摘要
背景:人体处于应激状态时,脑内的一些氨基酸作为神经递质对脑功能和心理行为具有重要的调节作用,传统中药治疗能够调整人体的应激状态。目的:通过观察反复心理应激大鼠下丘脑和海马的谷氨酸、天冬氨酸、γ-氨基丁酸、牛磺氨酸的含量变化,探讨调肝方、健脾方、补肾方及人参总皂苷对其影响。设计:随机分组,对照观察实验。单位:广州中医药大学基础医学院中医基础理论教研室。材料:实验于2002-03/2003-01在广州中医药大学实验动物中心完成。选取Wistar大鼠60只,随机分为6组:正常组,模型组,调肝组,补肾组,健脾组,人参总皂苷组,10只/组。调肝方药组成及剂量:柴胡5g,山栀5g,白芍15g,枸杞子15g,枳壳6g,干地黄18g,石决明30g。肾气丸组成及剂量:干地黄30g,山药15g,山茱萸15g,泽泻10g,茯苓10g,丹皮10g,桂枝4g,附子4g。四君子汤组成及剂量:党参20g,白术15g,茯苓15g,炙甘草6g。方法:①常规煎煮取汁后调肝方药浓缩至含生药1.69g/mL,肾气丸浓缩至含生药1.76g/mL,四君子汤浓缩至含生药1.01g/mL,人参总皂苷配制成水溶液7g/L。正常组及模型组灌服9.0g/L氯化钠注射液2mL,调肝组、补肾组、健脾组及人参总皂苷组均于限制制动刺激前1h相应给予调肝方药、肾气丸煎剂、四君子汤煎剂及人参总皂苷溶液各2mL灌胃。②除正常组外,其余各组均进行心理应激反应模型的复制。将大鼠置于限制制动筒内,通过移动插片而逐步缩小大鼠的活动空间,调节到其不产生强烈反抗的紧张程度,每天限制制动1次,每天限制制动时间不同,第1天为4h,其后每次增加30～60min,连续14d。③将各组大鼠断头取全脑,采用OPA高效液相色谱分析法进行下丘脑和海马中谷氨酸、天冬氨酸、γ-氨基丁酸、谷氨酸含量的检测。主要观察指标:各组大鼠下丘脑与海马中谷氨酸、天冬氨酸、γ-氨基丁酸、牛磺氨酸含量的变化。结果:实验纳入大鼠60只,全部进入结果分析。①各组大鼠下丘脑与海马中谷氨酸含量的变化:与正常组比较,调肝组、健脾组、补肾组、人参总皂苷组下丘脑中含量均明显下降[(21.85±8.19),(15.76±1.80),(14.68±7.91),(21.46±5.45),(13.43±7.68)μmoL/g];与模型组比较,调肝组、健脾组、补肾组、人参总皂苷组海马中含量均明显升高[(11.04±3.65),(11.78±2.17),(18.67±2.98),(20.91±3.96),(17.71±1.83)μmoL/g,P<0.05,P<0.01,P<0.01,P<0.05]。②各组大鼠下丘脑与海马中天冬氨酸含量的变化:与模型组比较,健脾组和人参总皂苷组下丘脑中含量均明显下降[(8.65±1.18),(5.72±1.32),(4.67±1.88)μmoL/g,P<0.01,P<0.01],而健脾组、补肾组、人参总皂苷组海马中含量均明显升高[(2.58±0.87),(3.93±0.49),(4.52±0.98),(3.83±0.41)μmoL/g,P<0.01,P<0.01,P<0.05]。③各组大鼠下丘脑与海马中γ-氨基丁酸含量的变化:与模型组比较,调肝组、健脾组、补肾组、人参总皂苷组下丘脑中含量均明显下降[(20.92±4.96),(15.87±2.90),(13.84±2.63),(14.94±3.98),(10.94±3.68)μmoL/g,P<0.05,P<0.05,P<0.01,P<0.01],而海马中含量均明显升高[(4.12±1.66),(4.18±1.04),(6.67±1.29),(6.11±0.99),(6.37±0.78)μmoL/g,P<0.05,P<0.01,P<0.01,P<0.01]。④各组大鼠下丘脑与海马中牛磺氨酸含量的变化:与模型组比较,调肝组、健脾组、人参总皂苷组下丘脑中含量均明显下降[(10.24±1.72),(7.82±1.14),(8.00±2.05),(6.42±3.17)μmoL/g,P<0.05,P<0.05,P<0.01],而健脾组和人参总皂苷组海马中含量均明显升高[(12.61±3.51),(17.03±2.74),(18.04±2.14)μmoL/g,P<0.01,P<0.01]。结论:调肝方药的中枢作用部位可能主要在下丘脑,可能与下调氨基酸水平有关;而健脾、补肾方药及人参总皂苷的中枢作用部位可能包括海马和下丘脑,主要与上调氨基酸水平有关,通过增强海马对应激的整合作用,从而达到抗应激损伤的目的。
BACKGROUND： When one is in a stress state, some amino acids as neurotransmitter in his brain are of important regulating action to his cerebral functions and his psychic behaviors, and some traditional Chinese drugs can regulate the stress state of the body. OBJECTIVE： To observe the content changes of glutamic acid, aspartic acid, γ-aminobutyric acid and taurine in the hypothalamus and hippocampus of rats under repeatedly psychic stress so as to investigate the effects of tiaogan recipe, jianpi recipe, bushen recipe and ginsenoside on them. DESIGN： A randomized grouping and controlled observation trial. SETTING：Department of Basic Theory of Traditional Chinese Medicine （TCM） of Basic Medical Science College, Guangzhou University of TCM. MATERIALS： The experiment was completed from March 2002 to January 2003 at the Animal Center of Guang, zhou University of TCM. Totally 60 Wistar rats were randomly divided into 6 groups： normal, model, tiaogan, bushen, jianpi and ginsenoside groups, with 10 in each group. Compositions and doses of tiaogan recipe： radix bupleuri 5 g, fructus gardeniae 5 g, radix paeoniae alba 15 g, fructus lycii 15 g, fructus aurantii 6 g, radix rehmanmae 18 g, concha hahohdis 30 g. Compositions and doses of shenqi pill： radix rehmanniae 30 g, rhizoma dioscoreae 15 g, fructus corni 15 g, rhizoma alismatis 10 g, poria 10 g, cortex moutan radicis 10 g, ramulus cinnamomi 4 g, radix aconiti praeparata 4 g. Compositions and doses of sijunzi decoction： radix codonopsis pilosulae 20 g, rhizoma atractylodis macrocephalae 15 g, poria 15 g, radix glycyrrhizae preparata 6 g. METHODS： ① The traditional Chinese medicines were conventionally decocted; and the tiaogan recipe condensed to the liquid containing 1.69 g/mL crude drug, shenqi pill containing 1.76 g/mL crude drug, sijunzi decoction containing 1.01 g/mL crude drug. Ginsenoside was prepared as 7 g/L water solution. The rats in the normal and model groups were by garage given 2 mL of 9.0 g/L sodium chloride injection. The rats in the tiaogan, bushen, jianpi and ginsenoside groups were respectively by garage given 2 mL of tiaogan recipe, shenqi pill, sijunzi decoction and ginsenoside solution 1 hour before immobilization stress. ② Except for rats in the normal group, those in the rest groups were all conducted for establishment of psychic stress reaction model. The rats were put into an immobilization tube, their action space was gradually reduced by using a mobile insertion piece, they were regulated to a nervous state without production of intense revolting, which was done once a day, starting with 4 hours immobilization on the first day, and later on increased by 30-60 minutes per day, for consecutive 14 days. ③ The whole brain of the rats in each group was collected by decapitation, OPA high-performance liquid chromatography was used for assays of the contents of glutamic acid, aspartic acid, γ- aminobutyric acid and taurine in hypothalamus and hippocampus of the rats. MAIN OUTCOME MEASURES： The content changes of glutamic acid, aspartic acid, γ-aminobutyric acid and taurine in the hypothalamus and hippocampus of rats in each group. RESULTS： Totally 60 rats involved all entered into the result analysis. ① The content changes of glutamic acid in the hypothalamus and hippocampus of rats in each group： Compared with normal group, the contents of glutamic acid in hypothalamus of rats in the tiaogan, jianpi, bushen and ginsenoside groups were markedly decreased [（21.85±8.19）, （15.76±1.80）, （14.68±7.91）, （21.46±5AS）, （13.43±7.68） μmoL/g]; compared with model group, the contents in the tiaogan, bushen, jianpi and ginsenoside groups were obviously raised [（11.04±3.65）, （11.78±2.17）, （18.67±2.98）, （20.91±3.96）, （17.71±1.83） μmoL/g, P 〈 0.05, P 〈 0.01, P 〈 0.01, P 〈 0.05].② The content changes of aspartic acid in the hypothalamus and hippocampus of rats in each group： Compared with model group, the contents of aspartic acid in the hypothalamus in＇the fianpi and ginsenoside groups were obviously decreased [（8.65±1.18）, （5.72±1.32）, （4,67±1.88） μmoL/g, P 〈 0.01, P 〈 0.01], while the contents in the hippocampus of rats in the jiartpi, bushen and ginsenoside groups were markedly raised [（2.58 ±0.87）, （3.93±0.49）, （4.52±0.98）, （3.83±0.41） μmoL/g, P 〈 0.01, P 〈 0.01, P 〈 0.05]. ③ The content changes of γ-aminobutyfic acid in the hypothalamus and hippocampus of rats in each group： Compared with model group, the contents of γ-aminobutyric acid in the hypothalamus of rats in the tiaogart, jianpi, buzhen and ginsenoside groups were markedly decreased [（20.92±4.96）, （15.87±2.90）, （13.84±2.63）, （14.94±3.98）, （10.94±3.68） μmoL/g, P 〈 0.05, P 〈 0.05, P 〈 0.01, P 〈 0.01], while the contents in the hypothalamus were obviously raised [（4.12±1.66）, （4.18±1.04）, （6.67±1.29）, （6.11±0.99）, （6.37±3.78） μmoL/g, P 〈 0.05, P 〈 0.01, P 〈 0.01, P 〈 0.01]. ④ The content changes of taurine in the hypothalamus and hippocampus of rats in each group： Compared with model group, the contents of taurine in the hypothalamus of rats in the tiaogart, jiartpi, bushen and ginsenoside groups were markedly decreased [（10.24±1.72）, （7.82±1.14）, （8.00±2.05）, （6.42±3.17） μmoL/g, P 〈 0.05, P 〈 0.05, P 〈 0.01], while the contents in the hippocampus in the jianpi and ginsenoside groups were obviously raised [（12.61±3.51）, （17.03±2.74）, （18.04±2.14） μmoL/g, P 〈 0.01, P 〈 0.01]. CONCLUSION： The central acting site of tioogan recipe may mainly be in the hypothalamus, possibly being related with down-regulating amino acids. While the central acting sites of/iartpi recipe, bushen recipe and ginsenoside may include the hippocampus and hypothalamus, being mainly related with up-regulating amino acids, through enhancing the integration of the hippocampus on stress so as to gain the effect of anti-injury of stress.
出处
《中国临床康复》
CSCD
北大核心
2005年第48期179-181,共3页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金(No.30000217)
广东省自然科学基金(No.000359)~~
