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离体海马脑片CA1区锥体细胞缺氧期间电生理指标变化与n-甲基-d-天冬氨酸受体阻断剂MK801干预后的变化(英文)

中国临床康复 2005年第33期9卷 146-148页

作者：张育才曾因明王钧解放军第九七医院麻醉科江苏省徐州市221004 徐州医学院江苏省麻醉学重点实验室江苏省徐州市221002 Healthy-Science Center State University of New YorkNew YorkUSA

背景:海马CA1区锥体细胞是对缺血缺氧性损伤最为敏感的神经元,缺血缺氧后海马CA1区锥体细胞膜电位表现为缺氧早期细胞膜的超极化,随着缺氧时间的延长,细胞膜发生缓慢去极化及快速去极化,引起神经元不可逆性损伤。目的:应用细胞内记录技... 详细信息

背景:海马CA1区锥体细胞是对缺血缺氧性损伤最为敏感的神经元,缺血缺氧后海马CA1区锥体细胞膜电位表现为缺氧早期细胞膜的超极化,随着缺氧时间的延长,细胞膜发生缓慢去极化及快速去极化,引起神经元不可逆性损伤。目的:应用细胞内记录技术,观察n-甲基-d-天冬氨酸受体阻断剂MK801对离体海马脑片CA1区锥体细胞缺氧期间电生理指标的变化。设计:观察对比实验。单位:解放军第九七医院,徐州医学院江苏省麻醉学重点实验室,Healthy-ScienceCenter,StateUniversityofnewYork。材料:实验于2002-09/2003-02在美国纽约州立大学医学中心完成。选择成年雄性Sd大鼠5只,预吸纯氧3min后以体积分数为0.02的异氟醚麻醉,快速断头取脑,制备离体海马脑片。方法:大鼠海马脑片随机分为单纯缺氧组和MK801组,每组10个。单纯缺氧组海马脑片给予10min缺氧;而MK801组的海马脑片在缺氧前10min及10min的缺氧期间分别应用100μmol/L的MK801。所有脑片均给予60min的复氧。应用细胞内记录技术记录海马CA1区神经元缓慢去极化及快速去极化的时间、快速去极化的幅度。在复氧末,应用细胞内注入电流及经Schaffer通路刺激,观察神经元对刺激的反应。主要观察指标:①两组海马CA1区锥体神经元缓慢去极化速率。②两组海马CA1区神经元的快速去极化时间。③两组海马CA1区神经元的快速去极化幅度。④MK801对海马CA1区神经元功能恢复的影响。结果:①海马CA1区锥体神经元缓慢去极化速率:单纯缺氧组显著高于MK801组[(0.20±0.05)mV/s,(0.08±0.03)mV/s,(P<0.05)]。②海马CA1区神经元的快速去极化时间:MK801组显著高于单纯缺氧组[(537±139)s,(261±26)s,(P<0.05)]。③海马CA1区神经元的快速去极化的幅度:MK801组显著小于单纯缺氧组[(4±13)mV,(53±7)mV,(P<0.05)。④在复氧末期,MK801组的10个神经元中,9个神经元恢复对刺激的反应。结论:n-甲基-d-天冬氨酸受体阻断剂MK801可以显著降低缺氧引起的神经元缓慢去极化速率,延缓神经元快速去极化的发生及降低快速去极化的幅度,说明n-甲基-d-天冬氨酸受体阻断剂可显著减轻神经元的缺氧性损伤,促进复氧后神经元功能的恢复。

关键词：脑受体 n-甲基-d-天冬氨酸/拮抗剂和抑制剂细胞低氧海马电生理学

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Reduced ethanol inhibition of n-methyl-d-aspartate receptors by deletion of the nR1 C0 domain or overexpression of α-actinin-2 proteins

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JOURnAL OF BIOLOGICAL CHEMISTRY 2000年第20期275卷 15019-15024页

作者： Anders, dL Blevins, T Smothers, CT Woodward, JJ Virginia Commonwealth Univ Dept Pharmacol & Toxicol MCV Stn Richmond VA 23298 USA

The depressant actions of ethanol on central nervous system activity appear to be mediated by its actions on a number of important membrane associated ion channels including the n-methyl-d-aspartate (MMdA) subtype of ionotropic glutamate receptor. Although no specific site of action for ethanol on the nMdA receptor has been found, previous studies suggest that the ethanol sensitivity of the receptor may be affected by intracellular C-terminal domains of the receptor that regulate the calcium-dependent inactivation of the receptor. In the present study, co-expression of the nR2A subunit and an nR1 subunit that lacks the alternatively spliced intracellular C1 cassette did not reduce the effects of ethanol on channel function as measured by patch-clamp electrophysiology, Full inhibition was also observed in cells expressing an nR1 subunit truncated at the end of the CO domain (nR1(863stop)). However, the inhibitory effects of ethanol were reduced by expression of an nR1 CO domain deletion mutant (nR1 delta(839-863)), truncation mutant (nR1(858stop)), or a triple-point mutant (Arg to Ala, Lys to Ala, and Asn to Ala at 859-861) previously shown to significantly reduce calcium-dependent inactivation. A similar reduction in the effects of ethanol on wild-type nR1/2A but not nR1/2B or nR1/2C receptors was observed after co-expression of full-length or truncated human skeletal muscle alpha-actinin-2 proteins that produce a functional knockout of the C0 domain. The effects of ethanol on hippocampal and cortical nMdA-induced currents were similarly attenuated in low calcium recording conditions, suggesting that a C0domain-dependent process may confer additional ethanol sensitivity to nMdA receptors.

关键词：辅肌动蛋白/遗传学辅肌动蛋白/代谢选择性剪接氨基酸序列氨基酸取代钙/药理学细胞系细胞培养的乙醇/药理学海马/生理学分子序列数据诱变定点锥体细胞/药物作用锥体细胞/生理学受体 n-甲基-d-天冬氨酸/拮抗剂和抑制剂受体 n-甲基-d-天冬氨酸/化学受体 n-甲基-d-天冬氨酸/遗传学重组蛋白质类/拮抗剂和抑制剂重组蛋白质类/化学序列缺失转染动物人类大鼠

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Activity of putative cognition enhancers in kynurenate test performed with human neocortex slices

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JOURnAL OF PHARMACOLOGY And EXPERIMEnTAL THERAPEUTICS 1999年第1期290卷 423-428页

作者： Pittaluga, A Pattarini, R Andrioli, GC Viola, C Munari, C Raiteri, M Univ Genoa Dept Expt Med Pharmacol & Toxicol Sect I-16148 Genoa Italy Univ Genoa Osped Galliera Div Neurochirurg I-16148 Genoa Italy Univ Genoa Clin Neurochirurg I-16148 Genoa Italy

Some cognition enhancers were previously shown to potently prevent antagonism of the n-methyl-d-aspartate (nMdA)evoked release of norepinephrine (nE) brought about in slices of rat hippocampus by kynurenic acid, an endogenous nMdA receptor blocker. We have examined the impact of putative nootropic agents in the kynurenate test performed with slices of human cerebral cortex from patients undergoing neurosurgery. in slices of human neocortex, local application of nMdA evoked release of [(3)H]nE;the effect of nMdA was antagonized by several nMdA receptor antagonists, including kynurenic acid. The antagonism of the nMdA-evoked [(3)H]nE release produced by 300 mu M kynurenate was potently (EC(50) <10 mu M) prevented by most of the nootropics tested, including aniracetam, oxiracetam, d-cycloserine, and the glutamate analog CR 2249 (but not its enantiomer CR 2361). nicotine or tacrine (up to 10 mu M) did not show any effect in the kynurenate test. nicotine (30-100 mu M) itself increased the release of [(3)H]nE;interestingly, the nicotine-evoked overflow was blocked not only by the nicotin receptor antagonist mecamylamine but also by nMdA receptor antagonists, suggesting an indirect mechanism mediated by glutamate/aspartate release. To conclude, the similarities between the data obtained here with human neocortex slices and those previously obtained in the rat indicate that the kynurenate test performed with rat brain slices may represent a useful biochemical assay to study cognition-enhancing drugs.

关键词：兴奋性氨基酸激动剂/药理学兴奋性氨基酸拮抗剂/药理学犬尿烯酸/拮抗剂和抑制剂犬尿烯酸/药理学 n-甲基天冬氨酸/药理学新皮质/药物作用新皮质/代谢脑代谢改善药/药理学去甲肾上腺素/代谢受体 n-甲基-d-天冬氨酸/拮抗剂和抑制剂人类

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Seizure-like events in disinhibited ventral slices of adult rat hippocampus

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JOURnAL OF nEUROPHYSIOLOGY 1999年第5期82卷 2130-2142页

作者： Borck, C Jefferys, JGR Univ Birmingham Sch Med Dept Neurophysiol Div Neurosci Birmingham B15 2TT W Midlands England Univ London Imperial Coll Sci Technol & Med Dept Physiol & Biophys St Marys Hosp Sch Med London W2 1PG England

Epileptic discharges lasting 2-90 s, were studied in vitro in slices from the ventral hippocampus of adult rats, in which inhibition was blocked acutely with bicuculline methiodide (BMI, 5-30 mu M) and potassium ([K+](o)) raised to 5 mM. These seizure-like events (SLEs) comprised three distinct phases, called here primary, secondary, and tertiary bursts. Primary bursts lasted 90-150 ms. Secondary bursts lasted a further 70-250 ms, comprising a short series of afterdischarges riding on the same depolarization as the primary burst. Finally a train of tertiary bursts started with a peak frequency of 5-10 Hz and could last >1 min. Slices from the ventral hippocampus showed significantly higher susceptibility to SLEs than did dorsal slices. SLEs proved sensitive to alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor antagonists. They were insensitive to n-methyl-d-aspartate (nMdA) receptor antagonists;50 mu l d-2-amino-5-phosphonopentanoic acid (d-AP5) did block the transient secondary bursts selectively. SLEs were restricted to the hippocampus proper even if the entorhinal cortex was present. Entorhinal bursts could last <2 s and were only coupled with hippocampal bursts in a minority of slices. Reentry of epileptic bursts occasionally occurred during interictal discharges, but not during the later stages of SLEs. Full-length SLEs always started in CA3 region and could be recorded in minislices containing CA3 plus dentate hilus. Ion-sensitive microelectrodes revealed that interictal discharges were followed by short (2-3 s) [K+](o) waves, peaking at similar to 7.5 mM. SLEs were always accompanied by increases in [K+](o) reaching similar to 8.5 mM at the start of tertiary bursts;[K+](o) then increased more slowly to a ceiling of 11-12 mM. After the end of each SLE, [K+](o) fell back to baseline within 10-15 s. SLEs were accompanied by significant increase in synaptic activity, compared with baseline and/or interictal activity, estimated by the varian

关键词： 2-氨基-5-膦酰基戊酸盐/药理学荷包牡丹碱/类似物和衍生物荷包牡丹碱/药理学电刺激癫痫/病理生理学诱发电位/药物作用诱发电位/生理学兴奋性氨基酸拮抗剂/药理学海马/药物作用海马/生理学海马/病理生理学氯化钾/药理学锥体细胞/药物作用锥体细胞/生理学大鼠 Sprague-dawley 受体 AMPA/拮抗剂和抑制剂受体 n-甲基-d-天冬氨酸/拮抗剂和抑制剂发作/病理生理学动物男(雄)性大鼠

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Quantitative analysis of tetrapentylammonium-induced blockade of open n-methyl-d-aspartate channels

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BIOPHYSICAL JOURnAL 2000年第3期79卷 1324-1335页

作者： Sobolevsky, AI Russian Acad Med Sci Inst Gen Pathol & Pathol Physiol Moscow 125315 Russia

The blockade of open n-methyl-d-aspartate (nMdA) channels by tetrapentylammonium (TPentA) in acutely isolated rat hippocampal neurons was studied using whole-cell patch-clamp techniques. TPentA prevented the closure of the nMdA channel following what is known as the foot-in-the-door mechanism. Hooked tail currents appearing after termination of the agonist (aspartate) and TPentA coapplication were analyzed quantitatively according to the corresponding sequential kinetic model. Studies of the hooked tail current amplitude and the degree of the stationary current inhibition dependence on the blocker concentration led to a new method for estimation of fast foot-in-the-door blocker binding/unbinding rate constants. The application of this method to the nMdA channel blockade by TPentA allowed finding the values of its binding (1.48 mu M-1 s(-1)) and unbinding (14 s(-1)) rate constants. An analysis of the dependence of the electric charge carried during the hooked tail current on the blocker concentration led to a new method for estimation of the maximum nMdA channel open probability, P-o. The value of P-o found in experiments with TPentA was 0.04.

关键词：海马/生理学动力学膜电位/药物作用膜电位/生理学模型理论膜片钳术锥体细胞/药物作用锥体细胞/生理学季胺类化合物/药理学受体 n-甲基-d-天冬氨酸/拮抗剂和抑制剂动物大鼠

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Structure-activity relationship for a series of 2-substituted 1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indoles:: Potent subtype-selective inhibitors of n-methyl-d-aspartate (nMdA) receptors

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BIOORGAnIC & MEdICInAL CHEMISTRY LETTERS 1999年第11期9卷 1619-1624页

作者： Tamiz, AP Whittemore, ER Woodward, RM Upasani, RB Keana, JFW CoCensys Inc Irvine CA 92618 USA Univ Oregon Dept Chem Eugene OR 97403 USA

series of 2-substituted 1,2,3,4-tetrahydro-9H-pyrido[3,4-b] indoles was synthesized as potential antagonists for the nR1A/2B subtype of n-methyl-d-aspartate (nMdA) receptors. Assayed by electrical recording under steady-state conditions, 7-hydroxy-2-(4-phenylbutyl)-1,2,3,4-tetrahydropyrido-[3,4-b]indole (30) was the most potent compound in the series having an IC50 value of 50 nM at the MR1A/2B receptors. (C) 1999 Elsevier Science Ltd. All rights reserved.

关键词：吲哚类/化学合成吲哚类/化学吲哚类/药理学半数抑制浓度模型化学吡啶类/化学合成吡啶类/药理学受体 n-甲基-d-天冬氨酸/拮抗剂和抑制剂构效关系爪蟾属/胚胎学动物

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Preincisional dextromethorphan treatment for postoperative pain management after upper abdominal surgery

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WORLd JOURnAL OF SURGERY 2000年第5期24卷 512-517页

作者： Wu, CT Yu, JC Liu, ST Yeh, CC Li, CY Wong, CS Tri Serv Gen Hosp Dept Anesthesiol Taipei 100 Taiwan Natl Def Med Ctr Taipei 100 Taiwan Tri Serv Gen Hosp Dept Surg Div Gen Surg Taipei 100 Taiwan

Previous studies showed that ketamine, an n-methyl-d-aspartate (nMdA) receptor antagonist, provides a preemptive analgesic effect and preemptive analgesia improves postoperative pain management. The aim of this study was to examine whether premedication with destromethorphan (dM) improves postoperative pain management after upper abdominal surgery. Sixty (American Society of Anesthesiologists class 1 and 2 of either gender) patients scheduled for upper abdominal surgery were included in the study. Patients were randomly assigned to one of four groups: control, dM-10, dM-20, and dM-40. In the control group, chlorpheniramine maleate (CPM, 20 mg) was injected immediately before induction of anesthesia intramuscularly (IM). In the dM-IO, dM-20, and dM-40 groups, patients were premedicated with dM 10 mg, 20 mg, and 10 mg IM, respectively. After operation, patient-controlled analgesia (PCA) with morphine was given for pain relief. The time to the first PCA trigger, morphine consumption, pain scores, and analgesic-related side effects were recorded at 1, 2, 4, 24, 48, and 72 hours after surgery. The time to first PCA trigger for the control group was 17.8 +/- 1.4 minutes, for group dM-10 20.2 +/- 1.6 minutes, for group dM-20 32.4 +/- 1.9 minutes, and for dM-40 77.9 +/- 6.5 minutes. The morphine delivered and PCA triggering frequency were 5.5 +/- 0.5/11.3 +- 0.8 times for the controls, 5.5 +/- 0.4/ 14.1 +/- 1.3 times for dM-10, 3.1 +/- 0.3/6.3 +/- 1.2 times for dM-20, and 0.2 +/- 0.1/0.3 +/- 0.2 times for dM-40 during the first hour after operation. For the first day, the figures are 19.9 +/- 1.2/23.9 +/- 1.4 for the controls, 15.6 +/- 1.2/17.3 +/- 2.4 for dM-10, 12.6 +/- 0.7/15.9 +/- 1.6 for dM-20, and 5.0 +/- 0.21/5.6 +/- 0.9 for dM-40. On the first day, the cough pain stores were 6.67 +/- 0.23. 6.53 +/- 0.16, 6.67 +/- 0.23, and 5.73 +/- 0.18 for the controls, dM-10, dM-20, and dM-40 groups, respectively. All data showed dose-dependent better pain relief in dM-premedicate

关键词：腹部/外科学右美沙芬/治疗应用疼痛手术后/预防和控制手术前护理/方法受体 n-甲基-d-天冬氨酸/拮抗剂和抑制剂女(雌)性人类男(雄)性中年人

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Actions of the anticonvulsant remacemide metabolite AR-R12495AA on afferent-evoked spinal synaptic transmission in vitro and on models of acute and chronic inflammation in the rat

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JOURnAL OF PHARMACOLOGY And EXPERIMEnTAL THERAPEUTICS 2000年第3期294卷 876-883页

作者： Asghar, AUR Hasan, SS King, AE Univ Leeds Sch Biomed Sci Leeds LS2 9NG W Yorkshire England

The effects of the anticonvulsant remacemide [(+/-)-2-amino-n-(1-methyl-1,2-diphenylethyl)-acetamide hydrochloride] and its metabolite AR-R12495AA [(+/-)-1-methyl-1,2-diphenylelhylamine-monohydrochloride] on primary afferent synaptic transmission were assessed in the young rat spinal cord in vitro. Stimulation of dorsal roots at A- and C-afferent intensity elicited a dorsal root-evoked ventral root potential (dR-VRP) with a slowly decaying phase. Repetitive stimuli (2 Hz) produced summation of slow potentials and a cumulative ventral root depolarization (CVRd), a form of wind-up. Remacemide and ARR12495AA antagonized the dR-VRP slow peak t(1/2) decay and slow phase total duration at drug concentration of greater than or equal to 25 mu M. AR-R12495AA was approximately 2-fold more potent than remacemide. The most potent action was against the slow phase duration with IC50 values of 157 and 60 mu M for remacemide and AR-R12495A4, respectively. Both drugs at concentrations of greater than or equal to 100 mu M attenuated the dR-VRP fast peak amplitude (IC50 = 253 and 142 mu M, respectively). The amplitude of CVRd was reduced by remacemide and AR-R12495AA (IC50 = 195 and 111 mu M, respectively). MK-801 reduced dR-VRP fast peak amplitude (IC50 = 58 mu M), slow peak t(1/2) decay (IC50 = 60 mu M), slow phase duration (IC50 = 50 mu M), and CVRd amplitude (IC50 = 91 mu M). In behavioral studies, AR-R12495AA (i.p,) reduced the mechanical hyperalgesia and paw swelling that followed hind paw injection of carrageenan or Freund's complete adjuvant. These electrophysiological and behavioral data indicate further studies should be conducted on the efficacy of remacemide and AR-R12495AA as putative analgesics.

关键词：乙酰胺类/药理学急性病传入通路/生理学动物新生抗惊厥药/药理学慢性病地卓西平马来酸盐/药理学电刺激电生理学兴奋性氨基酸拮抗剂/药理学痛觉过敏/化学诱导痛觉过敏/药物疗法炎症/病理生理学苯乙胺类/药理学大鼠 Wistar 受体 n-甲基-d-天冬氨酸/拮抗剂和抑制剂脊髓/药物作用脊髓/生理学脊神经根/生理学突触传递/药物作用动物大鼠

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An oral vaccine against nMdAR1 with efficacy in experimental stroke and epilepsy

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SCIEnCE 2000年第5457期287卷 1453-1460页

作者： during, MJ Symes, CW Lawlor, PA Lin, J dunning, J Fitzsimons, HL Poulsen, d Leone, P Xu, RA dicker, BL Lipski, J Young, d Univ Auckland Sch Med Dept Mol Med Auckland New Zealand Univ Auckland Sch Med Dept Physiol Auckland New Zealand Thomas Jefferson Univ Philadelphia PA 19107 USA Jefferson Med Coll Dept Neurosurg Philadelphia PA 19107 USA

The brain is generally considered immunoprivileged, although increasing examples of immunological responses to brain antigens, neuronal expression of major histocompatibility class I genes, and neurological autoimmunity have been recognized. An adeno-associated virus (AAV) vaccine generated autoantibodies that targeted a specific brain protein, the nR1 subunit of the n-methyl-d-aspartate (nMdA) receptor. After peroral administration of the AAV vaccine, transgene expression persisted for at least 5 months and was associated with a robust humoral response in the absence of a significant cell-mediated response. This single-dose vaccine was associated with strong anti-epileptic and neuroprotective activity in rats for both a kainate-induced seizure model and also a middle cerebral artery occlusion stroke model at 1 to 5 months following vaccination. Thus, a vaccination strategy targeting brain proteins is feasible and may have therapeutic potential for neurological disorders.

关键词：投药口服抗体亲和力自身抗体/分析自身抗体/血液自身抗体/脑脊髓液自身抗体/免疫学血脑屏障依赖病毒/遗传学癫痫颞叶/病理学癫痫颞叶/治疗表位作图表位遗传载体海马/病理学肠黏膜/免疫学肠黏膜/代谢淋巴细胞活化运动活动受体 n-甲基-d-天冬氨酸/拮抗剂和抑制剂受体 n-甲基-d-天冬氨酸/生物合成受体 n-甲基-d-天冬氨酸/遗传学受体 n-甲基-d-天冬氨酸/免疫学癫痫持续状态/预防和控制卒中/治疗转基因接种疫苗 dnA/治疗应用动物大鼠

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Eosine-induced blockade of n-methyl-d-aspartate channels in acutely isolated rat hippocampal neurons

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MOLECULAR PHARMACOLOGY 2000年第2期57卷 334-341页

作者： Sobolevsky, AI Yelshansky, MV Khodorov, BI Inst Gen Pathol & Pathophysiol Moscow 125315 Russia

Acutely isolated rat hippocampal neurons were voltage-clamped in the whole-cell configuration. The currents through n-methyl-d-aspartate (nMdA) channels were elicited by fast application of aspartate in a Mg2+-free 3 mu M glycine-containing solution. Eosine, known as a potent reversible inhibitor of the plasma membrane Ca2+ pump, proved to be able to induce a blockade of nMdA channels. The eosine-induced inhibition of nMdA-mediated currents enhanced with eosine concentration (IC50 = 248 mu M) but did not depend on the membrane potential, agonist (aspartate) or coagonist (glycine) concentrations, pH, or the presence of spermine, ethanol, and the disulfide-reducing agents dithiothreitol and glutathione. Zn2+ inhibited nMdA channels with equal efficiency both in the presence and absence of eosine. These results suggest that eosine interacts with a new, previously unknown nMdA receptor regulatory site.

关键词：曙红黄-(YS)/药理学海马/药物作用海马/代谢神经元/药物作用神经元/代谢受体 n-甲基-d-天冬氨酸/拮抗剂和抑制剂动物大鼠

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