TSPO激活剂YL-IPA08抗焦虑、抗抑郁作用及其机制研究

目的研究YL-IPA08的抗焦虑、抗抑郁作用,探讨其作用机制,并比较其与经典苯二氮类药物的作用特点与优势。方法采用放射配体竞争结合抑制实验检测受体亲和力;采用小鼠高架十字迷宫模型和大鼠Vogel饮水冲突模型抗焦虑抗焦虑行为效应,小鼠悬尾模型和大鼠强迫性游泳模型评价抗抑郁行为效应;并采用小鼠开场自发活动模型、戊巴比妥阈下剂量催眠模型、牵引模型、转杆模型、跳台模型,评价与CBR相关的潜在不良反应;采用ELISA法或放免法检测神经甾体水平;采用CCK8法检测细胞活性。结果 (1)受体结合实验证实:YL-IPA08与TSPO具有高亲和力(Ki=0.23 nmol·L-1),是对照化合物AC-5216(Ki=0.65 nmol·L-1)的2.8倍,且与CBR无结合。(2)药效学评价显示:YL-IPA08在小鼠悬尾模型(0.1～0.3 mg·kg-1,ig)和大鼠强迫性游泳模型(1～10 mg·kg-1,ig)上具有显著的抗抑郁作用,且可以被TSPO特异性拮抗剂PK11195(3 mg·kg-1,ip)所拮抗。YL-IPA08在小鼠高架十字迷宫模型(0.1～1 mg·kg-1,ig)和大鼠Vogel饮水冲突模型(3～6 mg·kg-1,ig)上具有抗显著的焦虑作用,而不能被CBR受体拮抗剂Flumazenil(10 mg·kg-1,ip)所拮抗。与苯二氮类抗焦虑剂地西泮(3 mg·kg-1,ip)不同,YL-IPA08不影响小鼠中枢兴奋性(0.3～30 mg·kg-1,ig),无镇静催眠效应(30～60 mg·kg-1,ig),对肌张力、运动平衡能力(0.3～100 mg·kg-1,ig)及记忆功能(0.3～30 mg·kg-1,ip)亦无明显影响。(3)机制研究发现:YL-IPA08(1～10 mg·kg-1,ig)可显著增加大鼠血清孕烯醇酮和孕酮的含量;YL-IPA08(0.1～2μmol·L-1)可显著促进体外培养星形胶质细胞合成分泌孕烯醇酮和孕酮的水平;YL-IPA08(0.03～3μmol·L-1)对皮质酮(200μmol·L-1)引起的星形胶质细胞损伤具有保护作用,该作用可被Trilostane所阻断。结论 YL-IPA08具有抗焦虑和抗抑郁作用,该作用可能与其高选择性激活TSPO,促进胶质细胞合成分泌神经甾体,保护神经细胞免受高浓度皮质酮损伤有关。YL-IPA08不产生明显的镇静、肌松、记忆障碍等中枢神经系统不良反应,较之地西泮具有明显的特色与优势。     

作用于外周苯二氮卓受体的新型抗焦虑剂

外周苯二氮卓受体位于线粒体上，调节胆固醇在线粒体膜上的转运速率，而胆固醇是线粒体合成类固醇的原料，合成的类固醇能调节GABAA受体的行为。外周苯二卓受体和适当的配体结合后，通过调节类固醇的合成间接地调节GABAA受体的行为，表现出明显的抗焦虑作用。此类化合物没有传统抗焦虑剂的肌松、镇静等副作用。本文介绍了外周苯二氮卓受体的生理生化机制及其在抗焦虑中的作用以及作用于外周苯二卓受体的几类新的抗焦虑化合物。     

黄芩苷对孤养小鼠的抗焦虑作用

目的研究黄芩苷(baicalin)对孤养小鼠的抗焦虑作用。方法制备孤养小鼠模型,应用小鼠高架十字迷路实验、小鼠明暗箱实验、小鼠孔板实验和隔离小鼠攻击实验考察黄芩苷对孤养小鼠的抗焦虑作用。结果黄芩苷可以对抗孤养小鼠在高架十字迷路实验、明暗箱实验和孔板实验中所表现出的焦虑样行为,在隔离小鼠攻击实验中表现抗攻击作用。结论黄芩苷对孤养小鼠有显著的抗焦虑作用。     

四氢孕酮和地西泮对暴露于架高十字迷宫小鼠的作用

目的比较孕酮的还原性代谢产物四氢孕酮和地西泮抗焦虑作用。方法C57小鼠腹腔注射四氢孕酮、地西泮或赋形剂 ,20min后观察在架高十字迷宫试验中的表现 ,测定其自发活动。结果腹腔注射四氢孕酮0.1mg·kg-1,明显缩短小鼠进入十字迷宫开臂的潜伏期[从(31.30±8.39)s减少到(8.80±6.00)s ,P<0.001] ,并显著增加进入十字迷宫开臂的次数(从1.20±0.42增到4.80±1.75,P<0.001)及在开臂的滞留时间占总时间的百分比 (从7.13 %增加到32.50 %,P<0.001)。而腹腔注射地西泮0.25mg·kg-1的抗焦虑作用弱于四氢孕酮。自发活动实验显示 ,0.5mg·kg-1地西泮明显减少小鼠自发活动 (P<0.01) ,而0.1mg·kg-1地西泮和0.25mg·kg-1 的四氢孕酮均不影响小鼠自发活动。结论试验结果提示神经甾体和地西泮对小鼠行为有不同的作用。四氢孕酮具选择性抗焦虑作用而不影响自发活动 ,可望成为地西泮抗焦虑的代用品     

谷维素和维生素B1增强地西泮的抗焦虑作用

目的研究地西泮、谷维素和维生素B1按1∶32∶8的比例合用后的抗焦虑作用,并同地西泮比较.方法采用大、小鼠高架十字谜宫模型和大鼠五甲烯四氮唑辨别效应对抗模型.结果在大、小鼠高架十字迷宫实验中,地西泮与谷维素和维生素B1合用后,大、小鼠在开放臂停留时间百分率明显增加,表现出明显的抗焦虑作用;而且其在开放臂停留时间百分率的增加,明显高于单独使用与其含量相等的地西泮的作用.在大鼠五甲烯四氮唑辨别效应对抗实验中,合用后对抗大鼠五甲烯四氮唑辨别效应的ED50值为23.4 mg·kg-1,其地西泮含量为0.6 mg·kg-1,远低于地西泮单方的ED50值3.5mg·kg-1.结论谷维素和维生素B1具有增强地西泮的抗焦虑作用.     

间氯苯哌嗪诱导的小鼠明暗箱焦虑模型 一种新的经济简便抗焦虑药筛选模型

目的　建立经济简便的抗焦虑药模型。方法　用间氯苯哌嗪 (mCPP)诱导产生焦虑 ,观察ddy小鼠和ICR小鼠在明暗箱的行为表现。结果　mCPP在sc 1～ 4mg·kg-1的剂量下即可显著降低ddy小鼠在明箱的活动次数 ,而对暗箱的活动次数影响不显著 ;mCPP在sc 2～ 10mg·kg-1的剂量下对ICR小鼠在明箱的的活动次数影响不明显 ;用地西泮对该模型进行验证 ,发现只用较小的样本量即可得出显著结果。结论　ddy小鼠可取代Wistar大鼠进行mCPP诱导焦虑的明暗箱模型 ,且简便易得 ,经济有效。     

组胺与扑尔敏对焦虑的影响

目的观察和分析组胺与扑尔敏对焦虑的影响。方法选用小鼠高架十字迷宫实验和小鼠明暗箱实验分别测定各组小鼠进入开放臂和封闭臂的总次数(OE+CE)、进入开放臂次数百分比(OE%)、开放臂停留时间比例(OT%)、探究次数(HT)和封闭臂后腿直立次数(RT)、明暗箱中的穿箱次数(TT)和在明箱停留时间的百分比(LT%)。结果剂量为4.44×l0-6g·kg-1的组胺(HA)能显著减少小鼠的OT%(P>0.05);4.00×l0-3g·kg-1的扑尔敏以及同剂量的扑尔敏加4.44×l0-6g·kg-1HA能显著增加小鼠的OE+CE、OE%、OT%、TT、LT%(P>0.01)。结论扑尔敏具有抗焦虑的作用,外周给予一定量的HA可以通过H1受体发挥促进焦虑的作用。     

作用于外周苯二氮受体的新型抗焦虑剂

外周苯二氮 受体位于线粒体上 ,调节胆固醇在线粒体膜上的转运速率 ,而胆固醇是线粒体合成类固醇的原料 ,合成的类固醇能调节GABAA 受体的行为。外周苯二氮 受体和适当的配体结合后 ,通过调节类固醇的合成间接地调节GABAA 受体的行为 ,表现出明显的抗焦虑作用。此类化合物没有传统抗焦虑剂的肌松、镇静等副作用。本文介绍了外周苯二氮  受体的生理生化机制及其在抗焦虑中的作用以及作用于外周苯二氮 受体的几类新的抗焦虑化合物     

2-芳基-3-吲哚取代乙酰胺类衍生物的合成及抗焦虑活性

目的　合成2-芳基-3-吲哚取代乙酰胺类化合物,从中筛选有抗焦虑作用而无镇静、肌松等副作用的活性化合物。方法　由取代苯和琥珀酸酐经傅-克反应得到取代苯甲酰基丙酸,取代苯甲酰基丙酸和氯甲酸乙脂生成混合酸酐,再和相应的胺反应得到取代酰胺,取代酰胺和取代苯肼经费歇尔反应得到目标化合物。结果　得到新化合物20个。结论　初步受体结果表明,多数化合物均与外周苯二氮受体有较强结合,在小鼠高架十字迷宫试验中发现一些化合物有明显的抗焦虑作用,且不能拮抗印防己毒素(PTX)诱发的惊厥作用,显示无镇静作用     

核黄素四丁酸酯抗焦虑作用的药效学研究

目的考察核黄素四丁酸酯(RTB)的抗焦虑作用。方法以地西泮和丁螺环酮为阳性对照药,采用小鼠高架十字迷宫和新型食物消耗实验,评价RTB抗焦虑作用;采用转棒实验测定RTB对小鼠肌肉协调运动能力的影响。结果与空白对照组比较,RTB不影响小鼠在高架十字迷宫中的入臂总次数,但在20和40 mg.kg-1的剂量下明显增加开臂进入次数百分比和在开臂中滞留时间百分比;在新型食物消耗实验中,RTB在剂量为40和80 mg.kg-1时,明显增加小鼠单位体重的食物消耗量;转棒实验中,与阳性药物比较,RTB组明显降低了小鼠的跌落率。结论 RTB具有抗焦虑的作用,且不影响小鼠的活动能力和肌肉协调性。     

Amphetamine-induced anxiety-related behavior in animal models

The purpose of this study was to examine the anxiety-related effects of acute and repeated amphetamine administration using the elevated plus maze (EPM) and light/dark box tests in mice. D-amphetamine (2 mg/kg ip, 30 min after injection) had a significant anxiogenic effect only in the EPM test, as shown by specific decreases in the percentage of time spent in the open arms as well as in the percentage of open arm entries. Tolerance to this anxiogenic action developed after 8 days of daily d-amphetamine administration (2 mg/kg, ip). An anxiolytic effect was observed after the ninth injection, i.e. there were specific increases in the percentage of time spent in the open arms and in the percentage of open arm entries. L-type voltage-dependent calcium channel antagonists: nimodipine (5, 10 and 20 mg/kg, ip), flunarizine (5, 10 and 20 mg/kg, ip), verapamil (5, 10 and 20 mg/kg, ip), and diltiazem (5, 10 and 20 mg/kg, ip) were also injected prior to an acute low dose of d-amphetamine or to each injection of subchronic d-amphetamine. Our results revealed that calcium channel blockers dose-dependently attenuated both an anxiogenic effect of d-amphetamine and the development of tolerance to this effect. Our results suggest that neural calcium-dependent mechanisms are involved in the anxiety-related responses to acute and subchronic amphetamine injection that may lead to addiction relapse in human users.     

Prophylactic and therapeutic effects of acute systemic injections of EMD 281014, a selective serotonin 2A receptor antagonist on anxiety induced by predator stress in rats

We examined the effect of the selective serotonin 2A (5-HT(2A)) receptor antagonist 7-[4-[2-(4-fluoro-phenyl)-ethyl]-piperazine-1-carbonyl]-1H-indole-3-carbon itrile HCl (EMD 281014) [Bartoszyk, G.D., van Amsterdam, C., Bottcher, H., Seyfried, C.A., 2003. EMD 281014, a new selective serotonin 5-HT2A receptor antagonist. Eur. J. Pharmacol. 473, 229-230.] on change in affect following predator stress. Predator stress involved a 5 min unprotected exposure of rats to a domestic cat. Behavioral effects of stress were evaluated with hole board, plus maze, light/dark box and acoustic startle tests 1 week after stress. Predator stress increased anxiety-like behavior in the plus maze, light/dark box, and elevated response to acoustic startle. EMD 281014 (0.001, 0.01, 0.1, 1 or 10 mg/kg) and vehicle injection (ip) occurred either 10 min after predator stress (prophylactic testing), or 90 min prior to behavioral testing for the effects of predator stress (therapeutic testing 1 week after predator stress). In prophylactic testing, EMD 281014 prevented stress potentiation of startle in a dose dependent manner, though the most effective doses were midrange (0.01 and 0.1 mg/kg). Prophylactic administration of EMD 281014 also prevented stress-induced increase of open arm avoidance in the plus maze in a clear dose dependent manner (from 0.01 mg/kg onward). In therapeutic testing, EMD 281014 had no clear drug dependent effects on stress elevation of startle or on behavior of stressed rats in the elevated plus maze. Finally, EMD 281014 did not block the effects of stress on behavior in the light/dark box when given prophylactically or therapeutically. Findings implicate 5-HT(2A) receptors in initiation of some but not all lasting changes in anxiety-like behavior following predator stress. Potential clinical significance of findings are discussed.     

缬草素抗焦虑作用及其机制的初探

目的研究缬草素的抗焦虑作用,并分析其可能的作用机制。方法以大鼠高架十字迷宫实验和开场实验为动物模型,考察缬草素的抗焦虑活性;用酶联免疫吸附法(ELISA)测定大鼠血清皮质酮的水平。结果缬草素(10mg.kg-1)能增加大鼠在开臂内运动时间和次数百分率,并且提高大鼠在开场中央区的次数,在大鼠高架十字迷宫与开场模型上显示出抗焦虑作用;酶联免疫吸附法表明:缬草素能降低大鼠血清皮质酮水平。结论缬草素具有明显的抗焦虑作用,机制可能与调节下丘脑-垂体-肾上腺轴功能有关。     

Anxiolytic activity of pyridoindole derivatives SMe1EC2 and SMe1M2: behavioral analysis using rat model

Anxiety and mood disorders have become very significant affections in the last decades. According to WHO at least one mental disease occurred per year in 27% of EU inhabitants (more than 82 mil. people). It is estimated that by 2020, depression will be the main cause of morbidity in the developed countries. These circumstances call for research for new prospective drugs with anxiolytic and antidepressive properties exhibiting no toxicity and withdrawal effect and possessing beneficial properties, like antioxidant and/or neuroprotective effects. The aim of this study was to obtain information about psychopharmacological properties of pyridoindole derivatives SMe1EC2 and SMe1M2, using non-invasive behavioral methods in rats.The battery of ethological tests (open field, elevated plus-maze, light/dark box exploration, forced swim test) was used to obtain information about anxiolytic and antidepressant activity of the pyridoindole derivatives. The substances were administered intraperitoneally 30 minutes before the tests at doses of 1, 10 and 25 mg/kg.In the behavioral tests, SMe1EC2 was found to exert anxiolytic activity in elevated plus maze with no affection of locomotor activity. The highest dose of SMe1M2 increased the time spent in the lit part of the Light/Dark box, however this result was influenced by inhibition of motor activity of the rats. Similar findings were observed also in elevated plus-maze, although these results were not statistically significant.In conclusion, from the results of our study it is evident that both pyridoindoles acted on the CNS. In the highest dose, SMe1M2 was found to possess rather sedative than anxiolytic or antidepressant activity.     

Reduction in the anxiolytic effects of ethanol by centrally formed acetaldehyde: the role of catalase inhibitors and acetaldehyde-sequestering agents

Rationale  Considerable evidence indicates that brain ethanol metabolism mediated by catalase is involved in modulating some of the behavioral and physiological effects of this drug, which suggests that the first metabolite of ethanol, acetaldehyde, may have central actions. Previous results have shown that acetaldehyde administered into the lateral ventricles produced anxiolysis in a novel open arena in rats. Objectives  The present studies investigate the effects of centrally formed acetaldehyde on ethanol-induced anxiolysis. Materials and methods  The effects of the catalase inhibitor sodium azide (SA; 0 or 10 mg/kg, IP) on ethanol-induced anxiolysis (0.0, 0.5, or 1.0 g/kg, IP) were evaluated in CD1 mice in two anxiety paradigms, the elevated plus maze and the dark/light box. Additional studies assessed the effect of the noncompetitive catalase inhibitor 3-amino-1,2,4-triazole (AT; 0.5 g/kg, IP) and the acetaldehyde inactivation agent d-penicillamine (50 mg/kg, IP) on the plus maze. Results  SA reduced the anxiolytic effects of ethanol on several parameters evaluated in the elevated plus maze and in the dark/light box. In the plus maze, AT completely blocked and d-penicillamine significantly reduced the anxiolytic properties of ethanol. Conclusions  Thus, when cerebral metabolism of ethanol into acetaldehyde is blocked by catalase inhibitors, or acetaldehyde is inactivated, there is a suppressive effect on the anxiolytic actions of ethanol. These data provide further support for the idea that centrally formed or administered acetaldehyde can contribute to some of the psychopharmacological actions of ethanol, including its anxiolytic properties.     

Protective effect of Withania somnifera dunal root extract against protracted social isolation induced behavior in rats

This study investigated the effect of Withania somnifera Dunal (WS) root extract and diazepam in social isolation induced behavior such as anxiety and depression in rats. Rats were isolated for 6 weeks and the assessment of changed behavior were done on elevated plus maze (EPM) and forced swim test (FST). Isolation reared rats spent less time into the open arms on EPM and significantly increased immobility time in FST compared to group housed rats. WS (100, 200 or 500 mg/kg, oral) and diazepam (1 or 2 mg/kg, ip) dose dependently increased the time spent and entries into the open arms on EPM test and showed the anxiolytic activity. Subeffective dose of WS (50 mg/kg, oral) potentiated the anxiolytic action of diazepam (0.5, 1 or 2 mg/kg, ip). WS (100, 200 or 500 mg/kg, oral) also reduced the immobility time in FST, thus showed antidepressant effect in both group housed and social isolates. The investigations support the use of WS as a mood stabilizer in socially isolation behavior in Ayurveda.     

孕酮对大鼠吗啡位置偏爱效应及中枢单胺递质水平的影响

目的观察孕酮对于吗啡所致奖赏效应及脑内单胺类神经递质水平的影响。方法采用大鼠条件性位置偏爱(CPP)模型,高效液相色谱-电化学法测定大鼠伏隔核及腹侧被盖区内去甲肾上腺素(NE)、多巴胺(DA)和5羟色胺(5HT)的含量。结果吗啡(5mg·kg-1)可诱导大鼠产生稳定的CPP效应;孕酮(5、20mg·kg-1)本身不产生CPP效应,但能抑制吗啡的CPP效应。与对照组比较,吗啡CPP形成时,伏隔核内NE和DA的水平明显升高(P<0.01)。与吗啡组比较,合用5mg·kg-1或20mg·kg-1孕酮均可使伏隔核内DA水平下降(P<0.01,P<0.05);合用20mg·kg-1孕酮还可使伏隔核内的NE水平下降(P<0.01)。结论孕酮可有效抑制吗啡的CPP效应,其机制可能与降低伏隔核内DA及NE的水平有关。     

褪黑素镇痛的相关机制

目的　探讨褪黑素 (MT)镇痛作用与内源性阿片肽、去甲肾上腺素能神经系统及钙通道的关系。方法　大鼠和小鼠痛阈的测定采用热板法 ;β 内啡肽 (β EP)的测定采用放免法 (RIA)。结果　松果腺切除后d 8大鼠痛阈的昼夜节律性消失 ,icvMT 0 2 5mg·kg-1可出现明显的镇痛作用 ;ipMT(10 0mg·kg-1) 1h后下丘脑、垂体 β EP含量均明显降低 ;ip利血平 (3mg·kg-1)可使MT镇痛作用消失 ,而sc酚妥拉明 (10mg·kg-1)可减弱MT的镇痛作用 ;CaCl2 (2 30mg·kg-1)与MT(40mg·kg-1)合用时 ,可使MT的镇痛作用减弱 ,EGTA (180mg·kg-1)及维拉帕米 (15mg·kg-1)则可使之加强。结论　MT的镇痛作用可能与内源性阿片肽、去甲肾上腺素能神经系统及Ca2 + 通道等有关     

川芎嗪对吗啡依赖大鼠戒断综合征的抑制作用

目的··:观察川芎嗪对吗啡依赖大鼠戒断综合征的影响。方法··:采用剂量递增方式皮下注射吗啡5d的大鼠,经过川芎嗪处理30min后,用纳洛酮催促,在停用吗啡后不同时间里观察评定戒断症状分值。结果··:川芎嗪明显减轻戒断症状和体重下降。结论··:川芎嗪能明显抑制吗啡依赖大鼠的戒断反应,其作用机制可能与阻断细胞钙内流有关。     

Impaired action of anxiolytic drugs in mice deficient in cannabinoid CB1 receptors

The role of cannabinoid CB(1) receptors in the action of anxiolytics was examined. Deletion of CB(1) receptors resulted in increased anxiety-like behaviours in light/dark box, elevated plus maze and social interaction tests. Mutant mice presented basal low corticosterone concentrations and low proopiomelanocortin gene expression in the anterior lobe of the pituitary gland compared to wild-type mice. Ten minutes of restraint stress resulted in a twofold increase in corticosterone concentrations in the plasma of mutant mice, compared to wild-type mice. Bromazepam (50 or 100 microg/kg) markedly increased the time spent in light area in wild-type animals, though both doses were without effect in mutant mice. Administration of buspirone (1 or 2 mg/kg) produced anxiolytic effects in wild-type mice. In contrast, only the highest dose of buspirone had anxiolytic results in mutant mice. Our findings reveal that CB(1) receptors are involved in the regulation of emotional responses, and play a pivotal role in the action mechanism of anxiolytics. They suggest that alterations in the functional activity of the CB(1) receptor may be related to the emergence of anxiety disorders, and may affect treatment with anxiolytics.