持续4h后,通过减慢气腹充气速度,以ΔIAP=-0.5 mmHg/min的速度减压,直至IAP恢复至0 mmHg(减压时间为50 min).在IAP降至0 mmHg后4h处死动物,取末端回肠行组织病理学检查、肠系膜淋巴结细菌易位测定、肠黏膜组织肿瘤坏死因子α(TNF-α)、白细胞介素-6(IL-6)和肠型脂肪酸结合蛋白(i-FABP)测定. 结果:梯度减压组与开放减压组比较,肠道病理损伤更小,细菌易位率更低,炎性因子水平更低.其中肠道通透性、细菌易位菌落数、IL-6水平在不同减压方式组之间差异有统计学意义(P＜0.05). 结论:不同的腹腔减压方式对ACS大鼠的肠道功能影响不同.梯度腹腔减压比开放减压对ACS大鼠肠道功能的损伤更小.     

Effect of thiamine deficiency, pyrithiamine and oxythiamine on pyruvate metabolism in rat liver and brain in vivo

Rats were fed either a thiamine-deficient diet of diets containing pyrithiamine or oxythiamine. When symptoms of thiamine deficiency appeared, the animals were injected intraperitoneally with [2-14C] pyruvate six to twelve minutes prior to sacrifice. Free glutamic and aspartic acids were isolated from liver and brain and degraded. The results indicate that, in thiamine-deficient or oxythiamine-treated rats, pyruvate metabolism in liver and brain is similar to that in normal animals. In contrast, pyrithiamine drastically decreases the oxidative decarboxylation of pyruvate by rat liver.     

Acute effects of ethanol on cultured myocardial cells: an ultrastructural study

Myocardial cells in monolayer culture were examined ultrastructurally at various times (5 min-24 hr) after addition of ethanol (12.5-500 mM). At 5 min, all concentrations of ethanol induced mitochondrial fusion and a remarkable increase in glycogen granules, and the mitochondrial outer membrane seemed more fragile. These initial ultrastructural changes then recovered with time. Glycogen granules were decreased, then returned to normal more rapidly as the ethanol concentration was increased. Mitochondrial fusion and fragile appearance of the outer membrane were hardly seen at 1 hr in the presence of 12.5 or 50 mM ethanol, but were still evident with higher ethanol concentrations (200 and 500 mM), though less frequently than at 5 and 30 min. Furthermore, 'giant mitochondria' (with at least a five-fold increase in sectional area) appeared in the presence of 200 and 500 mM ethanol with increasing time of exposure. These results are discussed in relation to the direct effects of ethanol.     

Volume and dose effects of experimenter-administered ethanol preloads on ethanol seeking and self-administration.

The present experiment used a behavioral model developed to separate the initial behavior required to obtain access to ethanol (appetitive responding or lever presses) from the actual self-administration (consummatory responding or intake) to test the hypothesis that these responses are under the control of different behavioral/physiological processes, and therefore differentially affected by an ethanol priming dose. In male, Long Evans rats, "preload" volume (0.5 and 2.0ml) and dose (approximately 10%, 25%, and 50% of the total normally consumed in nontreatment sessions translating to 0.1, 0.25, and 0.5g/kg) of ethanol were varied and administered by the experimenter via oral gavage prior to an operant session. Overall, there were no priming effects, or increases, in ethanol-reinforced responding resulting from the ethanol preloads. The findings showed that the low preload volume produced linear, dose-dependent decreases in both intake and seeking. However, while the high volume also produced a linear dose-dependent decrease in ethanol seeking, there was a decrease in intake at every dose. That is, ethanol seeking was insensitive to preload volume, while intake was affected in a dose-dependent manner except at the lowest dose when preload volume did play a role in intake regulation. These findings indicate that "fullness" and pharmacological cues differentially impact the appetitive and consummatory behaviors reinforced by ethanol solutions, with intake being more sensitive to preload volume and seeking being more sensitive to preload pharmacology.