越鞠甘麦大枣汤增强突触可塑性产生抗抑郁样作用＊

目的 研究越鞠甘麦大枣汤的抗抑郁样作用并分析其对小鼠海马突触可塑性的影响。方法 昆明小鼠随机分为对照组和越鞠甘麦大枣汤组，给药24 h和7 天后进行悬尾测试（Tail suspension test，TST）和强迫游泳测试（Forced swimming test，FST）。运用电生理（electrophysiological experiment）技术检测小鼠海马区Schaffe侧枝-CA1的长时程增强（long term potentiation，LTP），利用western blot方法分析海马脑区α-氨基-3-羟基-5-甲基-4-异恶唑丙酸受体（α-amino-3-hydroxy-5-methyl-4-isoxazole-propionicacid receptor，AMPAR）和N-甲基-d-天冬氨酸受体（N-methyl-D-aspartate receptor， NMDAR）相关突触蛋白的表达水平。结果 给药24 h和7天后，与对照组小鼠相比，越鞠甘麦大枣汤给药组小鼠在TST（P <0.001）和FST（P <0.01）中的不动时间均明显降低，氯胺酮在给药24 h后显示出抗抑郁作用，但到第7天不能降低小鼠不动时间，越鞠甘麦大枣汤显示出更持久的抗抑郁样作用。电生理实验中，越鞠甘麦大枣汤可增强小鼠海马区的LTP（P < 0.001）。Western blot结果显示，GluR1、NR2B以及NR1的表达水平在给药后均明显增加（P < 0.05）。结论 越鞠甘麦大枣汤可能通过增强昆明小鼠海马脑区LTP，以及增加AMPA和NMDA受体相关突触蛋白的表达水平以提高突触传递效能，从而产生抗抑郁样作用。     

Movement disorders in children with anti‐NMDAR encephalitis and other autoimmune encephalopathies

Accurate recognition of movement disorder phenomenology may differentiate children with anti‐N‐methyl D‐aspartate receptor (NMDAR) encephalitis, autoimmune basal ganglia encephalitis (BGE), and Sydenham's chorea (SC). Three neurologists blinded to the diagnoses recorded dominant and associated movement disorders seen on videos of 31 patients with anti‐NMDAR encephalitis (n = 10), BGE (n = 12), and SC (n = 9). Stereotypy was only seen in anti‐NMDAR encephalitis (8/10) and not in BGE and SC (P < 0.001). Perseveration was only seen in anti‐NMDAR encephalitis (5/10) and not in BGE and SC (P < 0.001). Akinesia was more commonly seen in BGE (5/12) than in anti‐NMDAR encephalitis (1/10, P = 0.097). Tremor was more commonly seen in BGE (5/12) than in anti‐NMDAR encephalitis (1/10, P = 0.097). Chorea was seen in all groups: anti‐NMDAR encephalitis (4/10), BGE (3/12), and SC (9/9). Likewise, dystonia was seen in all groups: anti‐NMDAR encephalitis (6/10), BGE (7/12), and SC (2/9). Stereotypies or perseveration are suggestive of anti‐NMDAR encephalitis, whereas their absence and the presence of akinesia and tremor is more suggestive of BGE. Chorea and dystonia are least discriminating. © 2014 International Parkinson and Movement Disorder Society     

白香丹胶囊对PMDD肝气逆证模型大鼠海马NMDAR1亚基蛋白分布及表达的影响

目的：以PMDD肝气逆证模型大鼠为载体,研究PMDD肝气逆证的病机以及白香丹胶囊对本病的干预机制。方法：改进情志刺激为主多因素分段刺激造模法,复制PMDD肝气逆证大鼠模型;用白香丹胶囊和氟西汀分散片予以干预,最后进行旷场实验评价模型;免疫荧光技术和蛋白印迹技术检测大鼠海马CA1、CA3区NMDAR1受体亚基分布情况及表达水平。结果：模型组大鼠与正常组相比较,其旷场实验的运动总距离明显增加（P<0.001）、中央区域的进入次数及停留在中央区域的时间显著减少（P<0.01）,海马CA1、CA3区细胞呈现稀疏杂乱的分布状态,且同氟西汀组一样,其NMDAR1蛋白表达量明显降低;白香丹组大鼠与模型组相比较,其旷场实验的运动总距离明显降低（P<0.05）、中央区域的进入次数和停留在中央区域的时间明显增多（P<0.05）,而氟西汀组与之相比,除了运动总距离明显降低（P<0.05）,其它指标不存在显著性差异,同氟西汀组一样,白香丹组大鼠海马CA1、CA3区细胞分布、排列无显著异常,但其NMDAR1蛋白表达量显著升高（P<0.0001）;氟西汀组大鼠与白香丹组相比较,其NMDAR1蛋白表达量明显降低（P<0.05）。结论：情志刺激为主多因素分段刺激造模法对大鼠学习记忆能力的损伤机制可能与大鼠海马CA1、CA3区神经细胞的减少和NMDAR1亚基的表达量降低有关,白香丹胶囊通过调整NMDAR1蛋白表达量纠正上述异常改变,从而改善大鼠的学习记忆能力。     

Expression pattern of NMDA receptors reveals antiepileptic potential of apigenin 8-C-glucoside and chlorogenic acid in pilocarpine induced epileptic mice

The present study was aimed to evaluate the effect of apigenin 8-C-glucoside (Vitexin) and chlorogenic acid on epileptic mice induced by pilocarpine and explored its possible mechanisms. Intraperitonial administration of pilocarpine (85 mg/kg) induced seizure in mice was assessed by behavior observations, which is significantly (p > 0.05) reduced by apigenin 8-C-glucoside (AP8CG) (10 mg/kg) and chlorogenic acid (CA) (5 mg/kg), similar to diazepam. Seizure was accompanied by an imbalance in the levels of Gamma-aminobutyric acid (GABA) and glutamate in the pilocarpine administered group. Moreover, convulsion along with reduced acetylcholinesterase, increased monoamine oxidase and oxidative stress was observed in epileptic mice brain. AP8CG and CA significantly restored back to normal levels even at lower doses. Further, increased lipid peroxidation and nitrite content was also significantly attenuated by AP8CG and CA. However, CA was found to be more effective when compared to AP8CG. In addition, the mRNA expression of N-methyl-d-aspartate receptor (NMDAR), mGluR1 and mGlu5 was significantly (P ≤ 0.05) inhibited by AP8CG and CA in a lower dose. The mRNA expression of GRIK1 did not differ significantly in any of the group and showed a similar pattern of expression. Our result shows that AP8CG and CA selectively inhibit NMDAR, mGluR1 and mGlu5 expression. Modification in the provoked NMDAR calcium response coupled with neuronal death. Hence, these findings underline that the polyphenolics, AP8CG and CA have exerted antiepileptic and neuroprotective activity by suppressing glutamate receptors.     

右归饮对去卵巢血管钙化大鼠NMDAR1蛋白表达的影响

目的:观察右归饮对去卵巢血管钙化中谷氨酸受体变化的影响。方法:将40只远交群雌性大鼠(sprague dawley,SD)随机分为假手术组、模型组、低剂量组、高剂量组和辛伐他汀组,采用皮下注射华法令加维生素D3制备血管钙化模型,造模后,低剂量组、高剂量组分别给予右归饮浓缩液5.35 g/(kg·d)和16.05 g/(kg·d)灌胃,辛伐他汀组给予辛伐他汀混悬水溶液5.25 mg/(kg·d)灌胃,假手术组和模型组给予等量生理盐水,共12周;实验结束时采集血浆标本测定雌二醇及血脂四项,采用苏木精-伊红染色法(HE染色)观察动脉钙化的情况,并采用免疫印迹试验检测N-甲基-D-天门冬氨酸受体(NMDAR,NR)1蛋白的表达。结果 :模型组大鼠雌二醇、甘油三酯含量和NR1蛋白表达降低,P<0.05,低密度脂蛋白升高,P<0.05;高剂量组和低剂量组雌二醇、甘油三酯含量升高,NR1蛋白表达增加,P<0.05;高剂量组、低剂量组和辛伐他汀组低密度脂蛋白的含量降低,P<0.05;辛伐他汀组雌二醇无变化,P>0.05。结论:右归饮可抑制大鼠去卵巢血管钙化的形成,可能与提高雌激素水平和NR1蛋白的表达有关。     

Enriched environment has limited capacity for the correction of hippocampal memory-dependent schizoid behaviors in rats with early postnatal NMDAR dysfunction

Pre- and early postnatal stress can cause dysfunction of the N-methyl-d-aspartate receptor (NMDAR) and thereby promote the development of hippocampus memory-dependent schizoid abnormalities of navigation in space, time, and knowledge. An enriched environment improves mental abilities in humans and animals. Whether an enriched environment can prevent the development of schizoid symptoms induced by neonatal NMDAR dysfunction was the central question of our paper. The experimental animals were Wistar rats. Early postnatal NMDAR dysfunction was created by systemic treatment of rat pups with the NMDAR antagonist MK-801 at PD10–20 days. During the development period (PD21–90 days), the rats were reared in cognitively and physically enriched cages. Adult age rats were tested on navigation based on pattern separation and episodic memory in the open field and on auto-hetero-associations based on episodic and semantic memory in a step-through passive avoidance task. The results showed that postnatal NMDAR antagonism caused abnormal behaviors in both tests. An enriched environment prevented deficits in the development of navigation in space based on pattern separation and hetero-associations based on semantic memory. However, an enriched environment was unable to rescue navigation in space and auto-associations based on episodic memory. These data may contribute to the understanding that an enriched environment has a limited capacity for therapeutic interventions in protecting the development of schizoid syndromes in children and adolescents.     

Targeting neuronal nitric oxide synthase as a valuable strategy for the therapy of neurological disorders

The management of neurological disorders have huge and increasing human and economic costs. De-spite this, there is a scarcity of effective therapeutics, and there is an extreme urgency for new and real treatments. In this short review we analyze some promising advancements in the search of new bioactive molecules targeting neuronal nitric oxide synthase (nNOS), an enzyme deputed to the biosynthesis of nitric oxide (NO). In different conditions of neuronal damages, this molecule is overproduced, contribut-ing to the pathogenesis and progression of neuronal diseases. Two main approaches to modulate nNOS are discussed: a ifrst one consisting in the direct inhibition of the enzyme by means of small organic molecules, which can be also active against other different targets involved in such diseases. A second section is dedi-cated to molecules able to prevent the formation of the ternary complex N-methyl-D-aspartate (NMDA)-type glutamate receptors, postsynaptic density-95 (PSD95) protein-nNOS, which is necessary to activate the latter for the biosynthesis of NO.     

Effects of Prenatal Ethanol Exposure on Brain Region NMDA-Mediated Increase in Intracellular Calcium and the NMDAR1 Subunit in Forebrain

The effects of prenatal ethanol exposure on N -methyl-d-aspartate (NMDA)-mediated calcium entry into neonatal dissociated neurons from hippocampus, forebrain, and cerebellum were investigated. Dissociated cells were isolated from less than 1-day-old pups of prenatally exposed, pair-fed control and ad libitum control groups and loaded with fura-2. Prenatal ethanol exposure significantly reduced the NMDA-stimulated increase in intracellular calcium in all three brain regions compared to the two control groups. These findings are very similar to those previously observed in neonatal dissociated whole brain neurons using the same ethanol exposure protocol. Studies were also conducted using forebrain to determine if prenatal ethanol exposure alters NMDAR1 subunit protein expression in this major brain area; however, the results indicated no significant differences between ethanol-exposed and control groups.     

Novel nootropic drug sunifiram enhances hippocampal synaptic efficacy via glycine‐binding site of N‐methyl‐D‐aspartate receptor

Sunifiram is a novel pyrrolidone nootropic drug structurally related to piracetam, which was developed for neurodegenerative disorder like Alzheimer's disease. Sunifiram is known to enhance cognitive function in some behavioral experiments such as Morris water maze task. To address question whether sunifiram affects N‐methyl‐D ‐aspartate receptor (NMDAR)‐dependent synaptic function in the hippocampal CA1 region, we assessed the effects of sunifiram on NMDAR‐dependent long‐term potentiation (LTP) by electrophysiology and on phosphorylation of synaptic proteins by immunoblotting analysis. In mouse hippocampal slices, sunifiram at 10–100 nM significantly enhanced LTP in a bell‐shaped dose‐response relationship which peaked at 10 nM. The enhancement of LTP by sunifiram treatment was inhibited by 7‐chloro‐kynurenic acid (7‐ClKN), an antagonist for glycine‐binding site of NMDAR, but not by ifenprodil, an inhibitor for polyamine site of NMDAR. The enhancement of LTP by sunifilam was associated with an increase in phosphorylation of α‐amino‐3‐hydroxy‐5‐methylisozazole‐4‐propionate receptor (AMPAR) through activation of calcium/calmodulin‐dependent protein kinase II (CaMKII) and an increase in phosphorylation of NMDAR through activation of protein kinase Cα (PKCα). Sunifiram treatments at 1–1000 nM increased the slope of field excitatory postsynaptic potentials (fEPSPs) in a dose‐dependent manner. The enhancement was associated with an increase in phosphorylation of AMPAR receptor through activation of CaMKII. Interestingly, under the basal condition, sunifiram treatments increased PKCα (Ser‐657) and Src family (Tyr‐416) activities with the same bell‐shaped dose‐response curve as that of LTP peaking at 10 nM. The increase in phosphorylation of PKCα (Ser‐657) and Src (Tyr‐416) induced by sunifiram was inhibited by 7‐ClKN treatment. The LTP enhancement by sunifiram was significantly inhibited by PP2, a Src family inhibitor. Finally, when pretreated with a high concentration of glycine (300 μM), sunifiram treatments failed to potentiate LTP in the CA1 region. Taken together, sunifiram stimulates the glycine‐binding site of NMDAR with concomitant PKCα activation through Src kinase. Enhancement of PKCα activity triggers to potentiate hippocampal LTP through CaMKII activation. © 2013 Wiley Periodicals, Inc.