调肝方药对慢性束缚应激大鼠神经内分泌免疫功能的影响

七情致病理论及其防治是中医病因病机学中的一个重要内容,但目前相关研究较少,现代医学中的心理应激理论与中医七情学说在认识上存在着许多共同点,从应激角度出发,结合中医"怒伤肝"的理论,以束缚制动作为应激原,复制慢性激怒应激反应大鼠模型,并探讨调肝方药(TGFY)对应激大鼠神经内分泌免疫功能的影响。1 材料与方法1.1 主要试剂 细胞培养液RPMI-1640(Gibco产品),刀豆素A(ConA,Sigma产品),3H-TdR(中国科学院原子能研究所产品),莨菪葶(9．61μg,Sigma产品),辣根过氧化物酶(HRP,10μg,Sigma产品),十四酰佛波乙酸酯(PMA,100ng,Sigma产品)。125I-CRH、125I-ACTH、125I-CORT放免检测药盒(美国DPC产品)。1.2 大鼠应激模型制作及分组处理 雄性Wistar大鼠,体重250～300 g,分为正常对照组10只(正常饮食,不给任何刺激);慢性束缚应激组1O只(给予束缚刺激,正常饮食,不给中药治疗);调肝方药组10只(给予束缚刺激的同时灌胃调肝方药)。慢性束缚应激反应大鼠模型的制作取雄性Wistar大鼠20只,体重250～300 g,每2只一笼,每日给予黑暗和光照各12 h,20～22℃,同时给予每日1次成对双后肢的立式束缚,以引起明显的激怒应激反应(粗叫、站立对峙、扭打嘶咬),首次30min,以后隔日增加10min,共2 w。普通饲料,自由饮水。于实验第13天和第14天在动物清醒、安静状态下断头处死取材。1.3 药物组成剂量及给药方法 所用中药均由本校第一附属医院药房提供的纯正药材。调肝方药组成及剂量：柴胡15g、白芍15 g,香附10 g,山萸肉15 g,郁金10 g,枳壳10g。将上述药物制成100％水煎剂,每毫升含生药1 g,给予大鼠1 ml／100 g体重灌胃,每日2次,共2 w。对照组及应激组灌服等量生理盐水。     

自主跑轮运动对慢性不可预见性轻度应激大鼠抑郁样行为及神经内分泌因子近日节律的影响

目的:探讨8周自主跑轮运动对慢性不可预见性轻度应激(CUMS)大鼠抑郁样行为及外周神经内分泌因子生物节律表达的影响,从时间生物学方面探讨运动抗抑郁、促进脑康复的作用机制。方法:雄性SD大鼠90只,随机分为对照(control)组、模型(model)组及模型运动(model+exercise)组。采用CUMS方式建立抑郁模型,模型运动组前5周自愿跑轮,从第6周开始,附加21 d慢性复合应激,模型组于同一时点造模21 d。采用糖水偏好实验、旷场行为及高架十字迷宫系统观测大鼠焦虑情绪的行为学变化。行为学检测结束后第2天,连续24 h分6个时点(ZT1、ZT5、ZT9、ZT13、ZT17、ZT21)处死动物取血,每个时点处死5只大鼠,放射免疫法测定上述6个时点血清促肾上腺皮质激素(ACTH)含量,ELISA检测6个相同时点血清皮质酮(CORT)、褪黑素(MT)及血管活性肠肽(VIP)的含量,采用单一余弦法拟合曲线比较慢性应激大鼠各指标的节律周期、振幅、峰相位、中值的变化特点以及运动对其的影响。结果:与对照组相比,21 d的CUMS使模型组大鼠体重增加量明显减少(P0.01),糖水消耗量明显降低(P0.01);旷场实验中爬格数、直立次数及中央格停留时间均显著下降,而粪粒数明显增多(P0.01);高架十字迷宫中开放臂停留时间的百分比和开放臂进入次数的百分比明显降低(P0.01)。运动对CUMS诱导的大鼠抑郁样行为有明显的改善作用。血浆近日节律输出肽及相关激素方面,慢性应激至抑郁样行为充分表达后,模型组大鼠血浆ACTH、CORT的相位与对照组完全相反,表现为时相的大幅度提前、含量波动幅度减小、昼夜分泌节律紊乱;MT的24 h分泌节律完全丧失且整体水平下降,表达量显著低于对照组;VIP虽仍存在24 h节律,但振幅明显低于对照组,峰相位也延迟6 h,表达量显著高于对照组。8周跑轮运动能显著改善应激大鼠的抑郁样行为及激素分泌量的异常,使血浆ACTH、MT、CORT和VIP的近日节律恢复正常。结论:运动可拮抗CUMS致大鼠的抑郁样行为,其作用机制可能与其在一定程度上能够调整外周神经内分泌激素的昼夜节律性和改变激素的分泌量有关。     

影响实验动物慢性心理应激反应的因素

动物慢性心理应激是研究焦虑症、抑郁症以及情绪障碍等人类的精神疾病比较常用的方法。在动物实验中,慢性应激对动物的影响是多方面、持久的并且比较复杂,包括神经内分泌、免疫系统以及行为学的变化等,这些变化的原因可以分为应激源因素如应激源类型、应激强度和应激时间等以及非应激源因素如实验动物年龄、性别、种属等。本文旨在探讨这两种因素在慢性心理应激中的影响,以便对将来的研究提供参考和借鉴。     

跳伞应激对空降兵新兵神经内分泌的影响

近年来神经内分泌研究成为预防医学和军事医学的优先发展领域和研究前沿。其研究对提高空降兵部队的心理健康水平和紧急状态下的应对能力 ,最大限度减低非战斗减员的发生率具有重要意义。国外跳伞人员 (非伞兵 )神经内分泌学研究偏重于与应激有关的下丘脑—垂体—肾上腺轴系统 (皮质醇、ＡＣＴＨ等 )及内源性阿片肽的研究[1-6] ,对其它系统及神经肽的变化研究甚少。本研究就空降兵新兵群体在跳伞应激状态下精氨酸血管加压素 (ＡＶＰ)、神经肽Ｙ(ＮＰＹ)、降钙素基因相关肽 (ＣＧＲＰ)、神经降压素 (ＮＴ)、血管活性肠肽 (ＶＩＰ)的变化规律…     

慢性复合应激对成年大鼠海马NeuroD表达的影响

本实验通过观察慢性复合应激对成年大鼠海马NeuroD表达的影响,探讨慢性复合应激与海马神经发生的关系。将成年雄性大鼠32只随机分为慢性复合应激组(简称应激组)和正常对照组(简称对照组)。应激组动物每天不规律交替暴露于四种复合应激原中,共持续6周。然后运用免疫组织化学染色、Western-blot和RT-PCR技术分别观察海马内NeuroD阳性神经元数量、NeuroD蛋白水平和核酸水平的变化。结果显示:慢性复合应激组动物海马齿状回NeuroD阳性神经元数量明显增多(P<0.05);海马NeuroD蛋白的表达明显增强(P<0.05);海马NeuroD mRNA水平明显上调(P<0.05)。表明慢性复合应激可引起海马NeuroD阳性神经元数量增多和NeuroD表达水平升高,提示慢性复合应激可能促进大鼠海马的神经发生。     

慢性复合应激对大鼠学习和记忆功能及齿状回神经前体细胞增殖的影响

为研究慢性复合应激对成年雄性大鼠学习和记忆功能及齿状回(DG)神经前体细胞增殖的影响,将成年雄性Wistar大鼠随机分为对照组和应激组,应激组动物每天交替暴露于复合应激原中,持续6周。应激结束后,用Morris水迷宫测试大鼠空间学习记忆成绩。同时,采用BrdU标记分裂细胞方法,观察比较各组大鼠DG内BrdU阳性细胞数的变化和差异。结果显示:应激组动物的学习与记忆成绩均优于对照组(P<0.05);与对照组相比,应激组大鼠DG内BrdU阳性细胞数量显著增加(P<0.05)。上述结果表明:慢性复合应激导致大鼠的学习与记忆能力加强,DG内BrdU阳性细胞增多,提示神经细胞数量增加可能是应激所引起的大鼠学习记忆能力增强的原因之一。     

氯化锂对染铅大鼠神经行为的影响

目的探讨不同浓度氯化锂对染铅大鼠神经行为的影响。方法将大鼠随机分为对照组、染铅组、四个氯化锂组和四个铅+氯化锂组,分别给予普通饲料和含氯化锂(3,30,300和3000mg/kg)的饲料喂养,染铅组和四个铅+氯化锂组饮用体积分数0.2%PbAc2的单蒸水,喂养60d,通过测量大鼠体重和Y-迷宫实验,观察各组大鼠之间学习记忆能力的差别;采用黄递酶(NADPH-d)组织化学染色方法观察各组大鼠海马一氧化氮合酶(NOS)阳性细胞数的差别。结果氯化锂(3,30mg/kg)组大鼠体重增长多于对照组,Y-迷宫实验学会次数均少于对照组,差异均有显著性(P<0.05);氯化锂(300、3000mg/kg)组、染铅组体重增长少于对照组,Y-迷宫实验学会次数均多于对照组,差异均有显著性(P<0.05);与染铅组相比,铅+氯化锂(3,30,300mg/kg)组大鼠体重增量增加,Y-迷宫实验学会次数减少,差异有显著性(P<0.05)。NADPH-d组织化学染色结果示:氯化锂(3,30mg/kg)组大鼠海马一氧化氮合酶(NOS)阳性细胞数明显多于对照组(P<0.05);氯化锂(300、3000mg/kg)组、染铅组则少于正常大鼠(P<0.05);铅+氯化锂(3,30,300,3000mg/kg)组大鼠NOS阳性细胞数均多于染铅组(P<0.05)。结论较低浓度的氯化锂能促进正常大鼠学习记忆能力的提高,而较高浓度的锂则可能损害大鼠的学习记忆能力;铅暴露大鼠学     

心理社会因素与银屑病患者神经内分泌免疫指标的关系

目的:从神经内分泌免疫网络调节来探讨心理社会应激影响银屑病的可能中介机制.方法:运用临床心理测定工具(TABPQ,TAS,SAS,SDS)测定40例银屑病患者及38例正常对照组在病前遭遇的人格特征、患病后的心身健康状况;用放射免疫分析方法分别测定40例银屑病患者的神经肽Y(NPY)、白介素6(IL-6)、白介素8(IL-8)、皮质醇(Cortisol)的含量,并与38例健康对照组进行对照研究;将患者的心理社会因素与生物学指标进行Pearson相关分析.结果:1.病例组A型行为评分、TAS总分、抑郁及焦虑分值均显著高于对照组.2.病例组的IL-6、IL-8均显著高于对照组(P＜0.01),NPY和Cor的含量高于对照组(P＜0.05).3.通过相关分析发现患者的抑郁焦虑分、TAS总分与IL-6、IL-8、NPY、Cortisol有显著正相关性,TH CH分与NPY、IL-6有显著正相关性,TH CH分与IL-8、Cor没有显著相关性.结论:心理社会指标与银屑病患者的某些神经内分泌免疫指标有相关性.     

异丙基肾上腺素诱导大鼠慢性心力衰竭神经内分泌与细胞因子的改变

目的 建立异丙基肾上腺素(Isopreterenol,ISO)诱导SD大鼠慢性心力衰竭(CHF)模型,观察神经内分泌与细胞因子的变化.方法 用ISO(170 mg*kg-1)对SD大鼠皮下注射2次,建立大鼠CHF模型,在造模后18周进行心脏超声检测左室舒张末径(LVEDD)、左室收缩末径(LVESD)、射血分数(LVEF)、短轴缩短率(FS),取血检测去甲肾上腺素(NE)、血管紧张素Ⅱ(ANGⅡ)、醛固酮(ALD)、肿瘤坏死因子α(TNF-α),并取心肌观察病理形态学特征. 结果 与正常对照组相比,模型组大鼠LVEF、FS下降,LVEDD、LVESD扩大(P＜0.01),左心室重量指数明显增加(P＜0.01),NE、ANGⅡ、ALD、TNF-α显著升高(P＜0.01).光学显微镜结果 显示,ISO组大鼠心肌细胞肥大,部分心肌细胞有嗜酸变性和/或坏死,心肌间质结缔组织增生. 结论 应用大剂量ISO诱导大鼠慢性心力衰竭出现神经内分泌、细胞因子激活.     

恐惧应激对大鼠心血管系统及一般行为的影响

目的 观察恐惧应激对大鼠血压、心率及一般行为的影响.方法 采用条件恐惧应激模型,将64只成年雄性Wistar大鼠随机分为恐惧应激模型组、对照组.通过对大鼠恐惧行为的观察、自发活动试验的测定,动态观察两组大鼠在不同恐惧应激时期行为学方面的变化;通过无创尾动脉测压法,观察不同恐惧应激时程大鼠血压及心率的变化;采用放射免疫学...     

Effects of alcohol exposure during development on social behavior in rats

In addition to the cognitive deficits associated with fetal alcohol syndrome (FAS), clinical and animal studies indicate that alcohol exposure might also have detrimental effects on social behavior. In a rat model of FAS, experimental rats were given alcohol from gestational day (GD) 1 to 22 and from postnatal day (PD) 2 to 10, a period roughly equivalent to all three trimesters in humans. Control groups consisted of rats exposed to the administration procedures but not to alcohol and nontreated rats. At 30 days of age, rats were tested for social behavior in an alley maze that contained its cagemate in the goal box. After varying periods of isolation, the animals' latencies to reach the goal box and their social behaviors once inside the goal box were recorded. Alcohol-exposed animals ran faster than control rats to the occupied goal box regardless of the amount of isolation. The alcohol-exposed animals also exhibited aberrant social interactions with their cagemate once inside the goal box compared to one or both of the control groups. Specifically, the alcohol-exposed animals showed greater amounts of anogenital sniffing, chasing, hopping and darting, and retrieving and lesser amounts of pinning and biting compared to one or both of the control groups. The alcohol-induced change in anogenital sniffing varied over increasing amounts of isolation compared to both control groups, but the alterations in the other behaviors did not. It is argued that the altered social behavior of alcohol-exposed animals is not the result of changes in the animals' motivational state or social learning and may be the result of an increased responsiveness to social stimuli.     

睾酮对慢性心力衰竭雄性大鼠神经内分泌及细胞因子系统的影响

目的： 探讨睾酮对慢性心力衰竭雄性大鼠神经内分泌激素以及炎性细胞因子的影响。方法: 74只雄性SD大鼠分为①正常对照组(control)；②心力衰竭组(CHF)；③睾丸切除+心力衰竭组(ORX+CHF)；④睾丸切除+心力衰竭+睾酮替代治疗组(ORX+CHF+T)。应用超声学检测大鼠心功能，放射免疫法检测血浆中睾酮（T）、ANP和细胞因子（TNF-α、ET-1）水平。结果: ⑴ CHF组大鼠血浆T浓度明显低于control组（P＜0.01）。心力衰竭大鼠血浆T水平与左室短轴缩短率（FS％）呈正相关（R＝0.54,P＜0.01）。⑵ 心力衰竭大鼠的血浆ANP水平较正常降低。心力衰竭大鼠血浆TNF-α、ET-1浓度明显高于正常，相比ORX+CHF组睾酮替代治疗拮抗了血浆TNF-α的升高(P＜0.05），但对ET-1的影响不明显。结论: 在扩张型心肌病慢性心力衰竭雄性大鼠，血浆雄激素水平与左室收缩功能具有良好的相关性，而体内正常的雄激素水平对神经内分泌及细胞因子系统的异常激活具有有益的调节作用。     

Effects of chronic administration of caffeine and stress on feeding behavior of rats

Anorectic effects of caffeine are controversial in the literature, while stress and obesity are growing problems in our society. Since many stressed people are coffee drinkers, the objective of the present study was to evaluate the effect of stress and chronic administration of caffeine on feeding behavior and body weight in male and female rats. Wistar rats (both males and females) were divided into 3 groups: control (receiving water), caffeine 0.3 g/L and caffeine 1.0 g/L (in the drinking water). These groups were subdivided into non-stressed and stressed (repeated-restraint stress for 40 days). During the entire treatment, chow consumption was monitored and rats were weighed monthly. Afterwards, feeding behavior was evaluated during 3-min trials in food-deprived and ad libitum fed animals and also in repeated exposures, using palatable food (Froot Loops® and Cheetos®). Chronic administration of caffeine did not affect rat chow consumption or body weight gain, but diminished the consumption of both salty (Cheetos®) and sweet (Froot Loops®) palatable food. In the repeated trial tests, stress diminished savory snack consumption in the later exposures [I.S. Racotta, J. Leblanc, D. Richard The effect of caffeine on food intake in rats: involvement of corticotropin-releasing factor and the sympatho-adrenal system. Pharmacol Biochem Behav. 1994, 48:887-892; S.D. Comer, M. Haney, R.W. Foltin, M.W. Fischman Effects of caffeine withdrawal on humans living in a residential laboratory. Exp Clin Psychopharmacol. 1997, 5:399-403; A. Jessen, B. Buemann, S. Toubro, I.M. Skovgaard, A. Astrup The appetite-suppressant effect of nicotine is enhanced by caffeine. Diab Ob Metab. 2005, 7:327-333; J.M. Carney Effects of caffeine, theophylline and theobromine on scheduled controlled responding in rats. Br J Pharmacol. 1982, 75:451-454] and caffeine diminished consumption of both palatable foods (savory and sweet) during the early and later exposures. Most responses to caffeine were stronger in females, and stress exposure influenced the effect. Neither chronic caffeine nor stress affected adrenal weight and plasma corticosterone levels of the rats. These observations suggest that chronic caffeine consumption may have sex-specific effects on palatable food ingestion.     

Aging-related changes in the effects of social isolation on social behavior in rats

Aging is generally associated with cognitive dysfunction and alterations in emotional response. Moreover, in social situations, aging decreases social interaction with unfamiliar individuals, suggesting the decline of social cognition/motivation and a high level of anxiety. Although it is known that isolation housing has various effects on subsequent behavior, including social interaction depending on the age at isolation, the effects of isolation on aged subjects have not been examined. In the present study, we investigated the effects of aging and different periods of isolation housing on social interaction in male F344/N rats. Young (3-4 months old) and aged (24-25 months old) rats were either group-housed or socially isolated for 2 or 4 weeks. The rats were tested with age-matched and group-housed unfamiliar males in a social interaction test, and social (e.g. approach/following and sniffing) and non-social behaviors (e.g. self-grooming and ambulation) were recorded. The results indicated that group-housed aged rats showed less approach/following, sniffing, and ambulation than group-housed young rats. Moreover, in young rats, isolation housing gradually increased approach/following and sniffing depending on the isolation period. In contrast, in aged rats, more prolonged isolation (4 weeks) attenuated the 2-week isolation-induced increase of sniffing behavior and had no effect on approach/following. The present study suggests that aging decreases social investigation and induces high emotional response to a novel social environment, and that the behaviors can be differentially affected by social isolation depending on the age at isolation and the period of isolation.     

Announced reward counteracts the effects of chronic social stress on anticipatory behavior and hippocampal synaptic plasticity in rats

Chronic stress causes insensitivity to rewards (anhedonia) in rats, reflected by the absence of anticipatory behavior for a sucrose-reward, which can be reversed by antidepressant treatment or repeated announced transfer to an enriched cage. It was, however, not clear whether the highly rewarding properties of the enriched cage alone caused this reversal or whether the anticipation of this reward as such had an additional effect. Therefore, the present study compared the consequences of the announcement of a reward to the mere effect of a reward alone with respect to their efficacy to counteract the consequences of chronic stress. Two forms of synaptic plasticity, long-term potentiation and long-term depression were investigated in area CA1 of the hippocampus. This was done in socially stressed rats (induced by defeat and subsequent long-term individual housing), socially stressed rats that received a reward (short-term enriched housing) and socially stressed rats to which this reward was announced by means of a stimulus that was repeatedly paired to the reward. The results were compared to corresponding control rats. We show that announcement of enriched housing appeared to have had an additional effect compared to the enriched housing per se as indicated by a significant higher amount of LTP. In conclusion, announced short-term enriched housing has a high and long-lasting counteracting efficacy on stress-induced alterations of hippocampal synaptic plasticity. This information is important for counteracting the consequences of chronic stress in both human and captive rats.     

慢性缺氧对大鼠行为和海马齿状回DCX阳性细胞的影响

目的:研究慢性低压性缺氧对大鼠行为和海马齿状回DCX阳性细胞的影响。方法:建立SD大鼠慢性低压性缺氧模型,通过开场实验和蔗糖饮水实验观察大鼠行为变化。取脑组织冰冻切片,用免疫荧光检测海马齿回神经元前体细胞标志物微管相关蛋白Doublecortin(DCX)检测海马齿状回DCX阳性细胞的变化。结果:与对照比较,慢性低压性缺氧大鼠体重,开场实验水平运动得分,垂直运动得分降低和蔗糖偏嗜度均明显降低(P0.01)。免疫荧光双标检测显示,慢性低压性缺氧大鼠海马齿状回颗粒下层(SGZ)DCX阳性细胞减少;DCX阳性细胞平均胞体周长(μm):实验组(151.2±16.1)明显大于对照组(132.8±14.5,P0.01);DCX阳性细胞突起平均长度(μm):实验组(21.7±7.3)明显大于对照组(15.2±5.6,P0.01)。结论:慢性低压性缺氧大鼠行为学上可发生明显改变,海马齿状回DCX阳性细胞再生能力减弱,提示缺氧导致该细胞再生损害可能是发生行为学改变的基础。     

Effects of acupuncture on chronic corticosterone-induced depression-like behavior and expression of neuropeptide Y in the rats

Repeated injection of corticosterone (CORT) induces dysregulation in the HPA axis, resulting in depression and anxiety. Many studies have shown that acupuncture, which is widely used for the treatment of stress and mental illness, in East Asian countries, is an effective therapeutic intervention for psychosomatic disorders. We investigated the influence of acupuncture therapy on chronic CORT-induced behavioral responses to the forced swimming test (FST) and elevated plus maze (EPM) and expression of neuropeptide Y (NPY) in the rat brain using immunohistochemistry. Male Sprague-Dawley rats were injected with CORT (40 mg/kg, i.p.) once daily for 19 consecutive days. The dysregulation of HPA axis by external injection of CORT was confirmed by measuring the CORT concentration in plasma and the expression level of CRF in hypothalamus. Acupuncture was performed at the PC6 acupoint for 5 min before CORT injection. Acupuncture significantly reduced depression- and anxiety-like behavior and increased NPY expression in the hypothalamus. These results demonstrated that stimulation of the PC6 acupoint suppresses the symptopathology of the hypoactivated HPA axis in chronic CORT-induced rat model of depression.