安神方治疗失眠症的疗效观察及相关机制研究
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摘要
背景
自古以来,人类就对睡眠和梦产生了浓厚的兴趣。从西方的佛洛依德所著《梦的解析》开始,人类对睡眠和梦的探究从来没有停止过。睡眠作为人生命的重要组成部分,对人体保持充沛的精力体力,维持神经系统、内分泌系统、免疫系统功能正常均具有重要意义。
近年来,经济社会变革加速,人们的生活工作节奏均大幅加快,负面情绪引起的精神紧张焦虑、抑郁更是频繁发生,导致失眠症发生日益频繁。失眠(insomnia)是指睡眠的始发(sleep onset)和睡眠维持(sleep maintenance)发生障碍,致使睡眠质量不能满足个体生理需要而明显影响患者白天活动(如白天困倦乏力、记忆力注意力下降等)的一种睡眠障碍综合征。
现代医学对失眠的研究发现睡眠的发生和维持与视交叉上核、下丘脑、脑干网状上行系统以及松果体等部位,与乙酰胆碱(ACH)、去甲肾上腺素(NE)、多巴胺(DA)、单胺类、氨基酸类神经递质以及退黑激素(melatonin,MT)、前列腺素(Prostaglandin,PG)、白介素(interleukin,IL)等物质,均存在密切联系。
目前失眠症的临床治疗仍以BZD类镇静催眠药物为主,在改善失眠症状的同时,往往伴有不同程度毒副作用并容易形成药物依赖。祖国中医药的整体辩证论治方法,调整人体机能状态的同时达到改善睡眠的治疗方法,为失眠症患者带来了福音。导师陈宝田教授在研读中医经典《伤寒论》过程中,结合临床实践在“柴胡桂枝汤”的基础上创立了“安神方”,治疗失眠症疗效确切,同时对改善患者焦虑、抑郁等精神疾患办有疗效。在前期临床观察的基础上,我们设计本实验旨在系统地观察安神方治疗失眠症的临床疗效,并进一步通过动物实验探讨安神方治疗失眠症的作用机理,为安神方的开发提供可靠的理论和临床依据。
第一章安神方治疗失眠症临床观察
目的通过临床观察探讨安神方治疗失眠症的临床效果。观察失眠症症侯疗效,临床整体疗效以及多导睡眠图(PSG)检查客观结果,客观的评价安神方治疗失眠症的效果。
方法
1、病例与分组符合纳入标准的失眠症患者80例,随机方法分为安神方治疗组和枣仁安神胶囊对照组;
2、用药治疗组安神方颗粒剂每日1剂,1次/晚;对照组枣仁安神胶囊每次4粒,1次/晚。睡前1小时服药,时间4周。(安神方,由南方医院中药房提供,由广东一方制药生产,许可证号:粤zbx20060360;枣仁安神胶囊,由南方医院药房提供,贵州同济堂制药有限公司生产,国药准字号Z20010033);
3、中医症候疗效评价在中医症候评分基础上,比较治疗前后主要症状(睡眠时间,入睡困难,易醒,多梦,困倦乏力)改善等级,采用非参数检验方法,分析评价各组症状的改善;
4、临床整体疗效评价根据治疗前后总评分,计算反应总体改善情况的“疗效指数”,评定每例患者改善等级,以此评价两组治疗的整体疗效;
5、临床多导睡眠图疗效评价以分钟(min)为单位记录两组治疗前后总睡眠时间(TST)、睡眠潜伏时间、觉醒次数,浅睡眠时间(S1+S2),深睡眠时间(S3+S4)及REM睡眠时间。分别进行治疗前差异比较(两独立样本t检验)、组内治疗前后症状比较(两配对样本t检验)、治疗后组间症候疗效比较(协方差分析);
6、安全性评价治疗前后检查体温,心率,血压,呼吸;血、尿、便常规检查;肝、肾功能检查;心电图检查;
7、统计分析采用统计软件SPSS13.0进行统计学处理和分析。实验数据计量资料采用均数加减标准差((?)±S)表示。治疗前后剂量资料比较采用两配对样本t检验;两组间计量资料比较采用两独立样本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作为差异有显著性差异的界限。
结果
1、一般资料两组在性别(χ~2=0.213,P=0.646)、年龄(F=1.798,P=0.184)、病程(t=1.031,P=0.306),方面比较无显著性差异(P均＞0.05),具有可比性;
2、症候疗效评价两组对“睡眠时间”的疗效对照(Z=-0.633,P=0.527),观察两组对“入睡困难”的疗效对照(Z=-1.716,P=0.086),观察两组对“多梦”的疗效对照(Z=-1.115,P=0.265),均无显著性差异;观察两组对“易醒”症状的疗效对照(Z=-2.285,P=0.022),对“困倦乏力”症状的疗效对照(Z=-2.856,P=0.004),均有显著性差异;
3、整体疗效评价两组治疗对失眠症整体疗效的比较(Z=-2.130,P=0.033),有显著性差异,治疗组总体疗效优于对照组;
4、多导睡眠图(PSG)疗效评价
治疗前两组间,对“总睡眠时间”疗效比较(t=-0.153,P=0.879);“睡眠潜伏时间”疗效比较(t=-0.400,P=0.691);“觉醒次数”疗效比较(t=-0.733,P=0.467);“浅睡眠时间”疗效比较(t=0.290,P=0.773);“深睡眠时间”疗效比较(t=.1.021,P=0.312);“REM睡眠时间”疗效比较(t=.0.425,P=0.673),均无显著性差异。说明两组具有可比性;
治疗前后,治疗组内各症状比较结果:对“总睡眠时间”疗效比较(t=-3.534,P=0.002),“觉醒次数”疗效比较(t=2.753,P=0.011),“深睡眠时间”疗效比较(t=-4.853,P=0.000),“REM睡眠时间”疗效比较(t=-4.596,P=0.000),均有显著性差异;对“睡眠潜伏时间”疗效比较(t=1.572,P=0.129),“浅睡眠时间”疗效比较(t=0.409,P=0.686),无显著性差异。说明治疗组具有“增加总睡眠时间、深睡眠时间、REM睡眠时间,减少觉醒次数”的效果;
对照组内各症状比较:对“总睡眠时间”疗效比较(t=-2.757,P=0.011),“睡眠潜伏时间”疗效比较(t=2.324,P=0.029),“浅睡眠时间”疗效比较(t=-2.396,P=0.025),均有显著性差异;“觉醒次数”疗效比较(t=1.821,P=0.081),“深睡眠时间”疗效比较(t=-1.846,P=0.077),“REM睡眠时间”疗效比较(t=-1.028,P=0.314),均无显著性差异。说明对照组具有“增加总睡眠时间、缩短睡眠潜伏期、增加浅睡眠时间”的效果;
治疗后两组间疗效比较,对“总睡眠时间”疗效比较(F=0.394,P=0.533),对“睡眠潜伏时间”疗效比较(F=1.633,P=0.208),两组对“觉醒次数”的疗效比较(F=1.053,P=0.310),均无显著性差异;对“浅睡眠时间”的疗效(F=9.042,P=0.004),对“深睡眠时间”的疗效比较(F=4.887,P=0.032),对“REM睡眠时间”的疗效比较(F=8.441,P=0.006),均有显著性差异;说明治疗组增加深睡眠时间、REM睡眠时间的疗效优于对照组;对照组增加浅睡眠时间疗效优于治疗组。
结论
1、安神方治疗对改善“易醒,困倦乏力”症状的疗效优于对照组,治疗失眠症的整体疗效优于对照组;
2、安神方对“总睡眠时间、觉醒次数、深睡眠时间、REM睡眠时间”症状的改善,具有显著疗效;与对照组比较,改善“深睡眠时间、REM睡眠时间”的疗效显著。其对睡眠结构的调整主要是增加了“深睡眠时间”及“REM睡眠时间”;致使“总睡眠时间”增加;
3、安全性指标观察各项检查指标无异常。
第二章安神方治疗失眠症的相关机制研究
一、安神方对失眠造模大鼠中枢氨基酸类神经递质的影响
目的观察安神方对大鼠中枢氨基酸类神经递质含量的影响,探讨其治疗失眠症的机理。
方法
1、实验动物与分组清洁级Wistar大鼠60只,体重(200±20g),南方医院实验动物中心提供,许可证号:(粤)2006-0015。按随机方法分成高剂量组、中、低剂量组和生理盐水对照组,共4组;
2、给药高剂量组大鼠每天按40g.Kg~(-1)灌胃,中剂量组大鼠每天按20g.Kg~(-1)灌胃,低剂量组大鼠每天按10g.Kg~(-1)灌胃,生理盐水组给予等量生理盐水灌胃。失眠造模同时连续给药15天;
3、失眠大鼠模型制备方法本实验采用‘限制运动法'制作大鼠失眠模型:将大鼠置于自制的大鼠运动限制仪内,通过移动插片控制大鼠的活动空间。每日束缚制动1次,第一次4h,其后每次增加1h,连续15d完成。大鼠在制动期间禁食、禁水;
4、中枢氨基酸类神经递质检测方法处死并分离大鼠海马及下丘脑。称重,玻璃匀浆器匀浆化;离心取上清液,采用“高效液相色谱荧光检测法”检测中枢氨基酸类神经递质;
5、仪器和试剂
仪器及试剂:DIONEX Summit高效液相色谱仪(美国DIONEX公司),Chromeleon色谱工作站(美国DIONEX公司),RF2000荧光检测器(美国DIONEX公司),PHS-2C精密酸度计(上海理达仪器厂),冷冻离心机(Avanti 30),电子天平(BA 110S)。谷氨酸(Glu)、γ-氨基丁酸(GABA)、OPA(内含2-ME)均购自美国SIGMA公司,批号分别为43907190、026K07361、116K5005;乙腈购自美国TEDIA公司;其余试剂均为市售分析纯;
6、实验数据计量资料采用((?)±S)表示;采用统计软件SPSS13.0进行统计学描述和分析。多组间比较采用单因素方差分析(One-Way ANOVA)检验;方差齐时采用LSD方法检验;方差不齐时采用非参数Kruskal-Wallis检验。所有统计数据均以P＜0.05作为差异有显著性的界限。
结果
1、海马γ-氨基丁酸(GABA)含量高剂量组与生理盐水组比较(P=0.002),中剂量组与生理盐水组比较(P=0.017),低剂量组与生理盐水组比较(P=0.045),均有显著性差异,治疗组GABA含量高于对照组;
2、海马谷氨酸(GLU)含量:高剂量组与生理盐水组比较(P=0.011),中剂量组与生理盐水组比较(P=0.045),均有显著性差异,治疗组GLU含量低于对照组;低剂量组与生理盐水组比较(P=0.566),无显著性差异;
3、下丘脑γ-氨基丁酸(GABA)含量:高剂量组与生理盐水组比较(P=0.001),中剂量组与生理盐水组比较(P=0.028),均有显著性差异,治疗组GABA含量高于对照组;低剂量组与生理盐水组比较(P=0.166),无显著性差异;
4、下丘脑谷氨酸(GLU)含量:高剂量组与生理盐水组比较(P=0.015),有显著性差异,治疗组GLU含量低于对照组;中剂量组与生理盐水组比较(P=0.103),无显著性差异;低剂量组与生理盐水组比较(P=0.798),无显著性差异。
结论安神方治疗组能增加大鼠海马、下丘脑中γ-氨基丁酸(GABA)含量,降低谷氨酸(GLU)含量;调节中枢氨基酸类神经递质是安神方治疗失眠症的机制之一。
二、安神方对睡眠剥夺大鼠血清皮质醇的影响
目的观察安神方对应激条件下动物的焦虑症状的调节作用。
方法
1、动物及分组清洁级Wistar大鼠30只,8周龄,雄性,体重(300±30g),南方医院实验动物中心提供,许可证号:(粤)2006-0015。随机方法分为2组,每组15只;
2、试验药物中药安神方,由南方医院中药房提供。治疗组大鼠每天按20g.Kg~(-1)灌胃(剂量按成人换算),生理盐水组给予等量生理盐水;每天灌胃1次;
3、睡眠剥夺大鼠模型制备采用“改良多平台睡眠剥夺法”进行睡眠剥夺(sleep deprivation,SD)。室温保持20-25℃,正常昼夜光照,每日更换睡眠剥夺用水,保持自由进食条件;
4、试验步骤:持续睡眠剥夺72h,两组分别在睡眠剥夺(SD)Oh,24h,48h和72h时,记录各组动物体重,并采血检测血清皮质醇。血清皮质醇测定方法:于早晨7时经尾静脉取血0.5ml,置离心管中以3000r/min离心10min,取上清液保存于低温冰箱(-60℃)待检测;
5、资料整理与统计学方法:采用统计软件SPSS13.0进行统计学处理和分析。治疗前后指标差值采用协方差分析(analysis of covariance,ANCOVA),不同时间点数据变化采用重复测量数据的方差分析(ANOVA for the repeated measures)和独立样本t检验进行比较(Independent-Samples T Test)。所有统计数据分析均以P＜0.05作为差异有显著性的界限。
结果
1、两组治疗前后体重变化比较(F=79.682,P=0.000),有显著性差异;
2、第1,2天治疗组和对照组的体重比较无显著性差异(第1天:t=-0.429,P=0.671;第2天:t=1.835,P=7.077),而在第3,4天两个时间点上,两组的体重比较均有显著性差异(第3天:t=3.675,P=0.001;第4天:t=5.675,P=0.00)。治疗组和对照组体重在不同时间点之间差异有显著性差异(F=460.01,P=0.000)。分组与时间之间存在交互作用(F=70.22,P=0.000);
3、两组治疗前后皮质醇含量比较(F=70.601,P=0.000),有显著性差异;
4、第1,2天治疗组和对照组的血清皮质醇比较无显著性差异(第1天:t=-0.372,P=0.712;第2天:t=-0.699,P=0.490),而在第3,4天两个时间点上,两组的血清皮质醇比较均有显著性差异(第3天:t=-3.247,P=0.003;第4天:t=-4.770,P=0.000)。治疗组和对照组血清皮质醇在不同时间点之间差异有显著性差异(F=128.94,P=0.000)。分组与时间之间存在交互作用(F=29.846,P=0.000)。
结论
1、治疗组体重减轻速度慢于生理盐水对照组;
2、治疗组血清皮质醇含量增加的速度慢于生理盐水对照组;
3、安神方治疗可以减轻睡眠剥夺大鼠的焦虑症状。
Background
since the ancient times,the humanity on has had the strong interest to the sleep and the dream.The humanity has not stopped regarding the sleep and the dream inquisition since Floyd's "Dream Analysis.As an important constituent,sleep play a important role not only in maintaining human's physiology healthy,but also psychological is.Normal sleep maintains the abundant energy physical strength to the human body,significance of sleep in relation to maintenance nervous system,the endocrine system,and the immune system function normal.
In recent years,anxious and depression from accelerated society transformand and people's life rhythm,made the insomnia occur frequently.Insomnia is a kind of sleep barrier synthesis that something wrong in the onset and maintenance of sleep, far from satisfying the individual physiological needs,affecting the patient's life(for example the daytime sleepy asthenia,remembrance breakdown and so on).The modern medicine discovered the occurrence and the maintenance of sleep had a correlated with not only the suprachiasmatic nucleus,hypothalamus,ascending reticular activating system and apophysis cerebri,but also acetylcholin(ACH), noradrenalin(NE),dihydroxyphenyl ethylamine(DA),the single-amines neurotransmitt e,the amino acid neurotransmitte as well as melatonin,prostaglandin,interleukin.
At present,the main drug to the insomnia is still BZD sedative hypnotics,which bring adverse reaction and drug dependence as well as treat insomnia.The differentiation of Traditional Chinese medicine,adjusts the human performance condition,achieves the improvement of sleep,has brought the gospel for the insomnia patient.Famous physician Chen Baotian of traditional Chinese medicine created "AnShenfang" through reading the Chinese medicine classics "Shanghanlun",and confirmed curatively for a long time in clinical work.simultaneously improved the patient's anxious and despondent.on the base of clinical observation foundation,we designs this experiment to observate the the therapeutic effect of the AnShenfang systematically,and further discusses the mechanism of AnShenfang through the animal experimentation,provides the reliable theory and the clinical basis for the development of AnShenfang.
chapter 1.AnShenfang treats the insomnia clinical observation
Objective To objectively estimate the therapical effect of "AnShenfang" in treating insomnia of Liver-qi and Changes into Liver-fire through the clinical overall curative effect as well as the Polysomnogram(PSG).
Methods
1.Clinical curative effect observation:80 cases are selected and randomly devided into two groups:the AnShenfang group and the control group;
2.According to level of Chinese medicine symptom graduation foundation, compared improvement of main symptom(sleep time,difficulty falling asleep,easily awakening,dreaminess,fatigue),useing the non-parametric test;
3.According to grades around the treatment,calculate "the curative effect index", evaluates rank of improvement in each patient,appraises the total curative effect;
4.Evaluating the therapical effect though Polysonmogram(PSG),records each data(total sleep time,sleep latency time,awareness times,shallow sleep time,deep sleep time,REM sleep time).Objectively appraisal curative effect.
Rusults
1.Clinical observation:there was no significant differences in gender,age and course of disease in two groups(P>0.05),the two groups are comparable;
2.Symptom curative effect appraises:there was no significant differences in "sleep time(Z=-0.633,P=0.527),difficulty falling asleep(Z=-1.716,P=0.086), dreaminess(Z=-1.115,P=0.265)" between two groups;there was significant difference in "easily awakening(Z=-2.285,P=0.022),fatigue(Z=-2.856,P= 0.004) "between two groups;
3.The total curative effect appraises:there was significant difference in total curative effect(Z=-2.130,P=0.033),the treatment group total curative effect surpasses the control group;
4.The polosomnography(PSG) curative effect appraisal:
Before treatment,there was no significant difference in total sleep time compares(t=-0.153,P=0.879);sleep latency time compares(t=-0.400,P=0.691); awareness times compares(t=-0.733,P=0.467);shallow sleep time compares (t=0.290,P=0.773);the deep sleep time compares(t=-1.021,P=0.312);the REM sleep time compares(t=-0.425,P=0.673)between two groups.Two groups are comparable;
Around treatment,comparison in treatment group:there were significant difference in the total sleep time compares(t=-3.534,P=0.002),awareness times compare(t=2.753,P=0.011),the deep sleep time compares(t=-4.853,P=0.000), REM sleep time compares(t=-4.596,P=0.000),there was no significance difference Regarding the sleep latency time compares(t=1.572,P=0.129),and the shallow sleep time compares(t=0.409,P=0.686).Ttreatment group has effect on"increasing the total sleep time,the deep sleep time,the REM sleep time,reduces awareness times;
Symptoms comparison in control group:there were significant differences in the total sleep time compares(t=-2.757,P=0.011),the sleep latency time compares (t=2.324,P=0.029),the shallow sleep time compares(t=-2.396,P=0.025);there was no significant differences in awaken times compares(t=1.821,P=0.081),deep sleep time compares(t=-1.846,P=0.077),REM sleep time compares(t=-1.028,P=0.314), Control group has effect on"the increase sleep total time,the reduce sleep latency time,increases the shallow sleep time";
After the treatment,there were significant differences in the shallow sleep time compares(F=9.042,P=0.004),the deep sleep time compares(F=4.887,P=0.032),the REM sleep time compare(F=8.441,P=0.006) between two groups.there was no in the total sleep time compare(F=0.394,P=0.533),the sleep latency time compare (F=1.633,P=0.208),awareness times compare(F=1.053,P=0.310),increaseing the deep sleep time,the REM sleep time curative effect in treatment group surpasses the control group;The control group increasing the shallow sleep time curative effect surpass the treatment group.
Conclusion
1.Curative effect observation:the effect of "easily awakening,fatigue" in the treatment group surpasses the control group's,and the overall curative effect of treat group surpass the control group;
2.Polysomnogram(PSG) observation:The AnShenfang has significant effect on "the total sleep time,awareness times,the deep sleep time,the REM sleep time", there were significant differences in the "deep sleep time,the REM sleep time" Compared with the control group.The treat group mainly increased "total sleep time"through the increasing of"the deep sleep time" and "the REM sleep time";
3.The treatment security,each inspection target does not have exceptionally.
Chapter 2 laboratory explore of "AnShenfang" treat insomnia
1.The Effect of "AnShenFang"on Central Neurotransmitters of insomnic Rats
Objective Observe the effect of "AnShenFang"on Central Neurotransmitters of insomnic Rats,discusses the mechanism to treat the insomnia.
Methods
1.The experimental animal and group:60 Wistar rats,the body weight (200±20g),south the hospital experimental animal center provides,permit number: (Guangdong) 2006-0015.were randomly divided into the high dose group,middle dose group,lowdose group and the saline control group;
2.Administration:40g.Kg~(-1) for the high dose group.20g.Kg~(-1) for the middle dose group,and 10g.Kg~(-1) for the low dose group and the saline for the control group;meanwhile,made the insomnia model for 15 day;
3.insomnia rats model preparation:making insomnia rats model through limiting rats'movement:rats were Put in self-maderats movement limit meter,controls the rat's active space through the motion film threading,applies the brake 1 time each day, first time 4h,increases 1h each time after that,sustaining for 15d,diet and water were prohibited in this period;
4.examination of the central nervous:separate both hippocampus and hypothalamus of the rat.weighting,glass homogenizer refining;centrifugalization, quantitating through the high performance liquid chromatogram(HPLC);
5.Uses statistical software SPSS13.0 to 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.
Rusults
1.Gamma-amino-butyric acid(GABA) content in hippocampus:there were significant differences among the high dose group and saline control group compare (P=0.002),the middle dose group and saline control group compare(P=0.017),the low dose group and saline control group compare(P=0.045);
2.Glutanic acid(GLU) content in hippocampus:there were significant differences among the high dose group and the saline control group compare (P=0.011),the middle dose group and the saline control group compare(P=0.045); there was no significant difference between the low dose group and saline control group compare(P=0.566);
3.Gamma-amino-butyric acid(GABA) content in hypothalamus:there were significant differences among the high dose group and saline control group compare (P=0.001),the middle dose group and the saline control group compare (P=0.028);there was no significant difference between the low dose group and the saline control group compare(P=0.166);
4.Glutanic acid(GLU) content in hypothalamus:there was significant difference between the high dose group and the saline control group compare (P=0.015);there were no significant differences between the middle dose group and the saline control group compare(P=0.103),as well as the low dose group and the saline control group compare(P=0.798).
Conclusion The AnShenfang can elevate the Gamma-amino-butyric acid (GABA) content,reduces the glutanic acid(GLU) content both hippocampus and hypothalamus of the rats.
2.The Effect of "AnShenfang"on serum cortisol of Sleep Deprivated Rats
Objective To observe the effect of "AnShenfang" on Serum cortisol of Sleep Deprivated Rats.
Methods
1.The experimental animal and group:30 rats were randomized divided into 2 groups:treat group and normal control group,each group had 15 animals.Treat group were fed by "AnShenFang"and normal control group were fed by normal saline for 3 days,then sleep deprivated for 72 hours,test the body weight and content of serum cortisol on sleep deprivation 0h,24h,48h and 72h;
2.Administration "AnShenfang"20g.Kg~(-1) for the treat group,and the saline for the control group;
3.The rats sleep deprives model preparation:use"the multi-platform sleep deprivation methord" prepare sleep deprivation model(sleep deprivation,SD).The room temperature maintenancing 20-25℃,replacing the the water used every day, maintenancing free feed condition;
4.Testing methord:sleep depriving for 72h Continuesly,records each group of animal body weight,and detects blood serum cortisol on 0h,24h,48h and 72h.Blood serum cortisol determination method:on7 AM draws blood 0.5ml from the tail vein, centrifuge at 3000r/min for 10min,preserve serumin the low temperature refrigerator for detect(-60℃);
5.Uses statistical software SPSS13.0 to detail the data.use the analysis of covariance(ANCOVA)around test;uee the ANOVA for the repeated measures and Independent-Samples T Test to test the data on the different time All statistical analysis has the significance boundary as P<0.05.
Rusults
1.There were significant differences in the body weight of the two groups compare(F=79.682,P=0.000);
2.There was no significant difference on body weight at the 1,2nd day between the treat group and the control group's compares(1st day:t=-0.429,P=0.671;2nd days:t=1.835,P=7.077),but there were significant differences on bodu weight between weo group on the 3,4th day(3rd days:t=3.675,P=0.001;4th days:t=5.675, P=0.00).there was significant difference on body weight on different time(F=460.01, P=0.000).Between the grouping and the time has the correlation(F=70.22,P=0.000);
3.There were significant differences in the serum cortisol of the two groups compare(F=70.601,P=0.000);
4.There was no significant difference on content of serum cortisol at the 1,2nd day between the treat group and the control group's compares(1st day: t=-0.372,P=0.712;2nd days:t=-0.699,P=0.490),but there were significant differences on content of serum cortisol between two groups on the 3,4th day(3rd days:t=3.675, P=0.001;4th days:t=5.675,P=0.00).there was significant difference on content of serum cortisol on different time(F=460.01,P=0.000).Between the grouping and the time has the correlation(F=70.22,P=0.000).
Conclusion
1.The speed of body weight of Treat group was slow compared with saline control group;
2.The speed of content of serum cortisol of treat group was slower than compared with saline control group;
3.Anshenfang granules can elivate the rat's anxious symptom which reduce the sleep deprive.
自古以来,人类就对睡眠和梦产生了浓厚的兴趣。从西方的佛洛依德所著《梦的解析》开始,人类对睡眠和梦的探究从来没有停止过。睡眠作为人生命的重要组成部分,对人体保持充沛的精力体力,维持神经系统、内分泌系统、免疫系统功能正常均具有重要意义。
近年来,经济社会变革加速,人们的生活工作节奏均大幅加快,负面情绪引起的精神紧张焦虑、抑郁更是频繁发生,导致失眠症发生日益频繁。失眠(insomnia)是指睡眠的始发(sleep onset)和睡眠维持(sleep maintenance)发生障碍,致使睡眠质量不能满足个体生理需要而明显影响患者白天活动(如白天困倦乏力、记忆力注意力下降等)的一种睡眠障碍综合征。
现代医学对失眠的研究发现睡眠的发生和维持与视交叉上核、下丘脑、脑干网状上行系统以及松果体等部位,与乙酰胆碱(ACH)、去甲肾上腺素(NE)、多巴胺(DA)、单胺类、氨基酸类神经递质以及退黑激素(melatonin,MT)、前列腺素(Prostaglandin,PG)、白介素(interleukin,IL)等物质,均存在密切联系。
目前失眠症的临床治疗仍以BZD类镇静催眠药物为主,在改善失眠症状的同时,往往伴有不同程度毒副作用并容易形成药物依赖。祖国中医药的整体辩证论治方法,调整人体机能状态的同时达到改善睡眠的治疗方法,为失眠症患者带来了福音。导师陈宝田教授在研读中医经典《伤寒论》过程中,结合临床实践在“柴胡桂枝汤”的基础上创立了“安神方”,治疗失眠症疗效确切,同时对改善患者焦虑、抑郁等精神疾患办有疗效。在前期临床观察的基础上,我们设计本实验旨在系统地观察安神方治疗失眠症的临床疗效,并进一步通过动物实验探讨安神方治疗失眠症的作用机理,为安神方的开发提供可靠的理论和临床依据。
第一章安神方治疗失眠症临床观察
目的通过临床观察探讨安神方治疗失眠症的临床效果。观察失眠症症侯疗效,临床整体疗效以及多导睡眠图(PSG)检查客观结果,客观的评价安神方治疗失眠症的效果。
方法
1、病例与分组符合纳入标准的失眠症患者80例,随机方法分为安神方治疗组和枣仁安神胶囊对照组;
2、用药治疗组安神方颗粒剂每日1剂,1次/晚;对照组枣仁安神胶囊每次4粒,1次/晚。睡前1小时服药,时间4周。(安神方,由南方医院中药房提供,由广东一方制药生产,许可证号:粤zbx20060360;枣仁安神胶囊,由南方医院药房提供,贵州同济堂制药有限公司生产,国药准字号Z20010033);
3、中医症候疗效评价在中医症候评分基础上,比较治疗前后主要症状(睡眠时间,入睡困难,易醒,多梦,困倦乏力)改善等级,采用非参数检验方法,分析评价各组症状的改善;
4、临床整体疗效评价根据治疗前后总评分,计算反应总体改善情况的“疗效指数”,评定每例患者改善等级,以此评价两组治疗的整体疗效;
5、临床多导睡眠图疗效评价以分钟(min)为单位记录两组治疗前后总睡眠时间(TST)、睡眠潜伏时间、觉醒次数,浅睡眠时间(S1+S2),深睡眠时间(S3+S4)及REM睡眠时间。分别进行治疗前差异比较(两独立样本t检验)、组内治疗前后症状比较(两配对样本t检验)、治疗后组间症候疗效比较(协方差分析);
6、安全性评价治疗前后检查体温,心率,血压,呼吸;血、尿、便常规检查;肝、肾功能检查;心电图检查;
7、统计分析采用统计软件SPSS13.0进行统计学处理和分析。实验数据计量资料采用均数加减标准差((?)±S)表示。治疗前后剂量资料比较采用两配对样本t检验;两组间计量资料比较采用两独立样本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作为差异有显著性差异的界限。
结果
1、一般资料两组在性别(χ~2=0.213,P=0.646)、年龄(F=1.798,P=0.184)、病程(t=1.031,P=0.306),方面比较无显著性差异(P均＞0.05),具有可比性;
2、症候疗效评价两组对“睡眠时间”的疗效对照(Z=-0.633,P=0.527),观察两组对“入睡困难”的疗效对照(Z=-1.716,P=0.086),观察两组对“多梦”的疗效对照(Z=-1.115,P=0.265),均无显著性差异;观察两组对“易醒”症状的疗效对照(Z=-2.285,P=0.022),对“困倦乏力”症状的疗效对照(Z=-2.856,P=0.004),均有显著性差异;
3、整体疗效评价两组治疗对失眠症整体疗效的比较(Z=-2.130,P=0.033),有显著性差异,治疗组总体疗效优于对照组;
4、多导睡眠图(PSG)疗效评价
治疗前两组间,对“总睡眠时间”疗效比较(t=-0.153,P=0.879);“睡眠潜伏时间”疗效比较(t=-0.400,P=0.691);“觉醒次数”疗效比较(t=-0.733,P=0.467);“浅睡眠时间”疗效比较(t=0.290,P=0.773);“深睡眠时间”疗效比较(t=.1.021,P=0.312);“REM睡眠时间”疗效比较(t=.0.425,P=0.673),均无显著性差异。说明两组具有可比性;
治疗前后,治疗组内各症状比较结果:对“总睡眠时间”疗效比较(t=-3.534,P=0.002),“觉醒次数”疗效比较(t=2.753,P=0.011),“深睡眠时间”疗效比较(t=-4.853,P=0.000),“REM睡眠时间”疗效比较(t=-4.596,P=0.000),均有显著性差异;对“睡眠潜伏时间”疗效比较(t=1.572,P=0.129),“浅睡眠时间”疗效比较(t=0.409,P=0.686),无显著性差异。说明治疗组具有“增加总睡眠时间、深睡眠时间、REM睡眠时间,减少觉醒次数”的效果;
对照组内各症状比较:对“总睡眠时间”疗效比较(t=-2.757,P=0.011),“睡眠潜伏时间”疗效比较(t=2.324,P=0.029),“浅睡眠时间”疗效比较(t=-2.396,P=0.025),均有显著性差异;“觉醒次数”疗效比较(t=1.821,P=0.081),“深睡眠时间”疗效比较(t=-1.846,P=0.077),“REM睡眠时间”疗效比较(t=-1.028,P=0.314),均无显著性差异。说明对照组具有“增加总睡眠时间、缩短睡眠潜伏期、增加浅睡眠时间”的效果;
治疗后两组间疗效比较,对“总睡眠时间”疗效比较(F=0.394,P=0.533),对“睡眠潜伏时间”疗效比较(F=1.633,P=0.208),两组对“觉醒次数”的疗效比较(F=1.053,P=0.310),均无显著性差异;对“浅睡眠时间”的疗效(F=9.042,P=0.004),对“深睡眠时间”的疗效比较(F=4.887,P=0.032),对“REM睡眠时间”的疗效比较(F=8.441,P=0.006),均有显著性差异;说明治疗组增加深睡眠时间、REM睡眠时间的疗效优于对照组;对照组增加浅睡眠时间疗效优于治疗组。
结论
1、安神方治疗对改善“易醒,困倦乏力”症状的疗效优于对照组,治疗失眠症的整体疗效优于对照组;
2、安神方对“总睡眠时间、觉醒次数、深睡眠时间、REM睡眠时间”症状的改善,具有显著疗效;与对照组比较,改善“深睡眠时间、REM睡眠时间”的疗效显著。其对睡眠结构的调整主要是增加了“深睡眠时间”及“REM睡眠时间”;致使“总睡眠时间”增加;
3、安全性指标观察各项检查指标无异常。
第二章安神方治疗失眠症的相关机制研究
一、安神方对失眠造模大鼠中枢氨基酸类神经递质的影响
目的观察安神方对大鼠中枢氨基酸类神经递质含量的影响,探讨其治疗失眠症的机理。
方法
1、实验动物与分组清洁级Wistar大鼠60只,体重(200±20g),南方医院实验动物中心提供,许可证号:(粤)2006-0015。按随机方法分成高剂量组、中、低剂量组和生理盐水对照组,共4组;
2、给药高剂量组大鼠每天按40g.Kg~(-1)灌胃,中剂量组大鼠每天按20g.Kg~(-1)灌胃,低剂量组大鼠每天按10g.Kg~(-1)灌胃,生理盐水组给予等量生理盐水灌胃。失眠造模同时连续给药15天;
3、失眠大鼠模型制备方法本实验采用‘限制运动法'制作大鼠失眠模型:将大鼠置于自制的大鼠运动限制仪内,通过移动插片控制大鼠的活动空间。每日束缚制动1次,第一次4h,其后每次增加1h,连续15d完成。大鼠在制动期间禁食、禁水;
4、中枢氨基酸类神经递质检测方法处死并分离大鼠海马及下丘脑。称重,玻璃匀浆器匀浆化;离心取上清液,采用“高效液相色谱荧光检测法”检测中枢氨基酸类神经递质;
5、仪器和试剂
仪器及试剂:DIONEX Summit高效液相色谱仪(美国DIONEX公司),Chromeleon色谱工作站(美国DIONEX公司),RF2000荧光检测器(美国DIONEX公司),PHS-2C精密酸度计(上海理达仪器厂),冷冻离心机(Avanti 30),电子天平(BA 110S)。谷氨酸(Glu)、γ-氨基丁酸(GABA)、OPA(内含2-ME)均购自美国SIGMA公司,批号分别为43907190、026K07361、116K5005;乙腈购自美国TEDIA公司;其余试剂均为市售分析纯;
6、实验数据计量资料采用((?)±S)表示;采用统计软件SPSS13.0进行统计学描述和分析。多组间比较采用单因素方差分析(One-Way ANOVA)检验;方差齐时采用LSD方法检验;方差不齐时采用非参数Kruskal-Wallis检验。所有统计数据均以P＜0.05作为差异有显著性的界限。
结果
1、海马γ-氨基丁酸(GABA)含量高剂量组与生理盐水组比较(P=0.002),中剂量组与生理盐水组比较(P=0.017),低剂量组与生理盐水组比较(P=0.045),均有显著性差异,治疗组GABA含量高于对照组;
2、海马谷氨酸(GLU)含量:高剂量组与生理盐水组比较(P=0.011),中剂量组与生理盐水组比较(P=0.045),均有显著性差异,治疗组GLU含量低于对照组;低剂量组与生理盐水组比较(P=0.566),无显著性差异;
3、下丘脑γ-氨基丁酸(GABA)含量:高剂量组与生理盐水组比较(P=0.001),中剂量组与生理盐水组比较(P=0.028),均有显著性差异,治疗组GABA含量高于对照组;低剂量组与生理盐水组比较(P=0.166),无显著性差异;
4、下丘脑谷氨酸(GLU)含量:高剂量组与生理盐水组比较(P=0.015),有显著性差异,治疗组GLU含量低于对照组;中剂量组与生理盐水组比较(P=0.103),无显著性差异;低剂量组与生理盐水组比较(P=0.798),无显著性差异。
结论安神方治疗组能增加大鼠海马、下丘脑中γ-氨基丁酸(GABA)含量,降低谷氨酸(GLU)含量;调节中枢氨基酸类神经递质是安神方治疗失眠症的机制之一。
二、安神方对睡眠剥夺大鼠血清皮质醇的影响
目的观察安神方对应激条件下动物的焦虑症状的调节作用。
方法
1、动物及分组清洁级Wistar大鼠30只,8周龄,雄性,体重(300±30g),南方医院实验动物中心提供,许可证号:(粤)2006-0015。随机方法分为2组,每组15只;
2、试验药物中药安神方,由南方医院中药房提供。治疗组大鼠每天按20g.Kg~(-1)灌胃(剂量按成人换算),生理盐水组给予等量生理盐水;每天灌胃1次;
3、睡眠剥夺大鼠模型制备采用“改良多平台睡眠剥夺法”进行睡眠剥夺(sleep deprivation,SD)。室温保持20-25℃,正常昼夜光照,每日更换睡眠剥夺用水,保持自由进食条件;
4、试验步骤:持续睡眠剥夺72h,两组分别在睡眠剥夺(SD)Oh,24h,48h和72h时,记录各组动物体重,并采血检测血清皮质醇。血清皮质醇测定方法:于早晨7时经尾静脉取血0.5ml,置离心管中以3000r/min离心10min,取上清液保存于低温冰箱(-60℃)待检测;
5、资料整理与统计学方法:采用统计软件SPSS13.0进行统计学处理和分析。治疗前后指标差值采用协方差分析(analysis of covariance,ANCOVA),不同时间点数据变化采用重复测量数据的方差分析(ANOVA for the repeated measures)和独立样本t检验进行比较(Independent-Samples T Test)。所有统计数据分析均以P＜0.05作为差异有显著性的界限。
结果
1、两组治疗前后体重变化比较(F=79.682,P=0.000),有显著性差异;
2、第1,2天治疗组和对照组的体重比较无显著性差异(第1天:t=-0.429,P=0.671;第2天:t=1.835,P=7.077),而在第3,4天两个时间点上,两组的体重比较均有显著性差异(第3天:t=3.675,P=0.001;第4天:t=5.675,P=0.00)。治疗组和对照组体重在不同时间点之间差异有显著性差异(F=460.01,P=0.000)。分组与时间之间存在交互作用(F=70.22,P=0.000);
3、两组治疗前后皮质醇含量比较(F=70.601,P=0.000),有显著性差异;
4、第1,2天治疗组和对照组的血清皮质醇比较无显著性差异(第1天:t=-0.372,P=0.712;第2天:t=-0.699,P=0.490),而在第3,4天两个时间点上,两组的血清皮质醇比较均有显著性差异(第3天:t=-3.247,P=0.003;第4天:t=-4.770,P=0.000)。治疗组和对照组血清皮质醇在不同时间点之间差异有显著性差异(F=128.94,P=0.000)。分组与时间之间存在交互作用(F=29.846,P=0.000)。
结论
1、治疗组体重减轻速度慢于生理盐水对照组;
2、治疗组血清皮质醇含量增加的速度慢于生理盐水对照组;
3、安神方治疗可以减轻睡眠剥夺大鼠的焦虑症状。
Background
since the ancient times,the humanity on has had the strong interest to the sleep and the dream.The humanity has not stopped regarding the sleep and the dream inquisition since Floyd's "Dream Analysis.As an important constituent,sleep play a important role not only in maintaining human's physiology healthy,but also psychological is.Normal sleep maintains the abundant energy physical strength to the human body,significance of sleep in relation to maintenance nervous system,the endocrine system,and the immune system function normal.
In recent years,anxious and depression from accelerated society transformand and people's life rhythm,made the insomnia occur frequently.Insomnia is a kind of sleep barrier synthesis that something wrong in the onset and maintenance of sleep, far from satisfying the individual physiological needs,affecting the patient's life(for example the daytime sleepy asthenia,remembrance breakdown and so on).The modern medicine discovered the occurrence and the maintenance of sleep had a correlated with not only the suprachiasmatic nucleus,hypothalamus,ascending reticular activating system and apophysis cerebri,but also acetylcholin(ACH), noradrenalin(NE),dihydroxyphenyl ethylamine(DA),the single-amines neurotransmitt e,the amino acid neurotransmitte as well as melatonin,prostaglandin,interleukin.
At present,the main drug to the insomnia is still BZD sedative hypnotics,which bring adverse reaction and drug dependence as well as treat insomnia.The differentiation of Traditional Chinese medicine,adjusts the human performance condition,achieves the improvement of sleep,has brought the gospel for the insomnia patient.Famous physician Chen Baotian of traditional Chinese medicine created "AnShenfang" through reading the Chinese medicine classics "Shanghanlun",and confirmed curatively for a long time in clinical work.simultaneously improved the patient's anxious and despondent.on the base of clinical observation foundation,we designs this experiment to observate the the therapeutic effect of the AnShenfang systematically,and further discusses the mechanism of AnShenfang through the animal experimentation,provides the reliable theory and the clinical basis for the development of AnShenfang.
chapter 1.AnShenfang treats the insomnia clinical observation
Objective To objectively estimate the therapical effect of "AnShenfang" in treating insomnia of Liver-qi and Changes into Liver-fire through the clinical overall curative effect as well as the Polysomnogram(PSG).
Methods
1.Clinical curative effect observation:80 cases are selected and randomly devided into two groups:the AnShenfang group and the control group;
2.According to level of Chinese medicine symptom graduation foundation, compared improvement of main symptom(sleep time,difficulty falling asleep,easily awakening,dreaminess,fatigue),useing the non-parametric test;
3.According to grades around the treatment,calculate "the curative effect index", evaluates rank of improvement in each patient,appraises the total curative effect;
4.Evaluating the therapical effect though Polysonmogram(PSG),records each data(total sleep time,sleep latency time,awareness times,shallow sleep time,deep sleep time,REM sleep time).Objectively appraisal curative effect.
Rusults
1.Clinical observation:there was no significant differences in gender,age and course of disease in two groups(P>0.05),the two groups are comparable;
2.Symptom curative effect appraises:there was no significant differences in "sleep time(Z=-0.633,P=0.527),difficulty falling asleep(Z=-1.716,P=0.086), dreaminess(Z=-1.115,P=0.265)" between two groups;there was significant difference in "easily awakening(Z=-2.285,P=0.022),fatigue(Z=-2.856,P= 0.004) "between two groups;
3.The total curative effect appraises:there was significant difference in total curative effect(Z=-2.130,P=0.033),the treatment group total curative effect surpasses the control group;
4.The polosomnography(PSG) curative effect appraisal:
Before treatment,there was no significant difference in total sleep time compares(t=-0.153,P=0.879);sleep latency time compares(t=-0.400,P=0.691); awareness times compares(t=-0.733,P=0.467);shallow sleep time compares (t=0.290,P=0.773);the deep sleep time compares(t=-1.021,P=0.312);the REM sleep time compares(t=-0.425,P=0.673)between two groups.Two groups are comparable;
Around treatment,comparison in treatment group:there were significant difference in the total sleep time compares(t=-3.534,P=0.002),awareness times compare(t=2.753,P=0.011),the deep sleep time compares(t=-4.853,P=0.000), REM sleep time compares(t=-4.596,P=0.000),there was no significance difference Regarding the sleep latency time compares(t=1.572,P=0.129),and the shallow sleep time compares(t=0.409,P=0.686).Ttreatment group has effect on"increasing the total sleep time,the deep sleep time,the REM sleep time,reduces awareness times;
Symptoms comparison in control group:there were significant differences in the total sleep time compares(t=-2.757,P=0.011),the sleep latency time compares (t=2.324,P=0.029),the shallow sleep time compares(t=-2.396,P=0.025);there was no significant differences in awaken times compares(t=1.821,P=0.081),deep sleep time compares(t=-1.846,P=0.077),REM sleep time compares(t=-1.028,P=0.314), Control group has effect on"the increase sleep total time,the reduce sleep latency time,increases the shallow sleep time";
After the treatment,there were significant differences in the shallow sleep time compares(F=9.042,P=0.004),the deep sleep time compares(F=4.887,P=0.032),the REM sleep time compare(F=8.441,P=0.006) between two groups.there was no in the total sleep time compare(F=0.394,P=0.533),the sleep latency time compare (F=1.633,P=0.208),awareness times compare(F=1.053,P=0.310),increaseing the deep sleep time,the REM sleep time curative effect in treatment group surpasses the control group;The control group increasing the shallow sleep time curative effect surpass the treatment group.
Conclusion
1.Curative effect observation:the effect of "easily awakening,fatigue" in the treatment group surpasses the control group's,and the overall curative effect of treat group surpass the control group;
2.Polysomnogram(PSG) observation:The AnShenfang has significant effect on "the total sleep time,awareness times,the deep sleep time,the REM sleep time", there were significant differences in the "deep sleep time,the REM sleep time" Compared with the control group.The treat group mainly increased "total sleep time"through the increasing of"the deep sleep time" and "the REM sleep time";
3.The treatment security,each inspection target does not have exceptionally.
Chapter 2 laboratory explore of "AnShenfang" treat insomnia
1.The Effect of "AnShenFang"on Central Neurotransmitters of insomnic Rats
Objective Observe the effect of "AnShenFang"on Central Neurotransmitters of insomnic Rats,discusses the mechanism to treat the insomnia.
Methods
1.The experimental animal and group:60 Wistar rats,the body weight (200±20g),south the hospital experimental animal center provides,permit number: (Guangdong) 2006-0015.were randomly divided into the high dose group,middle dose group,lowdose group and the saline control group;
2.Administration:40g.Kg~(-1) for the high dose group.20g.Kg~(-1) for the middle dose group,and 10g.Kg~(-1) for the low dose group and the saline for the control group;meanwhile,made the insomnia model for 15 day;
3.insomnia rats model preparation:making insomnia rats model through limiting rats'movement:rats were Put in self-maderats movement limit meter,controls the rat's active space through the motion film threading,applies the brake 1 time each day, first time 4h,increases 1h each time after that,sustaining for 15d,diet and water were prohibited in this period;
4.examination of the central nervous:separate both hippocampus and hypothalamus of the rat.weighting,glass homogenizer refining;centrifugalization, quantitating through the high performance liquid chromatogram(HPLC);
5.Uses statistical software SPSS13.0 to 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.
Rusults
1.Gamma-amino-butyric acid(GABA) content in hippocampus:there were significant differences among the high dose group and saline control group compare (P=0.002),the middle dose group and saline control group compare(P=0.017),the low dose group and saline control group compare(P=0.045);
2.Glutanic acid(GLU) content in hippocampus:there were significant differences among the high dose group and the saline control group compare (P=0.011),the middle dose group and the saline control group compare(P=0.045); there was no significant difference between the low dose group and saline control group compare(P=0.566);
3.Gamma-amino-butyric acid(GABA) content in hypothalamus:there were significant differences among the high dose group and saline control group compare (P=0.001),the middle dose group and the saline control group compare (P=0.028);there was no significant difference between the low dose group and the saline control group compare(P=0.166);
4.Glutanic acid(GLU) content in hypothalamus:there was significant difference between the high dose group and the saline control group compare (P=0.015);there were no significant differences between the middle dose group and the saline control group compare(P=0.103),as well as the low dose group and the saline control group compare(P=0.798).
Conclusion The AnShenfang can elevate the Gamma-amino-butyric acid (GABA) content,reduces the glutanic acid(GLU) content both hippocampus and hypothalamus of the rats.
2.The Effect of "AnShenfang"on serum cortisol of Sleep Deprivated Rats
Objective To observe the effect of "AnShenfang" on Serum cortisol of Sleep Deprivated Rats.
Methods
1.The experimental animal and group:30 rats were randomized divided into 2 groups:treat group and normal control group,each group had 15 animals.Treat group were fed by "AnShenFang"and normal control group were fed by normal saline for 3 days,then sleep deprivated for 72 hours,test the body weight and content of serum cortisol on sleep deprivation 0h,24h,48h and 72h;
2.Administration "AnShenfang"20g.Kg~(-1) for the treat group,and the saline for the control group;
3.The rats sleep deprives model preparation:use"the multi-platform sleep deprivation methord" prepare sleep deprivation model(sleep deprivation,SD).The room temperature maintenancing 20-25℃,replacing the the water used every day, maintenancing free feed condition;
4.Testing methord:sleep depriving for 72h Continuesly,records each group of animal body weight,and detects blood serum cortisol on 0h,24h,48h and 72h.Blood serum cortisol determination method:on7 AM draws blood 0.5ml from the tail vein, centrifuge at 3000r/min for 10min,preserve serumin the low temperature refrigerator for detect(-60℃);
5.Uses statistical software SPSS13.0 to detail the data.use the analysis of covariance(ANCOVA)around test;uee the ANOVA for the repeated measures and Independent-Samples T Test to test the data on the different time All statistical analysis has the significance boundary as P<0.05.
Rusults
1.There were significant differences in the body weight of the two groups compare(F=79.682,P=0.000);
2.There was no significant difference on body weight at the 1,2nd day between the treat group and the control group's compares(1st day:t=-0.429,P=0.671;2nd days:t=1.835,P=7.077),but there were significant differences on bodu weight between weo group on the 3,4th day(3rd days:t=3.675,P=0.001;4th days:t=5.675, P=0.00).there was significant difference on body weight on different time(F=460.01, P=0.000).Between the grouping and the time has the correlation(F=70.22,P=0.000);
3.There were significant differences in the serum cortisol of the two groups compare(F=70.601,P=0.000);
4.There was no significant difference on content of serum cortisol at the 1,2nd day between the treat group and the control group's compares(1st day: t=-0.372,P=0.712;2nd days:t=-0.699,P=0.490),but there were significant differences on content of serum cortisol between two groups on the 3,4th day(3rd days:t=3.675, P=0.001;4th days:t=5.675,P=0.00).there was significant difference on content of serum cortisol on different time(F=460.01,P=0.000).Between the grouping and the time has the correlation(F=70.22,P=0.000).
Conclusion
1.The speed of body weight of Treat group was slow compared with saline control group;
2.The speed of content of serum cortisol of treat group was slower than compared with saline control group;
3.Anshenfang granules can elivate the rat's anxious symptom which reduce the sleep deprive.
引文
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