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Developmental changes in the modulation of synaptic glycine receptors by ethanol

JOURNAL OF NEUROPHYSIOLOGY 2000年第5期84卷 2409-2416页

作者： Eggers, ED O'Brien, JA Berger, AJ Univ Washington Sch Med Dept Physiol & Biophys Seattle WA 98195 USA

During postnatal motoneuron development, the glycine receptor (GlyR) alpha subunit changes from alpha2 (fetal) to alpha1 (adult). To study the effect this change has on ethanol potentiation of GlyR currents in hypoglossal motoneurons (HMs), we placed neurons into two groups: neonate [postnatal day 1 to 3 (P1-3)], primarily expressing a2, and juvenile (P9-13), primarily expressing alpha1. We found that glycinergic spontaneous miniature inhibitory postsynaptic currents (mIPSCs) in neonate HMs are less sensitive to ethanol than in juveniles. Thirty millimolar ethanol increased the amplitude of juvenile mIPSCs but did not significantly change neonatal mIPSCs. However, 100 mM ethanol increased the amplitudes of both neonate and juvenile mIPSCs. There was a significant difference between age groups in the average ethanol-induced increase in mIPSC amplitude for 10, 30, 50, and 100 mM ethanol. In both age groups ethanol increased the frequency of glycinergic mIPSCs, but there was no difference in the amount of frequency increase between age groups. Ethanol (100 mM) also potentiated evoked IPSCs (eIPSCs) in both neonate and juvenile HMs. As we observed for mIPSCs, 30 mM ethanol increased the amplitude of juvenile eIPSCs, but had no significant effect on eIPSCs in neonate HMs. Ethanol also potentiated currents induced by exogenously applied glycine in both neonate and juvenile HMs. These results suggest that ethanol directly modulates the GlyR. To investigate possible mechanisms for this, we analyzed the time course of mIPSCs and single-channel conductance of the GlyR in the presence and absence of ethanol. We found that ethanol did not significantly change the time course of mIPSCs. We also determined that ethanol did not significantly change the single-channel conductance of synaptic GlyRs, as estimated by nonstationary noise analysis of mIPSCs. We conclude that the adult form of the native GlyR is more sensitive to ethanol than the fetal form. Further, enhancement of GlyR

关键词：年龄因素动物新生脑干/细胞学脑干/生长和发育脑干/生理学中枢神经系统抑制药/药理学导电性乙醇/药理学诱发电位/药物作用诱发电位/生理学甘氨酸/生理学膜电位/药物作用膜电位/生理学神经抑制/药物作用神经抑制/生理学大鼠 Sprague-Dawley 反应时间/药物作用反应时间/生理学受体甘氨酸/代谢突触/药物作用突触/代谢动物大鼠

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In vitro effects of ethanol, acetaldehyde and fatty acid ethyl esters on human erythrocytes

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ALCOHOL AND ALCOHOLISM 2002年第2期37卷 179-186页

作者： Tyulina, OV Prokopieva, VD Dodd, RD Hawkins, JR Clay, SW Wilson, DO Boldyrev, AA Johnson, P Ohio Univ Dept Biomed Sci Athens OH 45701 USA Ohio Univ Dept Chem & Biochem Athens OH 45701 USA Ohio Univ Dept Geriatr Med Athens OH 45701 USA Russian Acad Med Sci Mental Hlth Res Inst Tomsk Russia Moscow MV Lomonosov State Univ Ctr Mol Med Moscow 119899 Russia Moscow MV Lomonosov State Univ Int Biotechnol Ctr Moscow 119899 Russia

In vitro experiments were performed to determine if ethanol was metabolized by human erythrocytes and to investigate if ethanol or its metabolites, acetaldehyde and fatty acid ethyl esters, affected erythrocyte morphology and stability. No detectable metabolism of ethanol was found in erythrocytes, although ethanol itself caused an elevated rate of spontaneous haemolysis in erythrocyte preparations, Physiologically attainable levels of ethanol were found to stabilize erythrocytes against haemolysis induced by sodium hypochlorite, and the presence of ethanol caused a decrease in erythrocyte reactive oxygen species levels, although the mechanism for such a process is unknown. Both physiologically attainable. and higher levels of acetaldehyde had no effects on erythrocyte morphology and stability even after a 16 h exposure. Fatty acid ethyl esters caused structural changes and instability in erythrocytes in vitro, but whether such changes occur in vivo has not been established. The results of these studies suggest that the deleterious effects of ethanol consumption on erythrocytes in vivo may be. at least in part, the result of direct effects of unmetabolized ethanol on erythrocyte components.

关键词：乙醛/代谢乙醛/药理学过氧化氢酶/拮抗剂和抑制剂中枢神经系统抑制药/代谢中枢神经系统抑制药/药理学酶激活/药物作用红细胞/细胞学红细胞/药物作用红细胞/酶学酯类/代谢酯类/药理学乙醇/代谢乙醇/药理学脂肪酸类/代谢脂肪酸类/药理学自由基/代谢溶血/药物作用溶血/生理学亚麻酸类/药理学人类男(雄)性

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Chronic intragastric infusion of ethanol-containing diets induces CYP3A9 while decreasing CYP3A2 in male rats

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JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS 2000年第2期295卷 747-752页

作者： Rowlands, JC Wang, HM Hakkak, R Ronis, MJJ Strobel, HW Badger, TM Univ Arkansas Med Sci Dept Pediat Little Rock AR 72202 USA Univ Arkansas Med Sci Arkansas Childrens Hosp Res Inst Little Rock AR 72202 USA Univ Texas Sch Med Dept Biochem & Mol Biol Houston TX USA

The CYP3A subfamily is the most abundant of the human hepatic cytochrome P450 enzymes. They mediate the biotransformation of many drugs, including a number of psychotropic, cardiac, analgesic, hormonal, immunosuppressant, antineoplastic, and antihistaminic agents. We studied diet/ethanol interactions using total enteral nutrition in adult male Sprague-Dawley rats with diets containing 16% protein, ethanol (13 g/kg), corn oil (fat;25-45%), and carbohydrate (CHO;1-21%). Using this model, chronic ethanol feeding decreased CYP3A activity (testosterone 6 beta -hydroxylation) and apoprotein levels (Western blot) (P<.05) and these effects were independent of the dietary CHO/fat ratio. The CYP3A2 mRNA levels decreased (P<.05) in the rats fed ethanol-containing diets by 73 to 83% compared with rats fed control diets, regardless of the CHO/fat ratio. In contrast, ethanol induced CYP3A9 mRNA levels (P<.05) and this effect was greater (P<.05) in the high-CHO/lowfat group (11.3-fold) than in the low-CHO/high-fat group (2.6-fold). Purified recombinant rat P450 3A9 had a chlorzoxazone 6-hydroxylase activity with a turnover number 1.3 nmol/min/nmol of P450. These results indicate that 1) ethanol differentially affects the expression of CYP3A gene family and this regulation appears to be modulated by dietary CHO/fat ratio;2) the decrease in testosterone 6 beta -hydroxylase activity and CYP3A apoprotein levels are most likely due to the ethanol-induced decrease in CYP3A2 mRNA levels;and 3) CYP3A9 is induced by ethanol and is a low-affinity, high-K-m chlorzoxazone hydroxylase.

关键词：芳基烃羟化酶类中枢神经系统抑制药/药理学氯唑沙宗/药理学细胞色素P-450CYP3A 细胞色素P450酶系统/生物合成细胞色素P450酶系统/遗传学细胞色素P450酶系统/代谢膳食膳食碳水化合物/药理学膳食脂肪类/药理学肠道营养酶诱导/药物作用乙醇/药理学同工酶类/生物合成同工酶类/遗传学同工酶类/代谢氧化还原酶类 N-脱甲基/生物合成氧化还原酶类 N-脱甲基/遗传学氧化还原酶类 N-脱甲基/代谢 RNA 信使/遗传学 RNA 信使/代谢大鼠 Sprague-Dawley 甾类羟化酶类/生物合成甾类羟化酶类/代谢底物特异性动物男(雄)性大鼠

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Cocaine and alcohol interactions in the rat: Effect on cocaine pharmacokinetics and pharmacodynamics

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JOURNAL OF PHARMACEUTICAL SCIENCES 1999年第4期88卷 459-467页

作者： Pan, WJ Hedaya, MA Washington State Univ Coll Pharm Dept Pharmaceut Sci Pharmacol Toxicol Grad Program Pullman WA 99164 USA

The effect of alcohol coadministration on cocaine pharmacokinetics and pharmacodynamics was investigated in awake, freely moving rats. Cocaine plasma and brain extracellular fluid (ECF) concentration-time profiles were characterized after intraperitoneal (ip) administration of 30 mg/kg cocaine to rats that were pretreated with either normal saline or alcohol at 5 g/kg in a balanced crossover experimental design. The neurochemical response to cocaine administration, measured as the change in dopamine concentration in the nucleus accumbens (N ACC) and the change in the mean arterial blood pressure were monitored simultaneously. Intragastric alcohol administration significantly increased cocaine systemic bioavailability after ip administration from 0.550 +/- 0.044 to 0.754 +/- 0.071. Also, the absorption rate constant increased from 0.199 +/- 0.045 to 0.276 +/- 0.059 min(-1) due to alcohol coadministration;however, this increase was not significant. Alcohol inhibition of cocaine metabolism caused an increase in cocaine elimination half-life from 26.3 +/- 3.6 to 40.0 +/- 8.1 min. Also, cocaine tissue distribution was enhanced by alcohol, resulting in a significant increase in cocaine Volume of distribution. Analysis of the brain cocaine concentration-neurochemical effect relationship by the sigmoid-E-max pharmacodynamic model showed that E-max increased from 850 +/- 200 to 1550 +/- 640% of baseline due to alcohol coadministration, whereas EC50 decreased from 3400 +/- 580 to 2000 +/- 650 ng/mL, indicating higher cocaine potency in the presence of alcohol. The estimates of the indirect inhibitory pharmacodynamic model used to examine the plasma cocaine concentration-change in blood pressure relationship were not significantly different after the two treatments. These results indicate that alcohol significantly alters cocaine absorption, distribution, and elimination, resulting in higher and prolonged cocaine plasma concentration. Alcohol coadministration also potentiates

关键词：算法生物转化血压/药物作用脑化学/药物作用中枢神经系统抑制药/血液中枢神经系统抑制药/药理学可卡因/药代动力学可卡因/药理学多巴胺/代谢多巴胺摄取抑制剂/药代动力学多巴胺摄取抑制剂/药理学药物相互作用乙醇/血液乙醇/药理学注射腹腔内注射静脉内大鼠 Wistar 动物男(雄)性大鼠

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Interactive role for neurosteroids in ethanol enhancement of γ-aminobutyric acid-gated currents from dissociated substantia nigra reticulata neurons

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JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS 1999年第3期291卷 1054-1059页

作者： Criswell, HE McCown, TJ Ming, Z Mueller, RA Breese, GR Univ N Carolina Sch Med Bowles Ctr Alcohol Studies Chapel Hill NC 27599 USA Univ N Carolina Ctr Neurosci Dept Psychiat Chapel Hill NC USA Univ N Carolina Ctr Neurosci Dept Anesthesiol Chapel Hill NC USA Univ N Carolina Ctr Neurosci Dept Pharmacol Chapel Hill NC USA Univ N Carolina Sch Med Gene Therapy Ctr Chapel Hill NC 27599 USA

Although previous in vivo electrophysiological studies demonstrated a consistent ethanol enhancement of gamma-aminobutyric acid (GABA) responsiveness from substantia nigra reticulata (SNR) neurons, ethanol applied in vitro to dissociated neurons from the SNR had an inconsistent effect on GABA function. One source for the disparity between these contrasting in vivo and in vitro results could be an endogenous factor (acting on an auxiliary site on GABA(A) receptors) that was not available to the isolated SNR neurons. Because neurosteroids are present in vivo and act on an auxiliary site, it was hypothesized that the presence of a neurosteroid was important for a consistent effect of ethanol on GABA responsiveness from neurons studied in vitro. Alone, the neurosteroid analog alphaxalone produced a significant, concentration-related enhancement of GABA responsiveness from isolated SNR neurons. In contrast to an inconsistent action of 100 mM ethanol on GABA responsiveness in the absence of alphaxalone, the presence of 30 and 100 nM alphaxalone resulted in the majority of isolated neurons responding to this ethanol level. At a concentration of alphaxalone as low as 30 nM, ethanol produced a robust concentration- related increase in GABA-gated currents from this cell type. The neurosteroid 3 alpha,5 alpha-tetrahydrodeoxycorticosterone (100 nM) also permitted a reliable concentration- dependent ethanol enhancement of responses to GABA from SNR cells, indicative that the effects of alphaxalone were not unique. This consistent neurosteroid-induced ethanol enhancement of GABA responsiveness from dissociated SNR neurons supports the view that neurosteroids may play a key role in the action of ethanol on postsynaptic GABA(A) receptor function.

关键词：麻醉药/药理学细胞培养的中枢神经系统抑制药/药理学脱氧皮质酮/类似物和衍生物脱氧皮质酮/药理学电生理学乙醇/药理学离子通道闸门/药物作用神经元/药物作用神经元/代谢膜片钳术孕烷二酮类/药理学甾类/药理学黑质/细胞学黑质/药物作用黑质/代谢 γ氨基丁酸/生理学动物大鼠

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C-fos and egr-1 immediate-early gene induction by cocaine and cocaethylene in rat brain -: A comparative study

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NEUROBIOLOGICAL MECHANISMS OF DRUGS OF ABUSE: COCAINE, IBOGAINE, AND SUBSTITUTED AMPHETAMINES 2000年第1期914卷 46-57页

作者： Thiriet, N Aunis, D Zwiller, J Unite INSERM U338 Ctr Neurochim F-67084 Strasbourg France

The induction of immediate-early genes can now be considered as a tool to study neuronal activation in different brain structures. These genes, which are rapidly and transiently induced in response to diverse extracellular stimulation, coordinate alterations in gene expression underlying neuronal plasticity. Using in situ hybridization, we found that acute i.p. cocaine (20 mg/kg) injection produced a strong expression of egr-1 and c-fos genes in the nucleus accumbens, caudate-putamen, and frontal cortex in the rat. Cocaethylene is an active metabolite of cocaine that is formed when cocaine is consumed together with ethyl alcohol. Injection of cocaethylene at a dose equivalent to cocaine induced the expression of the two immediate-early genes in the same brain structures, but to a lesser extent. A high dose of ethanol increased egr-1 and c-fos expression in the frontal cortex and in the lateral part of the caudate-putamen. Since cocaine is known to potently inhibit both dopamine and serotonin transporters, whereas cocaethylene only inhibits the dopamine transporter, our results strongly suggest that the serotonergic system participates in the mode of action of cocaine in its ability to trigger immediate-early gene transcription.

关键词：放射自显影术/方法脑/解剖学和组织学脑/药物作用中枢神经系统抑制药/药理学可卡因/类似物和衍生物可卡因/药理学多巴胺摄取抑制剂/药理学早期生长反应蛋白质1/遗传学早期生长反应蛋白质1/代谢乙醇/药理学基因表达调控/药物作用原位杂交/方法原癌基因蛋白质c-fos/遗传学原癌基因蛋白质c-fos/代谢大鼠 Wistar 转录激活动物男(雄)性大鼠

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Prolonged anesthetic recovery

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CLINICAL TECHNIQUES IN SMALL ANIMAL PRACTICE 1999年第1期14卷 56-64页

作者： Broadstone, RV Virginia Polytech Inst & State Univ Virginia Maryland Reg Coll Vet Med Dept Small Anim Clin Sci Blacksburg VA 24061 USA

Failure to awaken, delayed awakening, and blunted responsiveness following anesthesia constitute a continuum which must be recognized, diagnosed accurately, and responded to appropriately. One must determine whether it is a normal recovery for this animal or a pathological delay. A delayed recovery results in continued depression of many organ systems. The possible etiologic factors, some obvious, others quite obscure, must be considered quickly to provide the patient with the best possible care. Diminished responsiveness is usually the result of a complex interaction between multiple factors. A systematic approach to treatment first begins by ensuring cardiac output, oxygenation, ventilation, and supportive care in all patients. Then consider the potential causes for prolonged recovery and either rule them out or treat them. First consider the potential causes for central nervous system depression. Several commonly encountered medical conditions enhance sensitivity to or decrease the elimination of anesthetic and sedative agents. The astute clinician will be aware of the medical condition and its impact on anesthetic drug effect and elimination to avoid prolonged recoveries.

关键词：麻醉/兽医学麻醉恢复期动物疾病/外科学中枢神经系统抑制药/药理学兽医学动物

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A longitudinal study of the effects of prenatal ethanol exposure on neuronal acquisition and death in the principal sensory nucleus of the trigeminal nerve: interaction with changes induced by transection of the infraorbital nerve

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JOURNAL OF NEUROCYTOLOGY 1999年第12期28卷 999-1015页

作者： Miller, MW Vet Affairs Med Ctr Res Serv Iowa City IA 52246 USA Univ Iowa Coll Med Dept Psychiat Iowa City IA 52242 USA Univ Iowa Coll Med Dept Pharmacol Iowa City IA 52242 USA

The present study determines (1) whether ethanol-induced microencephaly results from reductions in neuronal acquisition (i.e., cell proliferation and neuronal migration) and/or increases in neuronal death and (2) whether ethanol exacerbates death by the same mode as that for naturally occurring or lesion-induced neuronal death. Pregnant rats were exposed to a diet containing 6.7% (v/v) ethanol or an isocaloric control diet during the last two weeks of gestation. At birth, the right infraorbital nerves of the pups were transected. The numbers of neurons in the principal sensory nucleus of the trigeminal nerve (PSN) on both sides of the pons were examined at various prenatal and early postnatal timepoints. The numbers of pyknotic and argyrophilic PSN cells were also counted. Ethanol delayed and reduced (19.9%) the prenatal acquisition of PSN neurons. The postnatal decline in neuronal number (indicative of neuronal death) was significantly increased (10.6%) by ethanol. Likewise, the numbers of pyknotic and silver-stained cells were significantly higher in ethanol-treated rats. Lesion of the infraorbital nerve induced significant transsynaptic neuronal death in the control rats. Ethanol increased the amount of death caused by the lesion;however, it altered neither the timing of the neuronal loss nor the incidence of pyknosis or silver-staining. Therefore, ethanol affects both neuronal acquisition and survival;the greater effect being on neuronal acquisition. The timing and morphology of dying cells indicate that regardless of the cause (natural processes, ethanol-induced, or lesion-induced), neurons die in the developing PSN by the same mode.

关键词：年龄因素轴突切断术细胞计数细胞死亡/药物作用中枢神经系统抑制药/药理学乙醇/药理学纵向研究上颌神经/细胞学上颌神经/生理学神经元传入/细胞学神经元传入/药物作用产前暴露迟发效应大鼠 Sprague-Dawley 三叉神经核/细胞学动物女(雌)性妊娠大鼠

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Genetic differences in alcohol sensitivity and the inheritance of alcoholism risk

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PSYCHOLOGICAL MEDICINE 1999年第5期29卷 1069-1081页

作者： Heath, AC Madden, PAF Bucholz, KK Dinwiddie, SH Slutske, WS Bierut, LJ Rohrbaugh, JW Statham, DJ Dunne, MP Whitfield, JB Martin, NG Washington Univ Sch Med Dept Psychiat St Louis MO 63110 USA Univ Missouri Dept Psychol Columbia MO USA Chicago Med Sch Dept Psychiat N Chicago IL USA Queensland Inst Med Res Dept Epidemiol & Populat Hlth Brisbane Qld 4006 Australia Queensland Univ Technol Dept Epidemiol Brisbane Qld Australia Royal Prince Alfred Hosp Dept Clin Biochem Sydney NSW Australia

Background. Substantial evidence exists for an important genetic contribution to alcohol dependence risk in women and men. It has been suggested that genetically determined differences in alcohol sensitivity may represent one pathway by which an increase in alcohol dependence risk occurs. Methods. Telephone interview follow-up data were obtained on twins from male, female and unlike-sex twin pairs who had participated in an alcohol challenge study in 1979-81, as well as other pairs from the same Australian twin panel surveyed by mail in 1980-82. Results. At follow-up, alcohol challenge men did not differ from other male twins from the same age cohort on measures of lifetime psychopathology or drinking habits;but alcohol challenge women were on average heavier drinkers than other women. A composite alcohol sensitivity measure, combining subjective intoxication and increase in body-sway after alcohol challenge in 1979-81, exhibited high heritability (60%). Parental alcoholism history was weakly associated with decreased alcohol sensitivity in women, but not after adjustment for baseline drinking history, or in men. High alcohol sensitivity in men was associated with substantially reduced alcohol dependence risk (OR = 0.05, 95% CI 0.01-0.39). Furthermore, significantly decreased (i.e;low) alcohol sensitivity was observed in non-alcoholic males whose MZ co-twin had a history of alcohol dependence, compared to other non-alcoholics. These associations remained significant in conservative analyses that controlled for respondents' alcohol consumption levels and alcohol problems in 1979-81. Conclusions. Men (but not women) at increased genetic risk of alcohol dependence (assessed by MZ co-twin's history of alcohol dependence) exhibited reduced alcohol sensitivity. Associations with parental alcoholism were inconsistent.

关键词：慢性酒精中毒/病因学慢性酒精中毒/遗传学中枢神经系统抑制药/药理学乙醇/药理学疾病遗传易感性危险性评估性别因素成年人女(雌)性人类男(雄)性中年人

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Inhibitory mechanism of costunolide, a sesquiterpene lactone isolated from Laurus nobilis, on blood-ethanol elevation in rats:: Involvement of inhibition of gastric emptying and increase in gastric juice secretion

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ALCOHOL AND ALCOHOLISM 2002年第2期37卷 121-127页

作者： Matsuda, H Shimoda, H Ninomiya, K Yoshikawa, M Kyoto Pharmaceut Univ Yamashina Ku Kyoto 6078412 Japan

Basic inhibitory mechanisms of costunolide and its active component, alpha-methylene-gamma-butyrolactone (alpha-MGBL), on blood-ethanol elevation were investigated in rats. In normal rats, blood-ethanol elevation (30 min later) induced by 20% (v/v) ethanol [5 ml/kg, per os (p.o.)] was strongly inhibited by pretreatment (30 min earlier) with costunolide and alpha-MGBL (50 mg/kg, p.o.). In pylorus-ligated rats given ethanol, blood-ethanol level (30 min) was barely elevated compared with that of normal rats, Neither costunolide nor alpha-MGBL affected the blood-ethanol elevation in pylorus-ligated rats or that induced by intraperitoneal and intraduodenal ethanol administration. Moreover, these compounds given orally induced no irreversible changes in alcohol dehydrogenase activity in rat liver. We continuously investigated the rate of gastric emptying in rats given various test meals. Costunolide and alpha-MGBL suppressed gastric emptying in rats given 20% ethanol and 1% sodium carboxymethyl cellulose, alpha-MGBL (50 mg/kg), but not costunolide, suppressed gastric emptying in 20% glucose-loaded rats. In an in vitro experiment, alpha-MGBL contracted the pylorus strip at a high concentration (20 mM), which was the estimated concentration in the stomach when the substance was given orally in vivo. These findings suggested that alpha-MGBL constricted the pylorus and caused delay of gastric emptying. Moreover, both compounds increased gastric fluid secretion with pepsin and mucus. In conclusion, the inhibitory effects of costunolide and alpha-MGBL on blood-ethanol elevation were based on inhibition of gastric emptying and dilution of the ethanol concentration by the increased gastric fluid.

关键词： 4-丁内酯/类似物和衍生物 4-丁内酯/化学 4-丁内酯/药理学投药口服醇脱氢酶/拮抗剂和抑制剂抗溃疡病药/化学抗溃疡病药/药理学中枢神经系统抑制药/拮抗剂和抑制剂中枢神经系统抑制药/血液中枢神经系统抑制药/药理学剂量效应关系药物酶抑制剂/化学酶抑制剂/分离和提纯酶抑制剂/药理学乙醇/拮抗剂和抑制剂乙醇/血液乙醇/药理学胃排空/药物作用胃排空/生理学胃液/药物作用胃液/代谢月桂/化学肌收缩/药物作用肌收缩/生理学幽门/药物作用幽门/生理学大鼠 Wistar 倍半萜类/化学倍半萜类/分离和提纯倍半萜类/药理学动物男(雄)性大鼠

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