安神方抗焦虑作用的疗效观察及相关机制研究
|
摘要
进入21世纪以来,随着社会发展进程的加快竞争日趋激烈,生活、学习、工作等压力也随之不断加大,人们在不同程度上普遍感受到了精神压力,联合国世界卫生组织预测指出,精神病学以及心理卫生将越来越受到人们的重视。据统计在10利,造成社会最沉重负担的疾病中,精神疾病占了4种,精神疾病的总负担占全部疾病负担的1/4,精神医学正在逐步受到医学同行及社会的关注并被赋予新的认识。现在人们对精神健康的关注也从以往的重型精神障碍逐渐向轻型转变。
焦虑是精神障碍五大症状群中常见的一种,焦虑障碍作为神经症的一个独立的亚型,其发病率正呈现逐年上升趋势。焦虑障碍又称焦虑症,其主要有两种临床形式--广泛性焦虑障碍和惊恐障碍。广泛性焦虑障碍(generalized anxiety disorder, GAD)又称慢性焦虑症,其主要表现为对实际并不存在的威胁或危险以及身体健康过分的担心、紧张和害怕,并伴有口干、胃肠不适、恶心、腹泻、呼吸困难、心悸、尿频尿急、男性阳痿、女性月经不调、出汗、面色潮红及肌肉紧张、手颤、头痛、坐立不安等症状,另外还伴有注意力不集中、记忆力减退、睡眠障碍等症状。
随着我国经济发展和社会文明的进步,人们早已消除了对精神疾病的狭隘误解,正确的认识使得人们对自我心理健康的关注程度有了较大提高,因觉察到自身焦虑症状而主动就医的人数越来越多,焦虑障碍已经成为综合医院门诊、心理咨询和治疗机构中常见的一类疾病。越来越多患有焦虑症状的人群希望能够得到及时、有效的诊断和治疗,为此,全球各国医务工作者正在积极开展对焦虑障碍发病机制以及预防治疗方案等方面的研究。
目前,现代医学在对焦虑障碍的发病机制研究方面已经取得了很大进展,药物治疗方面也已具备了较为完善的理论。临床治疗多采用以地西泮为代表的苯二氮卓类药物和丁螺环酮等非苯二氮卓类药物,前者在使用初期起效很快但常伴有嗜睡、便秘、肌松、记忆力减弱等副作用,后者虽副作用较少,但起效通常较慢,另外该类药物由于容易形成依赖和耐受以及戒断反应等因素,故也在一定程度上限制了应用。因此,寻找疗效确切、副作用小的抗焦虑药物就成为了目前抗焦虑研究领域的重要课题。
我们的课题研究首先通过收集查阅大量相关文献,在学习借鉴现代医学对焦虑障碍的研究现状之后,找到了焦虑障碍与中医学情志病之间的相关性和共同点,并在总结了中医学历代、各学派学者对情志病的调治方法的基础之上,在博士生导师、知名中医陈宝田教授指导下,确立了清热除焦、调和营卫、安神定志的治疗原则,制定了安神方(ASF)的组方用药。教授结合个人近50年临床经验,在安神方中配伍运用柴胡、酸枣仁、桂枝、龙骨、牡蛎等药物,用于治疗焦虑症见焦躁易怒、胸胁胀闷、惊悸不安、失眠健忘、肢体困乏、口苦咽干、舌红脉弦数的患者。此次研究通过观察安神方治疗广泛性焦虑障碍(GAD)的临床疗效、并进一步通过动物试验探讨安神方治疗焦虑障碍的药效学及相关作用机制。旨在充分发挥祖国医学整体观念、辨证论治这些优势特点的同时,结合现代医学的实验方法,更好的解释其作用效果及机理,为更好的治疗焦虑障碍探索铺就出一条有效之路。
一、安神方治疗广泛性焦虑障碍(GAD)的疗效观察
1.目的：通过临床观察探讨安神方治疗广泛性焦虑障碍(GAD)的临床效果,观察症侯及临床整体疗效以及收集整理相关检查所得的结果,客观的评价安神方治疗GAD的临床疗效。
2.方法：
2.1病例与分组：将符合纳入标准的120例GAD入组患者随机分为治疗组和对照组,每组60例；
2.2用药：治疗组口服“安神方”(由南方医院中药房提供,方剂组成为：柴胡25g,桂枝10g,黄芩10g,党参15g,白芍15g,生龙骨30g,生牡蛎30g,炒枣仁15g,远志10g,夜交藤30g,甘草10g等),每日1剂,分2次,晨起空腹及晚睡前30min各服一次；对照组口服地西泮(由天津利生制药股份有限公司提供,2.5g/片,100片/瓶)口服,1片/次,晨起空腹及晚睡前30min各服一次。两组均以连续服药4周为1个疗程。治疗期间不同用其他抗焦虑及镇静安眠药。两组均于诊疗中同时注意对患者进行必要的心理疏导,避免情绪波动。
2.3改变指标：分别于治疗前及治疗后1、2、4周末各进行一次汉密尔顿焦虑量表(HAMA)评定。以HAMA减分率(T)作为疗效评定指标, T=(治疗前总分-治疗后总分)/治疗前总分×100%。T小于25%为无效,25%～49%为好转,50%～75%为显著好转,大于75%为临床痊愈；总有效率=(痊愈+显著+好转)/总例数×100%。
2.4临床整体疗效、不良反应及安全性判定：以汉密尔顿焦虑量表(HAMA)和副反应量表(TESS)评价疗效和不良反应及其程度。并于治疗前后检查：体温,心率,血压,呼吸；血、尿、便常规；肝、肾功能；心电图等项目,以此对两组服药后的不良反应以及药物的安全性进行全面的评价。
2.5统计学方法：采用统计软件SPSS13.0进行统计学处理和分析。试验数据计量资料采用均数加减标准差(X±S)表示。治疗前后剂量资料比较采用配对样本t检验；两组间计量资料比较采用独立样本t检验；治疗后组间症候疗效比较采用协方差分析；等级资料分布情况采用秩和检验及非参数检验。计数资料以百分率表示,采用卡方检验。所有统计数据均以P<0.05作为有显著性差异的界限。
3.结果：治疗4周后,两组HAMA分值均较治疗前有显著差异(P均<0.01),两组间比较差异无统计学意义(P>0.05)。治疗组与对照组TESS量表评分比较差异有统计学意义(P<0.05)。
4.结论：初步确认了安神方治疗GAD安全、有效且不良反应较小,值得进一步研究推广。
二、安神方抗焦虑作用的药效学研究
1.目的：选用大鼠高架十字迷宫试验模型(EPM),观察安神方的抗焦虑作用。
2.方法：将60只健康雄性Wistar大鼠随机分为6组(10只/组)：正常对照组、模型组、地西泮组(DZP)和安神方(ASF)高、中、低剂量组。采用慢性情绪应激的方法建立焦虑大鼠模型,同时给予药物干预,以国际公认的高架十字迷宫试验对其进行药效行为学评价。正常对照组不接受任何处理,自由饮水和摄食。其余各组定时喂水训练7天,之后开始应激试验,在上述两个时间段内给予不确定空瓶刺激,维持1天1次或2次,持续2周。在不确定空瓶刺激期间,地西泮组给予1mg/kg灌胃,模型组每天给予等容积生理盐水,ASF高、中、低剂量组每天剂量分别为：40g/kg、20g/kg、10g/kg。于第21天,ASF组末次给药2h、模型组及DZP组末次给药0.5h后,进行高架十字迷宫行为学测定,大鼠均提前2h进入测试实验室适应环境。
3.仪器：大鼠高架十字迷宫(EPM)采用国际通用的方法制作。
4.统计学方法：试验数据采用SPSS13.0统计软件进行统计分析,组间比较采用单因素方差分析,多重比较采用SNK法。试验数据均以均数加减标准差(x±s)表示,P<0.05为有统计学意义。
5.结果：与正常对照组比较,模型组大鼠进入开臂的时间百分率(OT%)次数百分率(OE%)均明显减少；DZP组与ASF高、中、低剂量组大鼠进入开臂的时间百分率(OT%)、次数百分率(OE%)均较模型组显著上升,其中以ASF中剂量组上升最明显。
6.结论：行为学试验结果显示：DZP组、ASF高、中、低剂量组同模型组比较, EPM中大鼠OE%和OT%值均升高(P<0.05),尤其是ASF中剂量组升高最明显(P<0.01),其作用效果与DZP相当,这进一步证实了ASF对大鼠焦虑状态的干预作用,而且以中剂量给药干预效果最好。
三、安神方抗焦虑作用的形态学及相关机制研究
1.目的：观察安神方(ASF)的抗焦虑作用,并初步研究其机制。
2.方法：①.将所取雄性Wistar大鼠随机平均分为6组：正常对照组、模型组、DZP组和ASF高、中、低剂量组；
②.采用慢性情绪应激的方法建立焦虑大鼠模型,同时给予药物干预,以国际公认的高架十字迷宫模型(EPM)试验对其行为学进行评价；
③.行为学观察后,将正常对照组、模型组、DZP组及ASF高、中、低剂量组大鼠立即麻醉处死、断头,于冰台上剥离大鼠的海马组织。
④.在光学显微镜下采用HE染色法观察大鼠脑组织内神经元形态结构的改变和胶质细胞的增生情况；用尼氏染色法观察神经元受损和尼氏体的丢失情况。
⑤.用免疫组化法观察ASF对焦虑大鼠脑组织内氨基酸类神经递质--Y-氨基丁酸(GABA)和谷氨酸(Glu)含量的影响；
⑥.用Western bolt法观察ASF对焦虑大鼠脑组织内氨基酸类神经递质受体-GABAA 受体(GABAARα1)和Glu受体(N-甲基-D-天冬氨酸受体1即NMDAR1)表达的影响；
⑦.用荧光定量PCR法观察ASF对焦虑大鼠脑组织内氨基酸类神经递质受体-GABAA受体(GABAARα1)和Glu受体(N-甲基-D-天冬氨酸受体1即NMDAR1)基因表达的影响。
3.仪器：MilliQS纯水／超纯水系统、6219型PH计、BCD-196DT冰箱、CR703恒温振荡器、SFG-02B电热恒温干燥箱、TS-1脱色摇床、Eoipse Ti-s倒置显微镜、MI-22FOEGC微波炉、RF2000荧光检测器、PHS-2C精密酸度计、Avanti30冷冻离心机、BA110S电子天平、TiThermocyclerRea-timepCR仪、CA-92-2垂直层流洁净工作台、DYCP水平电泳槽、DYY6B型稳压稳流电泳仪、sigma3K30超速冷冻离心机、MX3005P荧光定量PCR仪及分析软件MXPro4.01
4.试剂：PBS缓冲液(ph7.2—7.4)、高效切片石蜡、中性甲醛、无水酒精、二甲苯、浓盐酸、苏木精、伊红染、中性树脂、50ml离心管、普通玻片、盖玻片、玻片盒、玻片架、谷氨酸(Glu)购自美国SIGMA,公司批号分别为43907190；Y-氨基丁酸(GABA)购自美国SIGMA公司,公司批号116K5005；乙腈购自美国TEDIA公司；GABAARal兔IgG多克隆抗体购自美国santa curz公司；N-甲基-D-天冬氨酸受体KNMDAR1)兔IgG多克隆抗体购自美国santa curz公司;2mlPCR荧光管购自Axgen公司；Nalater:AMBION公司；2000bpDNALadder:广州瑞真生物科技有限公司；焦碳酸二乙醋(DEPC) treated water:大连TaKaRa宝生物公司；Trizol:Invitrogen公司、RNA反转录试剂盒：Takara公司、SyBrI荧光定量试剂盒购自Gene copoeia公司、琼脂糖购自Sigma公司,其余试剂均为进口及国产分装试剂。
5.统计学方法：试验数据采用SPSS13.0统计软件进行统计分析,组间比较采用单因素方差分析,多重比较采用SNK法。试验数据均以均数加减标准差(x±s)表示,以P<0.05为有统计学意义。
6.结果：
①.正常对照组：海马CA3区细胞排列整齐,细胞核圆而大,核仁清晰可见,锥体细胞排列整齐,层次紧密,尼氏体深染；模型组：海马CA3区锥体细胞排列紊乱,神经细胞出现明显的病理变化,染色质溶解,部分细胞脱失,胞体固缩呈多角形或不规则形,胞质浓缩核膜界限不清,锥体细胞排列松散,细胞缺失,尼氏体浅染甚至溶解；DZP组：海马CA3区锥体细胞排列比较整齐,少部分神经细胞出现病理变化,少部分细胞染色质溶解,少部分胞体固缩呈不规则形,锥体细胞排列部分松散,锥体细胞与中剂量组相当,细胞部分缺失；ASF低、中、高剂量组：海马CA3区锥体细胞排列比较整齐,神经细胞部分出现病理变化,部分细胞染色质溶解,部分细胞脱失,胞体固缩呈不规则形。其中中剂量组细胞形态修复最好,ASF低剂量组锥体细胞排列比较整齐,比模型组锥体细胞明显增多,细胞部分缺失,尼氏体浅染；ASF中剂量组锥体细胞排列比较整齐,锥体细胞与高剂量组相当,细胞部分缺失,尼氏体染色较高剂量组深染,与DZP剂量组相当；ASF高剂量组锥体细胞排列比较整齐,比低剂量锥体细胞有所增多,细胞部分缺失,尼氏体浅染。
②.模型组大鼠海马组织中Glu的表达量较正常对照组显著上升,GABA的表达量明显下降;地西泮组和安神方中剂量组同模型组比较,大鼠海马组织中Glu的表达量显著降低和GABA的表达量明显提高,安神方中剂量组与地西泮组比较差异不显著；
③.模型组与正常对照组比较,GABAA受体表达减少,Glu受体表达增多；地西泮组和安神方高、中、低剂量组分别同模型组进行比较,Glu受体的表达水平均降低,GABAA受体的表达水平均提高；安神方中剂量组与地西泮组比较无明显差异。
④.模型组与正常对照组比较,GABAA受体基因减少,Glu受体基因增多；地西泮组和安神方高、中、低剂量组分别同模型组进行比较,Glu受体基因水平均降低,GABAA受体受体基因水平均提高；安神方中剂量组与地西泮组比较无明显差异。
7.结论：安神方具有抗焦虑的作用,具体表现在：
①可以修复焦虑模型大鼠中枢神经系统中海马区出现的病理性变化；
②可以提高大鼠中枢神经系统中GABA的含量,降低Glu的含量；
③可以增加大鼠中枢神经系统中GABA受体的表达,减少Glu受体的表达；
④可以增加大鼠中枢神经系统中GABA受体基因的表达,减少Glu受体基因的表达。
以上研究结果进一步证实了安神方对大鼠焦虑状态的干预作用,而且以中剂量给药干预效果最好。
While advances in science and technology have brought huge amounts of benefits to modern societies, they have also caused numerous side effects that increasingly threaten people's heath. The modern world is fast-paced and filled with fierce competition at a global level, and as a result, millions of people are under more pressure than ever before, and most would have to face deteriorated working conditions such as longer working hours, the requirement of higher level of concentration, insufficient physical exercises, etc.. World Health Organization (WHO) reported that nowadays people pay more and more attention to mental health and psychological welbeing [REF]. It also pointed out that among the10diseases that are most costly to taxpayers,4are classified as mental illness. In addition, the treatment of mental diseases costs a quarter of the total budget on health. These facts have shown that mental health has brought an increasing amount of attention, not only from medical practitioners, but also governments and their citizens.
Anxiety disorder is one of the biggest type of mental illness and it shows an increasing incidence in recent years. Anxiety disorder covers several different forms of abnormal and pathological fear and anxiety. It is divided into Generalized Anxiety Disorder (GAD) and Panic Disorder (PD), where GAD is a common chronic disorder characterized by long-lasting anxiety that is not focused on any one object or situation. Those suffering from GAD are frequently accompanied by physiological symptoms such as dry mouth, upset stomach, nausea,diarrhea, dyspnea, palpitations, frequent urination, impotentia,menstrual disorder, sweating, flushed face,①muscle tension and spasms.headache, short attention span, hypomnesis and sleep disorder. In China, along with the economic development in recent years, people have realized the importance of mental health. As people become more active in seeking medical advice on symptoms of anxiety disorder, hospitals and specialized clinics have admitted an increasing number of patients with such disease, which has driven a surge in research in the physiology of anxiety disorder and its treatment.
Currently, modern medicine not only has made great progress in the pathogenesis of anxiety disorders, but also has a sound theoretical in drug treatment. There are two classes of drugs being mostly used in clinical treatment, benzodiazepine drugs such as Diazepam and non-benzodiazepine drugs such as Buspirone. The former ones effect quickly in the beginning, but often accompanied by lethargy, constipation, muscle relaxants, memory weakened, and other side effects. The latter ones, with fewer side effects, effect slower. Moreover due to the easy formation of dependence and tolerance, and withdrawal reactions, to some extent limits the application of this classes of drugs. Therefore to find out the drugs with efficacy and fewer side effects, has become an important topic of the current anti-anxiety research areas.
After accessing large amounts of literatures and leaning from anxiety disorders researches of modern medicine, our subject of study finds out correlations between anxiety disorders and these disorders of Chinese medicine. Under the guidance of the doctoral mentor, the renowned Chinese Medicine Professor Chen Baotian, on the basis of summarizing,we establish three treatment principles, and set up AnShenfang combined with almost50years of clinical experiences of professor Chen, we use chaihu,guizhi,longgu,muli and others medicines in AnShenfang for the treatment of anxiety. This study observes the clinical efficacy of AnShenfang in treatment of Generalized Anxiety Disorder (GAD), and further with animal experiments, investigates the pharmacodynamics, morphology and the mechanism of actions of AnShenfang. Aimed to give full play to the concepts of Chinese Medicine and the advantages of treatment based on syndrome differentiation,combined with modern medical experiments, our study is designed to better explained the role of effects and mechanisms of AnShenfang, in order to explore and pave out an effective way for better treatment of anxiety disorders.
chapter1. Observation of AnShenFang in Treating Generalized Anxiety Disorder
l.Objective:To investigate the efficacy of AnShenFang(ASF) for patients with general anxiety disorder (GAD). To objectively estimate the therapical effect of "AnShenfang" in treating GAD through the clinical overall curative effect as well as the finding.
2.Methods:Clinical curative effect observation:120cases according with the diagnostic standard were equally divided into (ASF) treatment group and western medicine treatment(DZP) control group at random.
During treatment:The patients were entered a4-week randomized contralled triad.60patients of control group were given DZP and other60of treatment group were given ASF.other sedative and anxiolytic medicines should be abstinent.Mental nursing is necessary besides the therapy of during treatment
The HAMA were measured before and after3,6,12months of the treatment. The results were compared with ASF treatment group and western medicine treatment control group.
Assess the benefits and adverse reaction of ASF and DZP in the treatment of GAD, according to the level of Hamilton Anxiety Scale (HAMA) and Treatment Emergent Symptom Scale (TESS)Before and after treatment, temperature, heart rate, blood pressure, renal and hepatic funtion, plasmal electrolyte and electrocardiogram were assessed and compared.
3. Results:After4weeks of treatment the HAMA decrease values in two groups were less than those before the treatment (P<0.01). There was on significant difference in response rate between the2groups. There was significant difference in TESS between the2groups.
4.Conclusion:ASF is one of the effective therapies with less emergent symptom in treating GAD, which is worth studying and spreading further more.
chapter2. The pharmacodynamics research on the treatment of anxiolytic with ASF
1.Objective:using elevated plus-maze as a measure of anxiety in the rat to observe anxiolytic effect of ASF particles and do preliminary study of its mechanism.
2.Method:Sixty male wistar rats were randomly divided into6groups (10rats/group):normal control group, model group, diazepam group, and high,medium and low dose ASF group; then used chronic emotional stress method to establish anxiety rat model and gave the drug intervention at the same time.40g.Kg-1for the high dose group.20g.Kg-1for the middle dose group,and10g.Kg-1for the low dose group and the saline for the control group;meanwhile,made the insomnia model for15day.Then evaluate their behavior by adopting internationally recognized elevated plus-maze test (EPM)
3.Anxiety rats model preparation:using chronic emotional stress method to establish anxiety rat model by elevated plus-maze.
4. Used statistical software SPSS13.0to detail the data.During the multi-groups compares uses the single factor variance analysis (One-Way ANOVA) to examine; All statistical analysis has the significance boundary as P<0.05.
5. Result:Compared with normal control group, percentage rate and frequency rate of model group into open arm time was significantly reduced, and that of rats from diazepam group and ASF dose high, medium and low dose group was significantly raised comparing with model group, the most obvious raise group was ASF medium dose group.
6. Conclusion:The pharmacodynamics research on the treatment of anxiolytic with ASF shoud that ASF provides anti-anxiety effect by elevated plus-maze(EPM) as a measure of anxiety
Chapter3. The laboratory explore on the treatment of anxiolytic with Anshenfang
1.Objective:To observe anxiolytic effect of ASF particles and do preliminary study of its mechanism.
2.Method:①. Sixty male wistar rats were randomly divided into6groups (10rats/group):normal control group, model group, diazepam group, and high,medium and low dose ASF group;②.Using chronic emotional stress method to establish anxiety rat model and gave the drug intervention at the same time to evaluate their behavior by adopting internationally recognized elevated plus-maze test (EPM)
③.After behavioral observation, anesthesias executed rats of normal control group, model group, ASF medium dose group; separated rats hippocampus on the ice stage.
④.Separated clarified liquid to determine the content of glutamic (GLU) and y-aminobutyric acid (GABA) by using western blot method;
⑤.Evaluated GABAA receptor, N-methyl-D-aspartate receptor1expression through immunohistochemistry.
⑥. Detect the receptor gene by the fluorescence quantitative polymerase chain reaction(FQ-PCR).
3.Used statistical software SPSS13.0to detail the data.During the multi-groups compares uses the single factor variance analysis (One-Way ANOVA) to examine; All statistical analysis has the significance boundary as P<0.05.
4.ResuIt:Compared with normal control group, model of glutamate in hippocampal tissue expression increased significantly compared with the normal control group and γ-aminobutyric acid expression decreased obviously; compared with the model group, glutamate acid expression in diazepam and medium ASF dose group was significantly low and y-aminobutyric acid expression was increased. While, there was no significant difference when comparing ASF medium dose group with diazepam group; compared with control group, GABAA receptor immune positive cells expression in model group was lower and amount of Immune positive cells expression of N-methyl-D-aspartate receptor1was increased; compared diazepam dosage group and ASF medium dose group with model group, the N-methyl-D-aspartate receptor1expression of the former were significantly reduced and GABAA receptor expression level were increased; while there was no significant difference in ASF medium dose group and diazepam group.
5.Conclusion:ASF provides anti-anxiety effect,①.increase the content of y-aminobutyric acid in the brain, strengthen GABAA receptor and the receptor gene expression,reduce glutamic acid content,and decrease N-methyl-D-aspartate receptor1and the receptor gene expression. The best one is ASF medium dose group.
焦虑是精神障碍五大症状群中常见的一种,焦虑障碍作为神经症的一个独立的亚型,其发病率正呈现逐年上升趋势。焦虑障碍又称焦虑症,其主要有两种临床形式--广泛性焦虑障碍和惊恐障碍。广泛性焦虑障碍(generalized anxiety disorder, GAD)又称慢性焦虑症,其主要表现为对实际并不存在的威胁或危险以及身体健康过分的担心、紧张和害怕,并伴有口干、胃肠不适、恶心、腹泻、呼吸困难、心悸、尿频尿急、男性阳痿、女性月经不调、出汗、面色潮红及肌肉紧张、手颤、头痛、坐立不安等症状,另外还伴有注意力不集中、记忆力减退、睡眠障碍等症状。
随着我国经济发展和社会文明的进步,人们早已消除了对精神疾病的狭隘误解,正确的认识使得人们对自我心理健康的关注程度有了较大提高,因觉察到自身焦虑症状而主动就医的人数越来越多,焦虑障碍已经成为综合医院门诊、心理咨询和治疗机构中常见的一类疾病。越来越多患有焦虑症状的人群希望能够得到及时、有效的诊断和治疗,为此,全球各国医务工作者正在积极开展对焦虑障碍发病机制以及预防治疗方案等方面的研究。
目前,现代医学在对焦虑障碍的发病机制研究方面已经取得了很大进展,药物治疗方面也已具备了较为完善的理论。临床治疗多采用以地西泮为代表的苯二氮卓类药物和丁螺环酮等非苯二氮卓类药物,前者在使用初期起效很快但常伴有嗜睡、便秘、肌松、记忆力减弱等副作用,后者虽副作用较少,但起效通常较慢,另外该类药物由于容易形成依赖和耐受以及戒断反应等因素,故也在一定程度上限制了应用。因此,寻找疗效确切、副作用小的抗焦虑药物就成为了目前抗焦虑研究领域的重要课题。
我们的课题研究首先通过收集查阅大量相关文献,在学习借鉴现代医学对焦虑障碍的研究现状之后,找到了焦虑障碍与中医学情志病之间的相关性和共同点,并在总结了中医学历代、各学派学者对情志病的调治方法的基础之上,在博士生导师、知名中医陈宝田教授指导下,确立了清热除焦、调和营卫、安神定志的治疗原则,制定了安神方(ASF)的组方用药。教授结合个人近50年临床经验,在安神方中配伍运用柴胡、酸枣仁、桂枝、龙骨、牡蛎等药物,用于治疗焦虑症见焦躁易怒、胸胁胀闷、惊悸不安、失眠健忘、肢体困乏、口苦咽干、舌红脉弦数的患者。此次研究通过观察安神方治疗广泛性焦虑障碍(GAD)的临床疗效、并进一步通过动物试验探讨安神方治疗焦虑障碍的药效学及相关作用机制。旨在充分发挥祖国医学整体观念、辨证论治这些优势特点的同时,结合现代医学的实验方法,更好的解释其作用效果及机理,为更好的治疗焦虑障碍探索铺就出一条有效之路。
一、安神方治疗广泛性焦虑障碍(GAD)的疗效观察
1.目的：通过临床观察探讨安神方治疗广泛性焦虑障碍(GAD)的临床效果,观察症侯及临床整体疗效以及收集整理相关检查所得的结果,客观的评价安神方治疗GAD的临床疗效。
2.方法：
2.1病例与分组：将符合纳入标准的120例GAD入组患者随机分为治疗组和对照组,每组60例；
2.2用药：治疗组口服“安神方”(由南方医院中药房提供,方剂组成为：柴胡25g,桂枝10g,黄芩10g,党参15g,白芍15g,生龙骨30g,生牡蛎30g,炒枣仁15g,远志10g,夜交藤30g,甘草10g等),每日1剂,分2次,晨起空腹及晚睡前30min各服一次；对照组口服地西泮(由天津利生制药股份有限公司提供,2.5g/片,100片/瓶)口服,1片/次,晨起空腹及晚睡前30min各服一次。两组均以连续服药4周为1个疗程。治疗期间不同用其他抗焦虑及镇静安眠药。两组均于诊疗中同时注意对患者进行必要的心理疏导,避免情绪波动。
2.3改变指标：分别于治疗前及治疗后1、2、4周末各进行一次汉密尔顿焦虑量表(HAMA)评定。以HAMA减分率(T)作为疗效评定指标, T=(治疗前总分-治疗后总分)/治疗前总分×100%。T小于25%为无效,25%～49%为好转,50%～75%为显著好转,大于75%为临床痊愈；总有效率=(痊愈+显著+好转)/总例数×100%。
2.4临床整体疗效、不良反应及安全性判定：以汉密尔顿焦虑量表(HAMA)和副反应量表(TESS)评价疗效和不良反应及其程度。并于治疗前后检查：体温,心率,血压,呼吸；血、尿、便常规；肝、肾功能；心电图等项目,以此对两组服药后的不良反应以及药物的安全性进行全面的评价。
2.5统计学方法：采用统计软件SPSS13.0进行统计学处理和分析。试验数据计量资料采用均数加减标准差(X±S)表示。治疗前后剂量资料比较采用配对样本t检验；两组间计量资料比较采用独立样本t检验；治疗后组间症候疗效比较采用协方差分析；等级资料分布情况采用秩和检验及非参数检验。计数资料以百分率表示,采用卡方检验。所有统计数据均以P<0.05作为有显著性差异的界限。
3.结果：治疗4周后,两组HAMA分值均较治疗前有显著差异(P均<0.01),两组间比较差异无统计学意义(P>0.05)。治疗组与对照组TESS量表评分比较差异有统计学意义(P<0.05)。
4.结论：初步确认了安神方治疗GAD安全、有效且不良反应较小,值得进一步研究推广。
二、安神方抗焦虑作用的药效学研究
1.目的：选用大鼠高架十字迷宫试验模型(EPM),观察安神方的抗焦虑作用。
2.方法：将60只健康雄性Wistar大鼠随机分为6组(10只/组)：正常对照组、模型组、地西泮组(DZP)和安神方(ASF)高、中、低剂量组。采用慢性情绪应激的方法建立焦虑大鼠模型,同时给予药物干预,以国际公认的高架十字迷宫试验对其进行药效行为学评价。正常对照组不接受任何处理,自由饮水和摄食。其余各组定时喂水训练7天,之后开始应激试验,在上述两个时间段内给予不确定空瓶刺激,维持1天1次或2次,持续2周。在不确定空瓶刺激期间,地西泮组给予1mg/kg灌胃,模型组每天给予等容积生理盐水,ASF高、中、低剂量组每天剂量分别为：40g/kg、20g/kg、10g/kg。于第21天,ASF组末次给药2h、模型组及DZP组末次给药0.5h后,进行高架十字迷宫行为学测定,大鼠均提前2h进入测试实验室适应环境。
3.仪器：大鼠高架十字迷宫(EPM)采用国际通用的方法制作。
4.统计学方法：试验数据采用SPSS13.0统计软件进行统计分析,组间比较采用单因素方差分析,多重比较采用SNK法。试验数据均以均数加减标准差(x±s)表示,P<0.05为有统计学意义。
5.结果：与正常对照组比较,模型组大鼠进入开臂的时间百分率(OT%)次数百分率(OE%)均明显减少；DZP组与ASF高、中、低剂量组大鼠进入开臂的时间百分率(OT%)、次数百分率(OE%)均较模型组显著上升,其中以ASF中剂量组上升最明显。
6.结论：行为学试验结果显示：DZP组、ASF高、中、低剂量组同模型组比较, EPM中大鼠OE%和OT%值均升高(P<0.05),尤其是ASF中剂量组升高最明显(P<0.01),其作用效果与DZP相当,这进一步证实了ASF对大鼠焦虑状态的干预作用,而且以中剂量给药干预效果最好。
三、安神方抗焦虑作用的形态学及相关机制研究
1.目的：观察安神方(ASF)的抗焦虑作用,并初步研究其机制。
2.方法：①.将所取雄性Wistar大鼠随机平均分为6组：正常对照组、模型组、DZP组和ASF高、中、低剂量组；
②.采用慢性情绪应激的方法建立焦虑大鼠模型,同时给予药物干预,以国际公认的高架十字迷宫模型(EPM)试验对其行为学进行评价；
③.行为学观察后,将正常对照组、模型组、DZP组及ASF高、中、低剂量组大鼠立即麻醉处死、断头,于冰台上剥离大鼠的海马组织。
④.在光学显微镜下采用HE染色法观察大鼠脑组织内神经元形态结构的改变和胶质细胞的增生情况；用尼氏染色法观察神经元受损和尼氏体的丢失情况。
⑤.用免疫组化法观察ASF对焦虑大鼠脑组织内氨基酸类神经递质--Y-氨基丁酸(GABA)和谷氨酸(Glu)含量的影响；
⑥.用Western bolt法观察ASF对焦虑大鼠脑组织内氨基酸类神经递质受体-GABAA 受体(GABAARα1)和Glu受体(N-甲基-D-天冬氨酸受体1即NMDAR1)表达的影响；
⑦.用荧光定量PCR法观察ASF对焦虑大鼠脑组织内氨基酸类神经递质受体-GABAA受体(GABAARα1)和Glu受体(N-甲基-D-天冬氨酸受体1即NMDAR1)基因表达的影响。
3.仪器：MilliQS纯水／超纯水系统、6219型PH计、BCD-196DT冰箱、CR703恒温振荡器、SFG-02B电热恒温干燥箱、TS-1脱色摇床、Eoipse Ti-s倒置显微镜、MI-22FOEGC微波炉、RF2000荧光检测器、PHS-2C精密酸度计、Avanti30冷冻离心机、BA110S电子天平、TiThermocyclerRea-timepCR仪、CA-92-2垂直层流洁净工作台、DYCP水平电泳槽、DYY6B型稳压稳流电泳仪、sigma3K30超速冷冻离心机、MX3005P荧光定量PCR仪及分析软件MXPro4.01
4.试剂：PBS缓冲液(ph7.2—7.4)、高效切片石蜡、中性甲醛、无水酒精、二甲苯、浓盐酸、苏木精、伊红染、中性树脂、50ml离心管、普通玻片、盖玻片、玻片盒、玻片架、谷氨酸(Glu)购自美国SIGMA,公司批号分别为43907190；Y-氨基丁酸(GABA)购自美国SIGMA公司,公司批号116K5005；乙腈购自美国TEDIA公司；GABAARal兔IgG多克隆抗体购自美国santa curz公司；N-甲基-D-天冬氨酸受体KNMDAR1)兔IgG多克隆抗体购自美国santa curz公司;2mlPCR荧光管购自Axgen公司；Nalater:AMBION公司；2000bpDNALadder:广州瑞真生物科技有限公司；焦碳酸二乙醋(DEPC) treated water:大连TaKaRa宝生物公司；Trizol:Invitrogen公司、RNA反转录试剂盒：Takara公司、SyBrI荧光定量试剂盒购自Gene copoeia公司、琼脂糖购自Sigma公司,其余试剂均为进口及国产分装试剂。
5.统计学方法：试验数据采用SPSS13.0统计软件进行统计分析,组间比较采用单因素方差分析,多重比较采用SNK法。试验数据均以均数加减标准差(x±s)表示,以P<0.05为有统计学意义。
6.结果：
①.正常对照组：海马CA3区细胞排列整齐,细胞核圆而大,核仁清晰可见,锥体细胞排列整齐,层次紧密,尼氏体深染；模型组：海马CA3区锥体细胞排列紊乱,神经细胞出现明显的病理变化,染色质溶解,部分细胞脱失,胞体固缩呈多角形或不规则形,胞质浓缩核膜界限不清,锥体细胞排列松散,细胞缺失,尼氏体浅染甚至溶解；DZP组：海马CA3区锥体细胞排列比较整齐,少部分神经细胞出现病理变化,少部分细胞染色质溶解,少部分胞体固缩呈不规则形,锥体细胞排列部分松散,锥体细胞与中剂量组相当,细胞部分缺失；ASF低、中、高剂量组：海马CA3区锥体细胞排列比较整齐,神经细胞部分出现病理变化,部分细胞染色质溶解,部分细胞脱失,胞体固缩呈不规则形。其中中剂量组细胞形态修复最好,ASF低剂量组锥体细胞排列比较整齐,比模型组锥体细胞明显增多,细胞部分缺失,尼氏体浅染；ASF中剂量组锥体细胞排列比较整齐,锥体细胞与高剂量组相当,细胞部分缺失,尼氏体染色较高剂量组深染,与DZP剂量组相当；ASF高剂量组锥体细胞排列比较整齐,比低剂量锥体细胞有所增多,细胞部分缺失,尼氏体浅染。
②.模型组大鼠海马组织中Glu的表达量较正常对照组显著上升,GABA的表达量明显下降;地西泮组和安神方中剂量组同模型组比较,大鼠海马组织中Glu的表达量显著降低和GABA的表达量明显提高,安神方中剂量组与地西泮组比较差异不显著；
③.模型组与正常对照组比较,GABAA受体表达减少,Glu受体表达增多；地西泮组和安神方高、中、低剂量组分别同模型组进行比较,Glu受体的表达水平均降低,GABAA受体的表达水平均提高；安神方中剂量组与地西泮组比较无明显差异。
④.模型组与正常对照组比较,GABAA受体基因减少,Glu受体基因增多；地西泮组和安神方高、中、低剂量组分别同模型组进行比较,Glu受体基因水平均降低,GABAA受体受体基因水平均提高；安神方中剂量组与地西泮组比较无明显差异。
7.结论：安神方具有抗焦虑的作用,具体表现在：
①可以修复焦虑模型大鼠中枢神经系统中海马区出现的病理性变化；
②可以提高大鼠中枢神经系统中GABA的含量,降低Glu的含量；
③可以增加大鼠中枢神经系统中GABA受体的表达,减少Glu受体的表达；
④可以增加大鼠中枢神经系统中GABA受体基因的表达,减少Glu受体基因的表达。
以上研究结果进一步证实了安神方对大鼠焦虑状态的干预作用,而且以中剂量给药干预效果最好。
While advances in science and technology have brought huge amounts of benefits to modern societies, they have also caused numerous side effects that increasingly threaten people's heath. The modern world is fast-paced and filled with fierce competition at a global level, and as a result, millions of people are under more pressure than ever before, and most would have to face deteriorated working conditions such as longer working hours, the requirement of higher level of concentration, insufficient physical exercises, etc.. World Health Organization (WHO) reported that nowadays people pay more and more attention to mental health and psychological welbeing [REF]. It also pointed out that among the10diseases that are most costly to taxpayers,4are classified as mental illness. In addition, the treatment of mental diseases costs a quarter of the total budget on health. These facts have shown that mental health has brought an increasing amount of attention, not only from medical practitioners, but also governments and their citizens.
Anxiety disorder is one of the biggest type of mental illness and it shows an increasing incidence in recent years. Anxiety disorder covers several different forms of abnormal and pathological fear and anxiety. It is divided into Generalized Anxiety Disorder (GAD) and Panic Disorder (PD), where GAD is a common chronic disorder characterized by long-lasting anxiety that is not focused on any one object or situation. Those suffering from GAD are frequently accompanied by physiological symptoms such as dry mouth, upset stomach, nausea,diarrhea, dyspnea, palpitations, frequent urination, impotentia,menstrual disorder, sweating, flushed face,①muscle tension and spasms.headache, short attention span, hypomnesis and sleep disorder. In China, along with the economic development in recent years, people have realized the importance of mental health. As people become more active in seeking medical advice on symptoms of anxiety disorder, hospitals and specialized clinics have admitted an increasing number of patients with such disease, which has driven a surge in research in the physiology of anxiety disorder and its treatment.
Currently, modern medicine not only has made great progress in the pathogenesis of anxiety disorders, but also has a sound theoretical in drug treatment. There are two classes of drugs being mostly used in clinical treatment, benzodiazepine drugs such as Diazepam and non-benzodiazepine drugs such as Buspirone. The former ones effect quickly in the beginning, but often accompanied by lethargy, constipation, muscle relaxants, memory weakened, and other side effects. The latter ones, with fewer side effects, effect slower. Moreover due to the easy formation of dependence and tolerance, and withdrawal reactions, to some extent limits the application of this classes of drugs. Therefore to find out the drugs with efficacy and fewer side effects, has become an important topic of the current anti-anxiety research areas.
After accessing large amounts of literatures and leaning from anxiety disorders researches of modern medicine, our subject of study finds out correlations between anxiety disorders and these disorders of Chinese medicine. Under the guidance of the doctoral mentor, the renowned Chinese Medicine Professor Chen Baotian, on the basis of summarizing,we establish three treatment principles, and set up AnShenfang combined with almost50years of clinical experiences of professor Chen, we use chaihu,guizhi,longgu,muli and others medicines in AnShenfang for the treatment of anxiety. This study observes the clinical efficacy of AnShenfang in treatment of Generalized Anxiety Disorder (GAD), and further with animal experiments, investigates the pharmacodynamics, morphology and the mechanism of actions of AnShenfang. Aimed to give full play to the concepts of Chinese Medicine and the advantages of treatment based on syndrome differentiation,combined with modern medical experiments, our study is designed to better explained the role of effects and mechanisms of AnShenfang, in order to explore and pave out an effective way for better treatment of anxiety disorders.
chapter1. Observation of AnShenFang in Treating Generalized Anxiety Disorder
l.Objective:To investigate the efficacy of AnShenFang(ASF) for patients with general anxiety disorder (GAD). To objectively estimate the therapical effect of "AnShenfang" in treating GAD through the clinical overall curative effect as well as the finding.
2.Methods:Clinical curative effect observation:120cases according with the diagnostic standard were equally divided into (ASF) treatment group and western medicine treatment(DZP) control group at random.
During treatment:The patients were entered a4-week randomized contralled triad.60patients of control group were given DZP and other60of treatment group were given ASF.other sedative and anxiolytic medicines should be abstinent.Mental nursing is necessary besides the therapy of during treatment
The HAMA were measured before and after3,6,12months of the treatment. The results were compared with ASF treatment group and western medicine treatment control group.
Assess the benefits and adverse reaction of ASF and DZP in the treatment of GAD, according to the level of Hamilton Anxiety Scale (HAMA) and Treatment Emergent Symptom Scale (TESS)Before and after treatment, temperature, heart rate, blood pressure, renal and hepatic funtion, plasmal electrolyte and electrocardiogram were assessed and compared.
3. Results:After4weeks of treatment the HAMA decrease values in two groups were less than those before the treatment (P<0.01). There was on significant difference in response rate between the2groups. There was significant difference in TESS between the2groups.
4.Conclusion:ASF is one of the effective therapies with less emergent symptom in treating GAD, which is worth studying and spreading further more.
chapter2. The pharmacodynamics research on the treatment of anxiolytic with ASF
1.Objective:using elevated plus-maze as a measure of anxiety in the rat to observe anxiolytic effect of ASF particles and do preliminary study of its mechanism.
2.Method:Sixty male wistar rats were randomly divided into6groups (10rats/group):normal control group, model group, diazepam group, and high,medium and low dose ASF group; then used chronic emotional stress method to establish anxiety rat model and gave the drug intervention at the same time.40g.Kg-1for the high dose group.20g.Kg-1for the middle dose group,and10g.Kg-1for the low dose group and the saline for the control group;meanwhile,made the insomnia model for15day.Then evaluate their behavior by adopting internationally recognized elevated plus-maze test (EPM)
3.Anxiety rats model preparation:using chronic emotional stress method to establish anxiety rat model by elevated plus-maze.
4. Used statistical software SPSS13.0to detail the data.During the multi-groups compares uses the single factor variance analysis (One-Way ANOVA) to examine; All statistical analysis has the significance boundary as P<0.05.
5. Result:Compared with normal control group, percentage rate and frequency rate of model group into open arm time was significantly reduced, and that of rats from diazepam group and ASF dose high, medium and low dose group was significantly raised comparing with model group, the most obvious raise group was ASF medium dose group.
6. Conclusion:The pharmacodynamics research on the treatment of anxiolytic with ASF shoud that ASF provides anti-anxiety effect by elevated plus-maze(EPM) as a measure of anxiety
Chapter3. The laboratory explore on the treatment of anxiolytic with Anshenfang
1.Objective:To observe anxiolytic effect of ASF particles and do preliminary study of its mechanism.
2.Method:①. Sixty male wistar rats were randomly divided into6groups (10rats/group):normal control group, model group, diazepam group, and high,medium and low dose ASF group;②.Using chronic emotional stress method to establish anxiety rat model and gave the drug intervention at the same time to evaluate their behavior by adopting internationally recognized elevated plus-maze test (EPM)
③.After behavioral observation, anesthesias executed rats of normal control group, model group, ASF medium dose group; separated rats hippocampus on the ice stage.
④.Separated clarified liquid to determine the content of glutamic (GLU) and y-aminobutyric acid (GABA) by using western blot method;
⑤.Evaluated GABAA receptor, N-methyl-D-aspartate receptor1expression through immunohistochemistry.
⑥. Detect the receptor gene by the fluorescence quantitative polymerase chain reaction(FQ-PCR).
3.Used statistical software SPSS13.0to detail the data.During the multi-groups compares uses the single factor variance analysis (One-Way ANOVA) to examine; All statistical analysis has the significance boundary as P<0.05.
4.ResuIt:Compared with normal control group, model of glutamate in hippocampal tissue expression increased significantly compared with the normal control group and γ-aminobutyric acid expression decreased obviously; compared with the model group, glutamate acid expression in diazepam and medium ASF dose group was significantly low and y-aminobutyric acid expression was increased. While, there was no significant difference when comparing ASF medium dose group with diazepam group; compared with control group, GABAA receptor immune positive cells expression in model group was lower and amount of Immune positive cells expression of N-methyl-D-aspartate receptor1was increased; compared diazepam dosage group and ASF medium dose group with model group, the N-methyl-D-aspartate receptor1expression of the former were significantly reduced and GABAA receptor expression level were increased; while there was no significant difference in ASF medium dose group and diazepam group.
5.Conclusion:ASF provides anti-anxiety effect,①.increase the content of y-aminobutyric acid in the brain, strengthen GABAA receptor and the receptor gene expression,reduce glutamic acid content,and decrease N-methyl-D-aspartate receptor1and the receptor gene expression. The best one is ASF medium dose group.
引文
[1]沈渔邮主编.精神病学[M].第4版.北京：人民卫生出版社,2003.
[2]萨默斯·朗文当代英语大辞典[M].北京：商务印书馆,2004：60.
[3]张奇文.英汉双解精选医学词典[M].北京：人民卫生出版社,1990：465.
[4]American Psychiatric Association.Diagnostic and Ststistical Manual of Mental disorders.4th ed.Washington DC:APA 1994
[5]Murray CJ, Lopez AD.Altemative Projeetions of mortality and disability by cause 1990～2020.Global Burden of Disease Study [J] J.Lancet,1997,349: 1498-1504
[6]Ustun TB, Sartourius N. Mental illness in general healthcare. New York: Wiley,1995.323-375
[7]Kessler RC, Berglund P, Demler O.Lifetime prevalence and age-ofonset distributions of DSM-IV disorders in the National Comorbidity Survey Replication. Arch Gen Psychiatry 2005,62:593-602
[8]Pigott TA.Gender differences in the epidemiology and treatment of anxiety disorders.J Clin Psychiatry,1999,60(Supp118):4
[9]袁勇贵.焦虑障碍共病研究.[J].中国临床康复,2004,8(33)：7544-7547
[10]陈复平.神经症的流行病学调查[J].中华神经精神科杂志,1986,19(3)：301
[11]区丽明, 叶瑞繁, 陈剑等.广州地区综合医院门诊抑郁/焦虑障碍流行病学调查[J].中国临床心理学杂志,2009,17(1)：61
[12]向孟泽,单友荷,段诚凤,等12地区神经症流行病学调查[J].中华神经精神科杂志,1986,19(2)：87
[13]王欣,王守勇.酸枣仁汤临床与实验研究评述[J].中成药,2005,25(5)：414
[14]工泽帅,马建军,李学,等.神经内科门诊患者焦虑和抑郁症状的调查研究[J].中国实用神经疾病杂志,2009,12(1)：51-53
[15]Alonso J, Angermeyer MC, Bernert S, et al. Prevalence of mental disorders in Europe:results from the European Study of the Epidemiology of Mental Disorders (ESEMeD) project. Acta Psychiatr Scand 2004,109(suppl 420): 21-27
[16]Wittchen H-U, Nelson CB, Lachner G. Prevalence of mental disorders and psychosocial impairments in adolescents and young adults.Psychol Med 1998,28:109-126
[17]Murray CJ, Lopez AD.Alternative Projections of mortality and disability by cause 1990-2020.Global Burden of Disease Study [JJ.J.Lancet,1997,349: 1498-1504
[18]Leckman JF,Merikangas KR,Pauls DL,et al.Anxiety disorders and depression:contradictions between family study data and DSM-Ⅲ conventions.Am J Psych Iatry,1983,140(7):880
[19]Kendler KSJ。Neale MC,Kessler RC,eral.Majorde Pression and generalized anxiety disorder:Same genes,(Party)different environment[J].Arch Gen Psychiatry,1992,49:716-722
[20]Roy MA.Neale MC,Pedersen NL,etal.Atwinstudy of generalized anxiety disorder and major depression[J].Psyehol Med,1995,25:1037-1049
[21]Kendle KS, Walters EE.Neale MC.et al.The structure of the genetic and environment risk factors six major psychiatric disorders in women:Phobia, GAD.Panic disorder, bulimia major depression and aleoholism[J]. Areh Gen Psyehiatry,1995,52:374-383
[22]马震祥,刘玉局.广一泛性焦虑遗传效应和遗传方式研究[J].临床精神医学杂志,2006,16(6)～337
[23]Wu JC, Buehsbaum MS,Hershey T,et al.Positron emission tomography study of generalized anxiety disorder[J].Biol Psychiatry,1991,29:1181-1199
[24]万黎,刘建,吴文源,等.焦虑症患者平静及紧张状态局部脑血流灌注改变[J].中华核医学杂志,2002,22：206-208
[25]孙延强,赵相英,郭秀珍,等.焦虑症、抑郁症患者与正常健康人颅脑CT对比研究[J].中国行为医学科学,2005,12(14)：1082-1083
[26]王维治主编.神经病学.第5版.人民卫生出版社,2006：184-185
[27]Gerra G,Zaimovic A,Zambelli U,et al.Neuroendcrine responses to psychological stress in adolescents with anxiety disorder. Neuropsychobiology,2000,42:82
[28]Guimaraes FS, Carobrez AP, De Agniar JC, et al.Anxioly effect in the elevated plus-maze of the NMDA receptor antagonist AP7 mieroinjeeted into the dorsal periaqueduetalgrey[J].Psychopharmaeology(Beri),1991,103(1):91-94
[29]Matheus MG, Guimaraes FS.Antagonism of non-NMDA receptors in the dorsal periaqueduetal grey induces anxiolytic effeet in the elevated plus maze[J].Psychophamaeology(Berl),1997,132(1):14-18
[30]李东亮,张朝.基础神经生物学[M].北京：人民军医出版社,2006,122
[31]Sibile E,Pavlides C,Benke D, Toth M.Genetic inactivation of the Serotonin(lA) receptor alpha subuzlits,reduetion of GABA(A) receptor binding,and belzodiazePine-resistant anxiety[J].JNeurosci,2000,20:2758-2765
[32]Nutt DJ.Neurobiologieal mechanism in generealized anxiety disorder [J].J Clin Psychiatry,2001,62(Suppl):22-27
[33]Ballenger JC.Overview of different pharmaeotherapies for attaining remission in generalized anxiety disorder[J].J Clin Psyhiatry,2001,62(Supp119):11-19
[34]张瑞华,李丽琴,王惠芳,等.A型氨基丁酸受体结构及其有关药物[J].生命的化学,2003,23(6)：441-444
[35]Serra M, Pisu MG,Littera M, et al.isolation induced decreased in both the abundanee of neuroaetive steroids and GABAA receptor function in rat brain[J].J Neuroehem,2000,75:732-740
[36]Kathryn M, Connor, Jonathan R,et al.Genralized anxiety disorder:neurobiologieal and pharmaeotherapeutic persectives[J].Biological Psychiatry,1998,44(12):1286-1294
[37]Hoyer D,Clarke DE,Fozard JR,et al.1.International union of pharmaeoloy classification of receptors for 5-hydroxytryptamine(serotonin)[J].Pharmacol Rev,1994,46:157-203
[38]吴东辉,刘少文.广泛性焦虑障碍的神经生物学机制[J].国外医学·精神病学分册,2003,30(2)：124-126
[39]工振,肖泽萍,陈迁,等.焦虑症的生化病理机制探讨[J].临床精神医学杂志,2003,13(1)：1～3
[40]Baldwin D, Rudge S.The role of serotonin in depression and anxiety[J].Int ClinPsychoPharmacol,1995,9(suppl4):41-45
[41]谢健,高力舒.焦虑症艾森克个性问卷与血浆去甲肾上腺素、总皮质醇及白介素-2测定及相关分析[J].江西医药,2006,41(5)：266-268
[42]Kathryn M, Connor, Jonathan R.T.et al.Generalized anxiety disorden:neurobiological and Pharmaeotherapeutic perspectives[J].Biol Psychiatry,1998,44(12):1286-1294
[43]中华医学会精神科分会CCMD-3中国精神障碍分类与诊断标准[M].第3版.济南：山东科学技术出版社,2001,106～107
[44]王焕林.临床精神医学[M].北京：人民军医出版社,2003,324
[45]张德昌.医学药理学[M].北京：北京医科大学、中国协和医科大学联合出版社,1998.431.
[46]梁正英等.丁螺环酮在社交焦虑障碍治疗中的应用.贵阳医学院学报,2005(5)：445-446.
[47]翟金国等,曲唑酮与丁螺环酮治疗广泛性焦虑障碍的对照研究.中国新药与临床杂志,2008(11)：832-835
[48]邹万芹.新型抗抑郁药的临床应用及研究进展[J].中国药房,2008,19(14)： 1105.
[49]白秋江,郑敏.选择性5-羟色胺再摄取抑制剂致不良反应文献分析[J].中国药房,2007,18(20)：1583.
[50]孙松涛.柴胡加龙骨牡蛎汤加减治疗广泛性焦虑症72例[J].中医杂志,2000,41(2)：95.
[51]雷翔,柴胡加龙骨牡蛎汤临证举隅[J].中国民族民间医药,2010(9)：138.
[52]倪良玉,柴胡加龙骨牡蛎汤的临床应用体会[J].山西中医,2005(2)：58-59
[53]王守勇,谢鸣、王欣,酸枣仁汤组分与抗焦虑效用的相关性研究[J].中国实验方剂学杂志,2010(3).
[54]王业民、周建军、孔令东,半夏厚朴汤及其组成中药抗焦虑和抑郁研究进展[J].世界科学技术,2003(4).
[55]马行,库宝善,徐英,等.复方郁金胶囊抗抑郁抗焦虑的作用[J].中国临床康复,2006,10(15)：56.
[56]徐志伟,王文竹,苏俊芳,等.逍遥散和丹栀逍遥散抗焦虑作用的实验研究[J].广州中医药大学学报,2006,23(4)：330.
[57]贺玉琢.以群居干扰实验探讨加味逍遥散的抗焦虑作用及其活性成分(I)[J].国外医学-中医中药分册,2004,26(4)：250.
[58]加减逍遥散与氯硝西泮治疗焦虑症状群的对照观察[J].海峡药学,2004,(05).
[59]王玲姝,郑祖艳.针药结合对焦虑性神经症患者5-羟色胺及去甲肾上腺素 的影响[J].针灸临床杂志,2009,25(7)：13-14.
[60]罗文政,刘海静,梅尚英,等.靳三针治疗广泛性焦虑症的临床观察[J].中国中西医结合杂志,2007,27(3)：201-203.
[61]王贯民,针刺治疗广泛性焦虑症21例[J].河南中医,2007,27(7),55-56
[62]张涛,邵沛,陈妙.针药结合治疗焦虑症35例临床观察[J].针灸临床杂志,2006,22(12)：13-14.
[63]赵天辉,经络氧疗法治疗焦虑障碍60例疗效观察[J].新中医2006,38(2)：66-67.
[64]霍小宁,耳压法治疗广泛性焦虑28例临床观察[J].甘肃中医,2005,18(4)：30.
[65]太鑫,针灸结合心理疗法治疗广泛性焦虑障碍的临床观察[J].中医药信息,2010,27(2)：72-73.
[66]沈莉,颜红,冯辉,针药结合治疗广泛性焦虑症临床观察[J].上海针灸杂志,2007,26(3)：3-4
[67]黄奕,赵长鹰.浅述失眠症的心理治疗方法[J].陕西中医,2005,26(5)：479
[68]胡师焕.《内经》中有关心理学的理论观点[J].湖北中医学院学报,2004,6(3)：24
[1]中华医学会精神科分会.中国精神障碍分类与诊断标准[M].3版.济南：山东科学技术出版社,2001：106-107.
[2]张明园.精神科评定量表手册[M].长沙：湖南科学技术出版社,2003.
[3]中药新药临床研究指导原则(试行)[S].北京,中国医药科技出版社.2002.
[4]Kessler RC, Berglund P, Demler O.Lifetime prevalence and age-ofonset distributions of DSM-Ⅳ disorders in the National Comorbidity Survey Replication. Arch Gen Psychiatry 2005,62:593-602.
[5]熊曼琪.伤寒论[M].北京：中国中医药出版社,2007：202.
[6]吴斌龙.桂枝汤方证发微[J].辽宁中医药大学学报,2011,13(2)：99-100.
[7]贺一新,赵素霞,崔瑛.酸枣仁抗焦虑活性物质分析[J].中药 材,2010,33(2)：229-231.
[8]傅晶,张东明,陈若芸.远志属植物的皂苷类成分及其药理作用研究进展[J].中草药,2006,37(1)：144-146.
[9]Li X,Matsumoto K,Murakami Y,et al.europrotective effects of Polygonum multiflorum on nigroatriatal dopaminergic degeneration induced by paraquat and maneb in mice[J].Pharmacol Biochem Behav,2005,82(2):345-352.
[10]王维勋,孙付军,李芳,张希林,张军.柴胡加龙骨牡蛎汤对焦虑模型大鼠单胺类递质的影响[J].中药新药与临床药理,2008,19(5)：340-342.
[1]Pellow S, Chopin P, File SE, et al. Validation of open-closed arm entries in an elevated p lus-maze as a measure of anxiety in the rat[J]. Journal of Neuroscience Methods,1985,14 (3):149-167
[2]Montgomery S.Persistent c-fos expression in the brains of mice with soeial stress[J].Neurosei-Res,1996,26(2):157
[3]Holmes A,Rodgers R.J.Responses of Swiss-Webster mice to repeated plus-maze experience:further evidence for a qualitative shift in emotional state.Pharmacology Biochemistry and Behavior,1998,60(2):473-488
[4]Gard PR,Haigh SJ,Cambursano PT,et al.Strain differences in anxiolytic effects of losartan in the mouse.Pharmacology Biochemistry and Behavior,2001,69(1-2): 35-40
[5]Pellow S,Chopin P,File SE,et al.Validation of open:closed arm entries in an elevated plus-maze as a measure of anxiety in the rat[J].J Neurosci Methods,1985,14(3):149-167
[6]Lister RG.The use of a plus-maze to measure anxiety int the miee [J].Psycho Pharmaeology (Berl),1987,92(2):180-185
[7]Lister RG.The use of a plus-maze to measure anxiety in the mice [J].Psvchophaimacolony(Berl),1987,92(2):180-185
[8]程九清,李勇辉,隋南.基于啮齿动物的决策行为研究及其脑机制[J].心理科学杂志,2008,16(5)：721—725
[9]徐叔云,卞如潦,陈修主编.药理实验方法学[M].北京：人民卫生出版社.2002,817
[1]Kita I,Seki Y.Nakatani Y,etal.Corticotropin-releasing factor neurons in the hypothalamic paraventricular nucleus are involved in arousal yawning response of rats [J].Behavior Brain Research,2006,169(1):48-56.
[2]李爱林.谷氨酸的兴奋性毒性与脑创伤[J].国外医学.神经病学神经外科学分册,2005,犯(3)：231-234
[3]王尧,杜子威.神经生物化学与分子生物学[M].北京：人民卫生出版社,1997,200
[4]工学铭.精神与精神病的生物化学[M]北京：人民卫生出版社,2002,170
[5]Watson CJ,Lydic R,Baghdoyan HA.Sleep and GABA levels in the oral part of rat pontine reticular formation are decreased by local and systemic administration of morphine [J] Neuroscience,2007,144(1):375-386.
[6]Mohler H. GABAA receptors in central nervous system disease:anxiety,epilepsy,and insomnia [J] Journal of Receptor and Signal Transduction Research,2006,26(5-6):731-740.
[7]王尧,杜子威.神经生物化学与分子生物学[M].北京：人民卫生出版社,1997,186
[8]Talalaenko A N, Krivobok G K,StakhovsliY Iu Y.The role of monoam in-and acidergic mechanism of the hippocampus in anxiety stataes of different origins and their partieipation in the antiaversive effects of anxiolyties[J].FiziolZh Im IM Sechenova,1993,79(1):99-104
[9]Kaluff a V,Nutt D J.Role of GABA in anxiety and depression[J].Depress Anxiety,2007,24(7):495-517
[10]Duzzioni M, Calixto A V, Duarte F S,et al.Modulation of anxiety in rats evaluated in the elevated T-maze:evidence of the relation ship between substance P and diazepam[J].BehavBrain Res,2008,187(1):140-145
[11]周奇志,赵纪岚,蔡定均,等.电针对慢性情绪应激焦虑大鼠中枢单胺递质与γ-氨基丁酸失平衡的调节作用[J].中华中医药杂志,2008,23(10)：926-929
[12]王玉超,韩国玲,李海霞.丁螺环酮治疗焦虑症前后脑内神经递质的变化及意义[J].青海医药杂志,2008,38(9)：11-13
[13]RobertM, Sapolsky.Why stress is bad for your brain[J].Science,1996, 272(5276):749-750
[14]王尧,杜子威.神经生物化学与分子生物学[M].北京：人民卫生出版社,1997,419
[1]Ehlers MD, Zhang S, Bernhadt JP, et al. Inactivation of NMDA receptors by direct of Calmodul in with the NR1 subunit. Cell,1996,84(5):745-755./
[2]沈丽,张琳,南燕NMDA受体与中枢神经系统发育[J].生理科学进展,1999,30(1)：60-62.
[3]Watson CJ, Lydic R, Baghdoyan HA. Sleep and GABA levels in the oral part of rat pontine reticular formation are decreased by local and systemic administration of morphine [J] Neuroscience,2007,144(1):375-386.
[4]Mohler H. GABAA receptors in central nervous system disease:anxiety, epilepsy, and insomnia [J].Journal of Receptor and Signal Transduction Research,2006,26(5-6):731-740.
[1]Ehlers MD, Zhang S, Bernhadt JP, et al. Inactivation of NMDA receptors by direct of Calmodul in with the NR1 subunit. Cell,1996,84(5):745-755./
[2]沈丽,张琳,南燕NMDA受体与中枢神经系统发育[J].生理科学进展,1999,30(1)：60-62.
[3]Watson CJ, Lydic R, Baghdoyan HA. Sleep and GABA levels in the oral part of rat pontine reticular formation are decreased by local and systemic administration of morphine [J] Neuroscience,2007,144(1):375-386.
[4]Mohler H. GABAA receptors in central nervous system disease:anxiety, epilepsy, and insomnia [J].Journal of Receptor and Signal Transduction Research,2006,26(5-6):731-740.
[2]萨默斯·朗文当代英语大辞典[M].北京：商务印书馆,2004：60.
[3]张奇文.英汉双解精选医学词典[M].北京：人民卫生出版社,1990：465.
[4]American Psychiatric Association.Diagnostic and Ststistical Manual of Mental disorders.4th ed.Washington DC:APA 1994
[5]Murray CJ, Lopez AD.Altemative Projeetions of mortality and disability by cause 1990～2020.Global Burden of Disease Study [J] J.Lancet,1997,349: 1498-1504
[6]Ustun TB, Sartourius N. Mental illness in general healthcare. New York: Wiley,1995.323-375
[7]Kessler RC, Berglund P, Demler O.Lifetime prevalence and age-ofonset distributions of DSM-IV disorders in the National Comorbidity Survey Replication. Arch Gen Psychiatry 2005,62:593-602
[8]Pigott TA.Gender differences in the epidemiology and treatment of anxiety disorders.J Clin Psychiatry,1999,60(Supp118):4
[9]袁勇贵.焦虑障碍共病研究.[J].中国临床康复,2004,8(33)：7544-7547
[10]陈复平.神经症的流行病学调查[J].中华神经精神科杂志,1986,19(3)：301
[11]区丽明, 叶瑞繁, 陈剑等.广州地区综合医院门诊抑郁/焦虑障碍流行病学调查[J].中国临床心理学杂志,2009,17(1)：61
[12]向孟泽,单友荷,段诚凤,等12地区神经症流行病学调查[J].中华神经精神科杂志,1986,19(2)：87
[13]王欣,王守勇.酸枣仁汤临床与实验研究评述[J].中成药,2005,25(5)：414
[14]工泽帅,马建军,李学,等.神经内科门诊患者焦虑和抑郁症状的调查研究[J].中国实用神经疾病杂志,2009,12(1)：51-53
[15]Alonso J, Angermeyer MC, Bernert S, et al. Prevalence of mental disorders in Europe:results from the European Study of the Epidemiology of Mental Disorders (ESEMeD) project. Acta Psychiatr Scand 2004,109(suppl 420): 21-27
[16]Wittchen H-U, Nelson CB, Lachner G. Prevalence of mental disorders and psychosocial impairments in adolescents and young adults.Psychol Med 1998,28:109-126
[17]Murray CJ, Lopez AD.Alternative Projections of mortality and disability by cause 1990-2020.Global Burden of Disease Study [JJ.J.Lancet,1997,349: 1498-1504
[18]Leckman JF,Merikangas KR,Pauls DL,et al.Anxiety disorders and depression:contradictions between family study data and DSM-Ⅲ conventions.Am J Psych Iatry,1983,140(7):880
[19]Kendler KSJ。Neale MC,Kessler RC,eral.Majorde Pression and generalized anxiety disorder:Same genes,(Party)different environment[J].Arch Gen Psychiatry,1992,49:716-722
[20]Roy MA.Neale MC,Pedersen NL,etal.Atwinstudy of generalized anxiety disorder and major depression[J].Psyehol Med,1995,25:1037-1049
[21]Kendle KS, Walters EE.Neale MC.et al.The structure of the genetic and environment risk factors six major psychiatric disorders in women:Phobia, GAD.Panic disorder, bulimia major depression and aleoholism[J]. Areh Gen Psyehiatry,1995,52:374-383
[22]马震祥,刘玉局.广一泛性焦虑遗传效应和遗传方式研究[J].临床精神医学杂志,2006,16(6)～337
[23]Wu JC, Buehsbaum MS,Hershey T,et al.Positron emission tomography study of generalized anxiety disorder[J].Biol Psychiatry,1991,29:1181-1199
[24]万黎,刘建,吴文源,等.焦虑症患者平静及紧张状态局部脑血流灌注改变[J].中华核医学杂志,2002,22：206-208
[25]孙延强,赵相英,郭秀珍,等.焦虑症、抑郁症患者与正常健康人颅脑CT对比研究[J].中国行为医学科学,2005,12(14)：1082-1083
[26]王维治主编.神经病学.第5版.人民卫生出版社,2006：184-185
[27]Gerra G,Zaimovic A,Zambelli U,et al.Neuroendcrine responses to psychological stress in adolescents with anxiety disorder. Neuropsychobiology,2000,42:82
[28]Guimaraes FS, Carobrez AP, De Agniar JC, et al.Anxioly effect in the elevated plus-maze of the NMDA receptor antagonist AP7 mieroinjeeted into the dorsal periaqueduetalgrey[J].Psychopharmaeology(Beri),1991,103(1):91-94
[29]Matheus MG, Guimaraes FS.Antagonism of non-NMDA receptors in the dorsal periaqueduetal grey induces anxiolytic effeet in the elevated plus maze[J].Psychophamaeology(Berl),1997,132(1):14-18
[30]李东亮,张朝.基础神经生物学[M].北京：人民军医出版社,2006,122
[31]Sibile E,Pavlides C,Benke D, Toth M.Genetic inactivation of the Serotonin(lA) receptor alpha subuzlits,reduetion of GABA(A) receptor binding,and belzodiazePine-resistant anxiety[J].JNeurosci,2000,20:2758-2765
[32]Nutt DJ.Neurobiologieal mechanism in generealized anxiety disorder [J].J Clin Psychiatry,2001,62(Suppl):22-27
[33]Ballenger JC.Overview of different pharmaeotherapies for attaining remission in generalized anxiety disorder[J].J Clin Psyhiatry,2001,62(Supp119):11-19
[34]张瑞华,李丽琴,王惠芳,等.A型氨基丁酸受体结构及其有关药物[J].生命的化学,2003,23(6)：441-444
[35]Serra M, Pisu MG,Littera M, et al.isolation induced decreased in both the abundanee of neuroaetive steroids and GABAA receptor function in rat brain[J].J Neuroehem,2000,75:732-740
[36]Kathryn M, Connor, Jonathan R,et al.Genralized anxiety disorder:neurobiologieal and pharmaeotherapeutic persectives[J].Biological Psychiatry,1998,44(12):1286-1294
[37]Hoyer D,Clarke DE,Fozard JR,et al.1.International union of pharmaeoloy classification of receptors for 5-hydroxytryptamine(serotonin)[J].Pharmacol Rev,1994,46:157-203
[38]吴东辉,刘少文.广泛性焦虑障碍的神经生物学机制[J].国外医学·精神病学分册,2003,30(2)：124-126
[39]工振,肖泽萍,陈迁,等.焦虑症的生化病理机制探讨[J].临床精神医学杂志,2003,13(1)：1～3
[40]Baldwin D, Rudge S.The role of serotonin in depression and anxiety[J].Int ClinPsychoPharmacol,1995,9(suppl4):41-45
[41]谢健,高力舒.焦虑症艾森克个性问卷与血浆去甲肾上腺素、总皮质醇及白介素-2测定及相关分析[J].江西医药,2006,41(5)：266-268
[42]Kathryn M, Connor, Jonathan R.T.et al.Generalized anxiety disorden:neurobiological and Pharmaeotherapeutic perspectives[J].Biol Psychiatry,1998,44(12):1286-1294
[43]中华医学会精神科分会CCMD-3中国精神障碍分类与诊断标准[M].第3版.济南：山东科学技术出版社,2001,106～107
[44]王焕林.临床精神医学[M].北京：人民军医出版社,2003,324
[45]张德昌.医学药理学[M].北京：北京医科大学、中国协和医科大学联合出版社,1998.431.
[46]梁正英等.丁螺环酮在社交焦虑障碍治疗中的应用.贵阳医学院学报,2005(5)：445-446.
[47]翟金国等,曲唑酮与丁螺环酮治疗广泛性焦虑障碍的对照研究.中国新药与临床杂志,2008(11)：832-835
[48]邹万芹.新型抗抑郁药的临床应用及研究进展[J].中国药房,2008,19(14)： 1105.
[49]白秋江,郑敏.选择性5-羟色胺再摄取抑制剂致不良反应文献分析[J].中国药房,2007,18(20)：1583.
[50]孙松涛.柴胡加龙骨牡蛎汤加减治疗广泛性焦虑症72例[J].中医杂志,2000,41(2)：95.
[51]雷翔,柴胡加龙骨牡蛎汤临证举隅[J].中国民族民间医药,2010(9)：138.
[52]倪良玉,柴胡加龙骨牡蛎汤的临床应用体会[J].山西中医,2005(2)：58-59
[53]王守勇,谢鸣、王欣,酸枣仁汤组分与抗焦虑效用的相关性研究[J].中国实验方剂学杂志,2010(3).
[54]王业民、周建军、孔令东,半夏厚朴汤及其组成中药抗焦虑和抑郁研究进展[J].世界科学技术,2003(4).
[55]马行,库宝善,徐英,等.复方郁金胶囊抗抑郁抗焦虑的作用[J].中国临床康复,2006,10(15)：56.
[56]徐志伟,王文竹,苏俊芳,等.逍遥散和丹栀逍遥散抗焦虑作用的实验研究[J].广州中医药大学学报,2006,23(4)：330.
[57]贺玉琢.以群居干扰实验探讨加味逍遥散的抗焦虑作用及其活性成分(I)[J].国外医学-中医中药分册,2004,26(4)：250.
[58]加减逍遥散与氯硝西泮治疗焦虑症状群的对照观察[J].海峡药学,2004,(05).
[59]王玲姝,郑祖艳.针药结合对焦虑性神经症患者5-羟色胺及去甲肾上腺素 的影响[J].针灸临床杂志,2009,25(7)：13-14.
[60]罗文政,刘海静,梅尚英,等.靳三针治疗广泛性焦虑症的临床观察[J].中国中西医结合杂志,2007,27(3)：201-203.
[61]王贯民,针刺治疗广泛性焦虑症21例[J].河南中医,2007,27(7),55-56
[62]张涛,邵沛,陈妙.针药结合治疗焦虑症35例临床观察[J].针灸临床杂志,2006,22(12)：13-14.
[63]赵天辉,经络氧疗法治疗焦虑障碍60例疗效观察[J].新中医2006,38(2)：66-67.
[64]霍小宁,耳压法治疗广泛性焦虑28例临床观察[J].甘肃中医,2005,18(4)：30.
[65]太鑫,针灸结合心理疗法治疗广泛性焦虑障碍的临床观察[J].中医药信息,2010,27(2)：72-73.
[66]沈莉,颜红,冯辉,针药结合治疗广泛性焦虑症临床观察[J].上海针灸杂志,2007,26(3)：3-4
[67]黄奕,赵长鹰.浅述失眠症的心理治疗方法[J].陕西中医,2005,26(5)：479
[68]胡师焕.《内经》中有关心理学的理论观点[J].湖北中医学院学报,2004,6(3)：24
[1]中华医学会精神科分会.中国精神障碍分类与诊断标准[M].3版.济南：山东科学技术出版社,2001：106-107.
[2]张明园.精神科评定量表手册[M].长沙：湖南科学技术出版社,2003.
[3]中药新药临床研究指导原则(试行)[S].北京,中国医药科技出版社.2002.
[4]Kessler RC, Berglund P, Demler O.Lifetime prevalence and age-ofonset distributions of DSM-Ⅳ disorders in the National Comorbidity Survey Replication. Arch Gen Psychiatry 2005,62:593-602.
[5]熊曼琪.伤寒论[M].北京：中国中医药出版社,2007：202.
[6]吴斌龙.桂枝汤方证发微[J].辽宁中医药大学学报,2011,13(2)：99-100.
[7]贺一新,赵素霞,崔瑛.酸枣仁抗焦虑活性物质分析[J].中药 材,2010,33(2)：229-231.
[8]傅晶,张东明,陈若芸.远志属植物的皂苷类成分及其药理作用研究进展[J].中草药,2006,37(1)：144-146.
[9]Li X,Matsumoto K,Murakami Y,et al.europrotective effects of Polygonum multiflorum on nigroatriatal dopaminergic degeneration induced by paraquat and maneb in mice[J].Pharmacol Biochem Behav,2005,82(2):345-352.
[10]王维勋,孙付军,李芳,张希林,张军.柴胡加龙骨牡蛎汤对焦虑模型大鼠单胺类递质的影响[J].中药新药与临床药理,2008,19(5)：340-342.
[1]Pellow S, Chopin P, File SE, et al. Validation of open-closed arm entries in an elevated p lus-maze as a measure of anxiety in the rat[J]. Journal of Neuroscience Methods,1985,14 (3):149-167
[2]Montgomery S.Persistent c-fos expression in the brains of mice with soeial stress[J].Neurosei-Res,1996,26(2):157
[3]Holmes A,Rodgers R.J.Responses of Swiss-Webster mice to repeated plus-maze experience:further evidence for a qualitative shift in emotional state.Pharmacology Biochemistry and Behavior,1998,60(2):473-488
[4]Gard PR,Haigh SJ,Cambursano PT,et al.Strain differences in anxiolytic effects of losartan in the mouse.Pharmacology Biochemistry and Behavior,2001,69(1-2): 35-40
[5]Pellow S,Chopin P,File SE,et al.Validation of open:closed arm entries in an elevated plus-maze as a measure of anxiety in the rat[J].J Neurosci Methods,1985,14(3):149-167
[6]Lister RG.The use of a plus-maze to measure anxiety int the miee [J].Psycho Pharmaeology (Berl),1987,92(2):180-185
[7]Lister RG.The use of a plus-maze to measure anxiety in the mice [J].Psvchophaimacolony(Berl),1987,92(2):180-185
[8]程九清,李勇辉,隋南.基于啮齿动物的决策行为研究及其脑机制[J].心理科学杂志,2008,16(5)：721—725
[9]徐叔云,卞如潦,陈修主编.药理实验方法学[M].北京：人民卫生出版社.2002,817
[1]Kita I,Seki Y.Nakatani Y,etal.Corticotropin-releasing factor neurons in the hypothalamic paraventricular nucleus are involved in arousal yawning response of rats [J].Behavior Brain Research,2006,169(1):48-56.
[2]李爱林.谷氨酸的兴奋性毒性与脑创伤[J].国外医学.神经病学神经外科学分册,2005,犯(3)：231-234
[3]王尧,杜子威.神经生物化学与分子生物学[M].北京：人民卫生出版社,1997,200
[4]工学铭.精神与精神病的生物化学[M]北京：人民卫生出版社,2002,170
[5]Watson CJ,Lydic R,Baghdoyan HA.Sleep and GABA levels in the oral part of rat pontine reticular formation are decreased by local and systemic administration of morphine [J] Neuroscience,2007,144(1):375-386.
[6]Mohler H. GABAA receptors in central nervous system disease:anxiety,epilepsy,and insomnia [J] Journal of Receptor and Signal Transduction Research,2006,26(5-6):731-740.
[7]王尧,杜子威.神经生物化学与分子生物学[M].北京：人民卫生出版社,1997,186
[8]Talalaenko A N, Krivobok G K,StakhovsliY Iu Y.The role of monoam in-and acidergic mechanism of the hippocampus in anxiety stataes of different origins and their partieipation in the antiaversive effects of anxiolyties[J].FiziolZh Im IM Sechenova,1993,79(1):99-104
[9]Kaluff a V,Nutt D J.Role of GABA in anxiety and depression[J].Depress Anxiety,2007,24(7):495-517
[10]Duzzioni M, Calixto A V, Duarte F S,et al.Modulation of anxiety in rats evaluated in the elevated T-maze:evidence of the relation ship between substance P and diazepam[J].BehavBrain Res,2008,187(1):140-145
[11]周奇志,赵纪岚,蔡定均,等.电针对慢性情绪应激焦虑大鼠中枢单胺递质与γ-氨基丁酸失平衡的调节作用[J].中华中医药杂志,2008,23(10)：926-929
[12]王玉超,韩国玲,李海霞.丁螺环酮治疗焦虑症前后脑内神经递质的变化及意义[J].青海医药杂志,2008,38(9)：11-13
[13]RobertM, Sapolsky.Why stress is bad for your brain[J].Science,1996, 272(5276):749-750
[14]王尧,杜子威.神经生物化学与分子生物学[M].北京：人民卫生出版社,1997,419
[1]Ehlers MD, Zhang S, Bernhadt JP, et al. Inactivation of NMDA receptors by direct of Calmodul in with the NR1 subunit. Cell,1996,84(5):745-755./
[2]沈丽,张琳,南燕NMDA受体与中枢神经系统发育[J].生理科学进展,1999,30(1)：60-62.
[3]Watson CJ, Lydic R, Baghdoyan HA. Sleep and GABA levels in the oral part of rat pontine reticular formation are decreased by local and systemic administration of morphine [J] Neuroscience,2007,144(1):375-386.
[4]Mohler H. GABAA receptors in central nervous system disease:anxiety, epilepsy, and insomnia [J].Journal of Receptor and Signal Transduction Research,2006,26(5-6):731-740.
[1]Ehlers MD, Zhang S, Bernhadt JP, et al. Inactivation of NMDA receptors by direct of Calmodul in with the NR1 subunit. Cell,1996,84(5):745-755./
[2]沈丽,张琳,南燕NMDA受体与中枢神经系统发育[J].生理科学进展,1999,30(1)：60-62.
[3]Watson CJ, Lydic R, Baghdoyan HA. Sleep and GABA levels in the oral part of rat pontine reticular formation are decreased by local and systemic administration of morphine [J] Neuroscience,2007,144(1):375-386.
[4]Mohler H. GABAA receptors in central nervous system disease:anxiety, epilepsy, and insomnia [J].Journal of Receptor and Signal Transduction Research,2006,26(5-6):731-740.