小鼠逆行性遗忘动物模型的建立

目的 探讨用电击、缺氧、麻醉等处理方法 建立小鼠逆行性遗忘动物模型的可行性及优劣.方法 将72只昆明小鼠分为对照组及电休克、缺氧、丙泊酚、电休克+缺氧、电休克+丙泊酚5个处理组.先给予各组相同的避暗训练以建立避暗行为,随后分别给予各处理组120～180 V电击、密闭容器内缺氧、腹腔注射0.3 mL丙泊酚、120～180 V电击+密闭容器内缺氧、120～180 V电击+腹腔注射0.3 mL丙泊酚相应处理.次日开始用暗箱观察各组小鼠的步入潜伏期,以分析避暗行为的变化.结果对照组小鼠在避暗训练后24 h(第4天)的步入潜伏期为(111.7±17.2)S,缺氧组、电休克+缺氧组、电休克+丙泊汾组与对照组相比差异均有统计学意义(P<0.05);电休克组、缺氧组、电休克+缺氧组、电休克+丙泊酚组4组均有部分小鼠步入潜伏期明显缩短至30 s以内,其发生率分别为43.8%、45.4%、66.7%、60%,而丙泊酚组步入潜伏期无明显变化.第5天、第8天观察显示,步入潜伏期缩短的小鼠中个别出现恢复.结论 电休克、缺氧、电休克+缺氧、电休克+丙泊酚处理后的小鼠中部分可出现逆行性遗忘表现,以电休克+缺氧组建模的成功率最高:已出现逆行性遗忘的小鼠中部分可在后期恢复;单纯丙泊酚不能引起逆行性遗忘.     

小鼠逆行性遗忘动物模型的建立

目的 探讨用电击、缺氧、麻醉等处理方法 建立小鼠逆行性遗忘动物模型的可行性及优劣.方法 将72只昆明小鼠分为对照组及电休克、缺氧、丙泊酚、电休克+缺氧、电休克+丙泊酚5个处理组.先给予各组相同的避暗训练以建立避暗行为,随后分别给予各处理组120～180 V电击、密闭容器内缺氧、腹腔注射0.3 mL丙泊酚、120～180 V电击+密闭容器内缺氧、120～180 V电击+腹腔注射0.3 mL丙泊酚相应处理.次日开始用暗箱观察各组小鼠的步入潜伏期,以分析避暗行为的变化.结果对照组小鼠在避暗训练后24 h(第4天)的步入潜伏期为(111.7±17.2)S,缺氧组、电休克+缺氧组、电休克+丙泊汾组与对照组相比差异均有统计学意义(P<0.05);电休克组、缺氧组、电休克+缺氧组、电休克+丙泊酚组4组均有部分小鼠步入潜伏期明显缩短至30 s以内,其发生率分别为43.8%、45.4%、66.7%、60%,而丙泊酚组步入潜伏期无明显变化.第5天、第8天观察显示,步入潜伏期缩短的小鼠中个别出现恢复.结论 电休克、缺氧、电休克+缺氧、电休克+丙泊酚处理后的小鼠中部分可出现逆行性遗忘表现,以电休克+缺氧组建模的成功率最高:已出现逆行性遗忘的小鼠中部分可在后期恢复;单纯丙泊酚不能引起逆行性遗忘.     

γ-氨基丁酸能神经元在丙泊酚缓解抑郁模型大鼠电休克后脑损伤中的作用

目的探讨γ-氨基丁酸(γ-aminobutyric acid,GABA)能神经元在丙泊酚缓解抑郁大鼠电休克(electroconvulsive shock,ECS)后学习记忆损伤中的作用。方法将慢性温和不可预见性应激(chronic unpredictable mild stress,CUMS)法建模成功的36只抑郁症模型大鼠随机分为丙泊酚+电休克组、电休克组和抑郁组,另设同批次未建模的12只健康大鼠为对照组。丙泊酚+电休克组用丙泊酚联合电休克治疗,电休克组行电休克治疗,抑郁组与对照组行伪电休克处理。治疗完毕行Morris水迷宫实验评估大鼠空间学习记忆能力;ELISA法检测海马GABA浓度;免疫组化法和Western-blot检测海马GABAARα5的蛋白表达。结果Morris水迷宫实验结果提示,电休克组逃避潜伏期最长,空间探索时间最短(P<0.05);电休克组和丙泊酚+电休克组比抑郁组逃避潜伏期延长,空间探索时间缩短(P<0.05);与对照组比较,其余各组逃避潜伏期缩短,空间探索时间延长(P<0.05)。蛋白表达方面,与对照组相比,抑郁组GABA含量下降,GABAARα5表达水平降低(P<0.05),电休克组和丙泊酚+电休克组GABA含量和GABAARα5蛋白表达升高(P<0.05);与电休克组相比,丙泊酚+电休克组GABA含量下降,GABAARα5蛋白表达升高(P<0.05)。结论丙泊酚在电休克过程中具有脑保护作用,其机制可能与上调海马GABA能神经系统相关递质和受体的表达有关。     

逍遥散对慢性应激下拟阿尔茨海默病小鼠行为学和自由基代谢的影响

目的 观察慢性应激对阿尔茨海默病小鼠行为学和自由基的影响,并探讨逍遥散对此的干预作用.方法 将小鼠随机分为4组,即空白对照组、D-半乳糖组、复合模型组、逍遥散组.空白对照组给予0.9％生理盐水颈背部皮下注射,D-半乳糖组、复合模型组、逍遥散组均按120 mg/kg颈背部皮下注射D-半乳糖溶液,连续42 d.造模第29 d逍遥散组开始给予逍遥散13 g·kg-1·d-1灌胃,连续14 d;每日灌胃以后复合模型组和逍遥散组均给予慢性束缚应激刺激,每天6h.实验结束后对各组小鼠进行避暗试验和矿场试验,并于行为学检测后再检测自由基代谢指标超氧化物歧化酶（SOD）和丙二醛（MDA）的含量.结果 慢性束缚应激刺激导致阿尔茨海默病小鼠避暗试验中潜伏期缩短,错误次数增多;矿场试验中中央区域路程和总路程均减少,活跃度降低;自由基检测中SOD减少,MDA增多;逍遥散干预后小鼠行为学异常好转,SOD增多,MDA减少.结论 慢性束缚应激刺激能加重阿尔茨海默病;逍遥散能明显改善慢性束缚应激对阿尔茨海默病小鼠行为学的异常,并能减轻自由基损害.     

GSK3抑制剂氯化锂对脆性X综合征模型小鼠避暗行为的干预作用

目的探讨糖原合成酶激酶3(glycogen synthase kinase3,GSK3)抑制剂氯化锂对脆性X综合症小鼠模型的避暗行为的干预作用及机制。方法通过对30日龄脆性X综合症小鼠连续腹腔注射不同剂量氯化锂5 d,用药第4天和第5天进行避暗实验;同时用免疫印迹技术检测Fmr1 knockout(KO)及wild type(WT)小鼠的海马和皮层总GSK 3β和磷酸化GSK 3β(P-GSK3β)的变化。结果在避暗实验中,KO鼠与WT鼠,两者潜伏期及错误次数分别为(56±32)s,(83±24)s;(7±3)次,(3±2)次;免疫印迹实验结果:KO鼠皮层及海马P-GSK3β表达平均灰度值分别为69,63;WT鼠皮层和海马均为100。注射氯化锂后,KO鼠和WT鼠总GSK3β无明显改变,而KO鼠60 mg/kg,120 mg/kg,200 mg/kg组皮层P-GSK3β表达平均灰度值分别为:147,151,234;海马P-GSK3β分别为108,111,146,较空白组增多;P<0.05。WT鼠用氯化锂后,潜伏期和错误次数以及P-GSK3β表达变化无统计学意义。结论氯化锂能改善KO鼠的学习记忆能力,可能与氯化锂导致的P-GSK3β的表达增加有关,对脆性X综合征基因敲除小鼠有治疗作用。     

经颅电刺激运动诱发电位监测麻醉方法的建立

目的 对比相同麻醉深度下丙泊酚、七氟醚分别复合瑞芬太尼维持麻醉对经颅电刺激运动诱发电位(TES-MEPs)波幅和潜伏期的影响,以建立术中MEP监测时的适宜麻醉方法.方法 36例脊髓脊柱神经外科手术患者随机分为两组:丙泊酚麻醉组和七氟醚麻醉组,每组18例.对比麻醉诱导后30 min、60 min、90 min和120 min MEP的波幅和潜伏期.结果 两组病例麻醉期间血流动力学均维持稳定,组间脑电双频指数、呼气末二氧化碳分压和体温的差异均无统计学意义.丙泊酚组MEP所需阈刺激强度显著低于七氟醚组[(172±23)V:(217±42)V,P＜0.05].丙泊酚组各时间点上肢MEP波幅显著高于七氟醚组(P＜0.05),潜伏期显著短于七氟醚组(P＜0.05).两组内各时间点上肢MEP波幅和潜伏期差异无统计学意义.结论 在相同麻醉深度下,丙泊酚复合瑞芬太尼全凭静脉麻醉对MEP波幅的抑制作用显著优于七氟醚复合瑞芬太尼静吸复合麻醉方案,是用于脊髓脊柱手术TES-MEPs监测的适宜麻醉方法.     

长期口服氨氯地平对正常成年小鼠学习记忆的影响

目的探讨长期口服氨氯地平对正常成年小鼠学习记忆功能的影响,为氨氯地平的临床应用提供实验依据。方法选用成年小鼠,分为正常对照组及氨氯地平组,氨氯地平0.7～1.2mg/kg·d连续给药12w,通过Morris水迷宫、避暗、跳台行为学及脑胆碱系统测定,观察氨氯地平对正常成年小鼠学习记忆的影响。结果与正常对照组比,氨氯地平组小鼠1d至4d到达平台的潜伏期及游程、朝向角、游泳速度无明显改变,5d小鼠在2min内穿越平台的次数、在平台区的逗留时间、在平台区象限的逗留时间及平台区象限内的游程占总游程的百分比、平均速度、朝向角无明显变化(P>0.05);与正常对照组比,氨氯地平组小鼠2d避暗的错误次数及错误潜伏期也无明显变化(P>0.05);与正常对照组比,氨氯地平组小鼠1d、2d跳台的错误次数及2d的错误潜伏期没有明显改变(P>0.05);与正常对照组比较,氨氯地平组小鼠脑组织Ach含量明显增高(P<0.01)、AchE含量明显升高(P<0.05)。结论氨氯地平长期应用可能提高正常成年小鼠学习记忆功能。     

槲皮素对缺血缺氧损伤星形胶质细胞基因表达的影响

目的 应用高密度寡核苷酸(Oligo)基因芯片技术研究槲皮素对缺血缺氧损伤的星形胶质细胞基因表达的影响.方法 体外原代培养星形胶质细胞分为缺血缺氧组和缺血缺氧+槲皮素处理组.2组细胞厌氧培养4h后缺血缺氧+槲皮素处理组加入含50 μmol/L槲皮素的培养液,缺血缺氧组加入等量培养液,培养24 h后应用基因表达谱芯片筛选2组细胞表达差异的基因并用实时荧光定量PCR检测进行验证.结果 基因表达谱芯片分析显示缺血缺氧+槲皮素处理组与缺血缺氧组比较表达差异的基因共180个,其中上调基因49个,下调基因131个;实时荧光定量PCR结果 显示缺血缺氧+槲皮素处理组细胞与缺血缺氧组比较148个基因的表达发生变化,差异均有统计学意义(P＜0.05),其中上调基因34个,下调基因114个.实时荧光定量PCR与基因表达谱芯片结果 的符合率为82.2％(148/180).结论 基因表达谱芯片分析有助于从分子水平全面了解槲皮素对缺血缺氧的星形胶质细胞的作用机制,也为进一步研究槲皮素和星形胶质细胞在缺血缺氧脑损伤中的作用奠定了基础.     

咪达唑仑和丙泊酚诱导麻醉对继发性癫痫 术中皮层脑电图的影响

目的 探讨咪达唑仑与丙泊酚诱导麻醉对继发性癫痫术中皮层脑电图（ECoG）的影响。方法 2013年6月至2015年9月收治非功能区占位性病变继发性癫痫180例,根据诱导麻醉方法分为咪达唑仑组（60例;0.2 mg/kg）、丙泊酚组（60例;2 mg/kg）和咪达唑仑+丙泊酚组[60例;咪达唑仑（0.05 mg/kg）+丙泊酚（1 mg/kg）]。打开硬膜后行第一次ECoG监护,病灶切除后行第二次ECoG监护。结果 从麻醉诱导至第一次监护的时间为（110.3±15.9）min。第一次ECoG监护显示,咪达唑仑组暴发抑制（BS）波形发生率（68.3%,41/60）明显高于丙泊酚组（25.0%,15/60;PP<0.05）,后两组之间无统计学差异（>P>0.05）。麻醉诱导至第一次监护时间≤120 min时,咪达唑仑组BS发生率均明显高于丙泊酚组（P<0.05）和咪达唑仑+丙泊酚组（>P<0.05）;>120 min时,3组BS发生率无统计学差异（P>0.05）。第二次ECoG监护显示,咪达唑仑组、丙泊酚组和咪达唑仑+丙泊酚组BS发生率分别为5.0%（3/60）、3.3%（2/60）和3.3%（2/30）,3组之间无统计学差异（P>0.05）。结论 继发性癫痫患者术前镇静类诱导麻醉用药宜选用小剂量咪达唑仑联合丙泊酚。     

快速老化痴呆模型小鼠SAMP8学习记忆能力的增龄性变化

目的对快速老化痴呆模型小鼠SAMP8学习记忆能力的增龄性变化进行较系统的研究,为利用该模型进行其他研究提供实验依据。方法此实验选用1、4、8、12月龄的快速老化痴呆模型小鼠SAMP8,与同龄的正常老化小鼠SAMR1作对照,从老化度评分、避暗实验、Morris水迷宫实验和自主活动实验等方面观察了SAMP8小鼠学习记忆能力的增龄性变化。结果与对照组SAMR1相比,SAMP8小鼠随月龄增加老化度评分呈增高趋势,在8、12月龄的老化度评分值显著高于同龄对照组(P<0.05);避暗实验中,8、12月龄的SAMP8小鼠在电击24h后进入暗箱的潜伏期比同龄SAMR1小鼠显著缩短(P<0.05);Morris水迷宫实验中,1、4月龄SAMP8小鼠找到暗台的潜伏时间与同龄SAMR1小鼠相比差异无显著性,而8、12月龄SAMP8小鼠与同龄对照组相比,潜伏时间显著延长(P<0.05);从自主活动实验看,1、4、8月龄SAMP8小鼠单位时间内自主活动次数与同龄SAMR1小鼠相比无显著变化,而12月龄SAMP8小鼠与同龄对照组相比单位时间内自主活动次数显著减少(P<0.05)。结论SAMP8小鼠随月龄增长学习记忆能力逐渐减退;与同龄对照组相比,8、12月龄SAMP8小鼠出现明显衰老特征,表现出学习记忆能力明显低下,故可作为老化痴呆的动物模型用于痴呆有关研究。     

Free-operant avoidance of alcohol: An analogue study of alcohol aversion

An experimental analysis of the effects of peripheral electric shock on free-operant avoidance of alcohol was performed on five hospitalized alcoholics. A single-case design in which the aversive stimulus was systematically presented and withdrawn was used with each subject. There was little or no avoidance responding by any of the subjects in the initial baseline (no shock) condition. During conditioning, four of the subjects required relatively high shock intensities before making any avoidance responses. Removing and reinstating the shock contingency demonstrated that the shock exerted precise control over two subjects' avoidance behavior, some control over one subject and little or no control over the other two. After 100% avoidance was established, each subject continued to avoid alcohol even when the shock contingency was removed. Although there appeared to be some generalization of the avoidance to the subjects' natural environment, there are serious questions about the treatment potential of the procedure because of the high shock intensities that were necessary to condition the avoidance.     

Behavioral differences in the developing rat following postnatal anoxia or postnatally injected AF-64A, a cholinergic neurotoxin

Rat pups were submitted postnatally to one of two procedures: a 25-min exposure to 100% nitrogen or an i.c.v. bilateral injection of AF-64A, 2 nmol contained in 1-microliter saline. Throughout further development of either group, their performance in passive and active avoidance tests and in amphetamine-induced stereotype behavior was followed and compared. Both groups exhibited hyperactivity which persisted until 42 days of age in the anoxia group and beyond 120 days in the AF-64A group. Both groups were equally inferior to controls in the passive avoidance test, but only the anoxia group was inferior to controls in the active avoidance test. Amphetamine-induced stereotype behavior was much less pronounced in the anoxia group relative to AF-64A-treated rats or to controls. The results suggest that the lesion induced by the neurotoxin is more specific and less widespread than the one caused by anoxia.     

Avoidance learning in autoimmune mice

Previous studies have shown that autoimmune mice perform very poorly on active avoidance learning tasks. In the current studies, mice with lupus-like systemic autoimmunity were able to learn active, as well as passive, avoidance protocols with shock as reinforcement. Therefore, the behavioral deficits seen in active avoidance tasks are not a consequence of the use of electric shock. Rather, the current findings suggest that the inability of autoimmune mice to learn shock motivated responding is due to multiple performance factors, including shock level and properties of the testing apparatus.     

依托咪酯对无抽搐电休克治疗的影响

目的：探讨依托咪酯对丙泊酚诱导麻醉后无抽搐电休克（MECT）治疗运动发作时间不良的精神分裂症患者的疗效。方法：将67例行丙泊酚诱导麻醉下MECT治疗3次后，运动发作时间〈20s的精神分裂症患者随机分为两组，丙泊酚组33例，诱导麻醉仍给予丙泊酚，后续治疗电量较上次递增5％-100％；依托咪酯组34例，诱导麻醉换用依托咪酯，后续治疗电量较上次递增5％。两组均隔日治疗1次，共3次，治疗前及治疗2周，以阳性与阴性症状量表（PANSS）和韦氏记忆量表（WMS）评定其精神状况及记忆水平；比较两组患者运动发作时间、躁动谵妄发生率及疼痛发生率指标。结果：治疗后依托咪酯组运动发作时间、WMS评分、PANSS减分及躁动谵妄发生率均高于丙泊酚组，但疼痛发生率及电量指数显著低于丙泊酚组。结论：依托咪酯可提高丙泊酚诱导麻醉后MECT治疗运动发作时间不良患者的发作时间，减少注射疼痛，且较增加电量延长发作时间的方法有更少的认知损害，但须注意躁动谵妄问题。     

常压缺氧性脑损伤小鼠血清蛋白质表达谱的变化及意义

目的观察常压缺氧小鼠脑组织损伤之特点及其血清差异蛋白质表达,探寻常压缺氧性脑损伤的生物标志蛋白。方法60只雌性昆明小鼠,随机分为正常对照组和常压缺氧组,建立常压缺氧小鼠模型并按缺氧持续时间分为缺氧1周、2周和3周组,分别检测血清和脑组织中超氧化物歧化酶活性和丙二醛含量;应用弱阳离子交换芯片结合表面增强激光解析电离飞行时间质谱技术分析常压缺氧后不同时间血清蛋白质表达谱的变化。结果常压缺氧后小鼠血清超氧化物歧化酶活性逐渐降低,其中缺氧3周组与正常对照组之间差异有统计学意义（P〈0.01）;常压缺氧2周组小鼠血清丙二醛含量明显升高,与正常对照组和缺氧1周组比较差异有统计学意义（均P〈0.01）。常压缺氧后各亚组小鼠脑组织中超氧化物歧化酶活性与正常对照组比较差异无统计学意义（均P〉0.05）;常压缺氧后脑组织中丙二醛含量呈升高趋势,缺氧2周组和3周组与正常对照组比较差异有统计学意义（均P〈0.01）。弱阳离子交换芯片结合质谱分析共获得342个血清蛋白质峰,其中相对分子质量为3500、3578、3706、3516和4130等5个蛋白质峰的相对强度在缺氧性脑损伤后升高,与正常对照组相比差异有统计学意义（P〈0.05）;另有73个蛋白质峰分布于各缺氧组但在正常对照组中未检测到。这78个蛋白质峰中30个蛋白质峰于缺氧后≤2周表达,48个蛋白质峰于缺氧2周后表达。结论常压缺氧可引起小鼠脑组织损伤,使血清及脑组织中与氧自由基相关的酶活性发生改变,血清超氧化物歧化酶活性降低,血清及脑组织中丙二醛含量升高;并可改变血清蛋白质的表达谱。提示血清中出现的部分蛋白质可能为常压缺氧性脑损伤诱导的血细胞基因表达产物,在血清中所检测到的差异蛋白质以及缺氧后新出现的蛋白质可能是脑损伤的生物标志蛋白。     

Neuroprotection effects of retained acupuncture in neurotoxin-induced Parkinson's disease mice

The aim of this study was to investigate the role of retained acupuncture (RA) in neurotoxin-induced Parkinson’s disease (PD) mice. Male C57BL/6 mice were injected with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to induce the PD model. The mice were divided into four groups, namely, (1) normal; (2) MPTP + retained acupuncture (RA); (3) MPTP + electroacupuncture (EA); (4) MPTP + sham acupuncture (SA). After mice being manipulated with/without acupuncture at acupoints (Daling, PC 7), groups 2–4 were injected with MPTP (15 mg/kg/d). The mice were evaluated for behavioral changes, in terms of time of landing, after acupuncture treatment. The animals were sacrificed and their brains assayed for dopamine and its metabolites and tyrosine hydroxylase (TH) expression by using HPLC and immunohistochemistry/Western blotting, respectively. [¹²³I] IBZM-SPECT imaging between SA and RA groups were compared. The results showed that the time of landing of the three groups with treatment was significant longer than group 1 (normal) (4.33 ± 0.15 s). Nonetheless, group 2 (RA) (7.13 ± 0.20 s) had a shorter time of landing than group 4 (SA) (7.89 ± 0.46 s). The number of TH (+) neurons and the expression of TH proteins were significantly higher in the RA group than in the SA/EA groups. RA also increased the uptake of [¹²³I] IBZM into the triatum compared to the SA group. We conclude that RA possibly attenuates neuronal damage in MPTP-induced PD mice, which suggests RA may be useful as a complementary strategy when treating human PD.     

Facilitation of avoidance behaviour in mice chronically treated with heroin or methadone

Although the repercussion of chronic treatment with large amounts of opioids on cognitive performance is a matter of concern, the effects of opioid drugs on passive avoidance learning have been scarcely studied. Here, we analyzed the effects of prolonged administration of heroin and methadone, as well as the impact of suffering repeated episodes of withdrawal on fear-motivated learning using the passive avoidance test. Mice received chronic treatment (39 days) with methadone (10 mg/kg/24 h), associated or not with repeated withdrawal episodes, or with heroin (5 mg/kg/12 h). Our results show that, regardless of the type of treatment received, all mice displayed similar basal thermal nociceptive thresholds during 25 days of treatment. In the hot plate test, both methadone and heroin induced antinociception 30 min after drug administration. The analgesic effect was absent when measured 4 h after heroin and 12 h after methadone. Pain behavioural responses elicited by growing intensities of electric shock, applied on day 28th of treatment, were similar in all groups of mice. Our results indicate that chronic opioid treatment had promnesic effects on passive avoidance behaviour in mice, unrelated to changes in the nociceptive state.     

Effect of electrical convulsions on uridine labeling and activity pattern in nerve cells in mice

Male white mice were exposed to electroshock and then injected intravenously with 5-[3H]uridine immediately after the shock. After 5, 30, or 60 min or 6, 12, or 24 h, the mice were killed, microautoradiographs were prepared, and grains were counted in the cortex, hippocampus, and basal ganglia. The results of the grain counts were compared with grain counts in the cortex, hippocampus, and basal ganglia of mice exposed to anoxia for 25 s and then treated in the same manner as the first groups. After electroshock the grain count decreased to 25% of that in control animals in the hippocampus and to 50% in the cortex but was normal in the basal ganglia. The counts returned to normal values within 6 h in the hippocampus, and within 1 h in the cortex. After anoxia, the grain counts were normal in the cortex and hippocampus but increased in the basal ganglia. The distribution of cells with a high or low grain count in vertical and horizontal columns of the cortex in control and convulsion animals was analyzed. There were random variations from column to column in both control and convulsion animals. In some anatomic layers there were significantly different grain counts, indicating differences in functional activity.