摘要
背景:由于慢性疲劳综合征是慢性病,患者难以坚持长时间服用中药汤剂,因此需要不断积累针对慢性疲劳综合征的专方治疗经验。怡欣乐口服液是在滋阴补肾,填精怡神原则下,选用西洋参、冬虫夏草、龟胶、山萸肉等药物,采用现代制药技术研制而成的中药复方制剂。目的:建立慢性应激性疲劳动物模型,观察怡欣乐口服液对大鼠行为和神经内分泌的影响。设计:随机对照实验。单位:广州军区广州总医院医学实验科,广东省广州市番禺区石碁人民医院妇产科。材料:实验于2003-08/12在广州军区总医院实验中心完成。选取昆明种小鼠60只,随机分为5组,即正常组、模型组、复方阿胶浆组、怡欣乐小剂量组、怡欣乐大剂量组,12只/组。怡欣乐口服液(广州军区总医院医学实验科研制,每1mL相当于生药1g,批号030618)。复方阿胶浆(山东东阿阿胶股份有限公司生产,批号030425)。水迷宫(由中国医学科学院药物研究所提供,型号ZG03008)。方法:全部动物置于室温(20±2)℃清洁级动物室中,5只/笼,自由进食饮水,适应1～2d后开始造模。除正常对照组外,其余4组每天分别在不同时间于(10±1)℃冷水中游泳,每次持续时间为6.0～9.5min,同时复方阿胶浆组给予复方阿胶浆50g/(kg·d),怡欣乐小剂量组给予怡欣乐口服液25g/(kg·d),怡欣乐大剂量组给予怡欣乐口服液50g/(kg·d),正常组及模型组给予生理盐水0.5mL/次,2次/d,共9d。造模完成后,各组进行自发活动(开阔法测定正中格停留时间、穿格次数、竖起或修饰时间、竖起或修饰次数)和力竭游泳时间测定。然后各组小鼠颈动脉取血,麻醉后断头处死,无菌条件下迅速取出下肾上腺组织,收集标本。血清中单胺类神经递质通过高效液相色谱系统和睦电化学检测器检测,肾上腺维生素C含量采用2,4-二硝基苯肼比色法测定。主要观察指标:各组小鼠治疗后自发活动、力竭游泳时间、肾上腺维生素C含量、血清单胺类递质及其代谢产物含量的变化。结果:实验纳入60只小鼠全部进入结果分析。①各组小鼠治疗后正中格停留时间变化:与正常对照组比较,模型组明显延长[(6.64±3.73),(13.80±6.70)s,P<0.01];与模型组比较,复方阿胶浆组、怡欣乐小、大剂量组均显著缩短[(13.80±6.70),(5.12±2.62),(4.51±1.43),(3.34±2.01)s,P<0.05,P<0.05,P<0.01];怡欣乐小、大剂量组明显短于复方阿胶浆组(P<0.05,P<0.01)。②各组小鼠冷水应激后力竭游泳时间比较:与正常对照组比较,模型组明显缩短[(176.44±38.02),(145.01±59.51)min,P<0.01];与模型组比较,复方阿胶浆组、怡欣乐小、大剂量组均显著延长[(145.01±59.51),(172.73±71.59),(181.91±38.60),(186.59±50.81)min,P<0.05,P<0.05,P<0.01];怡欣乐小、大剂量组明显长于复方阿胶浆组(P<0.05,P<0.01)。③各组小鼠冷水应激后肾上腺维生素C含量比较:与正常对照组比较,模型组明显降低[(3951±280),(3546±408)μg/g,P<0.01];与模型组比较,复方阿胶浆组、怡欣乐小、大剂量组均显著延长[(3546±408),(3978±288),(4068±672),(4248±704)μg/g,P<0.05,P<0.05,P<0.01];怡欣乐大剂量组明显长于复方阿胶浆组(P<0.05)。④各组小鼠冷水应激后血清单胺类递质及其代谢产物含量比较:与正常对照组比较,模型组肾上腺素及多巴胺均明显升高[(175±56),(258±97);(1804±889),(4049±1443)nmol/L;P均<0.01],5-羟吲哚乙酸下降[(129.05±34.19),(117.78±42.86)nmol/L,P<0.05];与模型组比较,复方阿胶浆组、怡欣乐小、大剂量组肾上腺素及多巴胺均明显下降[(258±97),(256±135),(230±113),(198±88)nmol/L,P<0.05,P<0.01;(4049±1443),(3702±1266),(2630±939),(1903±658)nmol/L,P<0.05,P<0.01,P<0.01],5-羟吲哚乙酸均明显升高[(117.78±42.86),(138.57±50.43),(155.07±35.31),(236.21±49.82)nmol/L,P<0.05,P<0.05,P<0.01];怡欣乐小、大剂量组肾上腺素及多巴胺均明显低于复方阿胶浆组(P<0.05,P<0.01),且怡欣乐大剂量组5-羟吲哚乙酸显著高于复方阿胶浆组(P<0.01)。结论:怡欣乐口服液可增加疲劳模型小鼠的自发活动、力竭游泳时间及肾上腺素含量,调节血清中单胺类递质系统的失衡,起到抗疲劳、抗应激和调节神经内分泌功能的效应。
BACKGROUND： Chronic fatigue syndrome （CFS） is a chronic disease, with which the patients have difficulty in taking traditional Chinese medicine decoction. So it is necessary to accumulate the special prescriptions for CFS. Based on the principle of nourishing-yin-and-replenishingkidney and filling-up-essence-and-producing-an-effect-toward-clear-vision, American ginseng, aweto, Tortoise Shell and comus officinalis were selected and were made into a compound preparation by using modem pharmaceutical technique. OBJECTIVE： To establish the animal model of chronic stress-related fatigue for observations on the effects of yi xin le oral Liquid （YXL） on behavioral and neuroendocrinological changes in rats. DESIGN： It was a randomized controlled study. SETTING： Medical Study Department, Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA; Department of Obstetrics and Gynecology, Panyu-shiqi People＇s Hospital of Guangzhou city of Guangdong Province MATERIALS： The experiment was conducted at the Laboratory Center of Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA from August to December 2003. Sixty Kunming mice were selected and divided into 5 groups, as normal group, model group, Ejiao （E J） compound group, small dose YXL group and large dose YXL group, with 12 mice in each group. YXL was produced by the Medical Research Department of Guangzhou LiuHuaQiao Hospital （lmL YXL was made from lg raw materials, with a lot number of 030615）. EJ compound was produced by Shandong Dong-E E-Jiao CO.,LTD, with lot number of 030425. Water maze was supplied by Institute of Materia Medica, Chinese Academy of Medical Sciences. METHODS： All the animals were fed in clean level room at （20±2）℃, five mice in one cage, with free food and water supply. After 1-2 days, they were used for establishing animal model. Except for mice in normal group, mice in other groups swam in cold water at （ 10±1 ）℃, in duration of 6.0-9.5 minutes for each time. Additionally, in EJ group, mice received EJ at a dose of 50 g/（kg·d）. In small dose YXL .group, mice received YXL at a dose of 25 g/（kg·d）. In large dose of YXL group, mice received YXL at a dose of 50 g/（kg·d）. In normal group and model group, mice received saline at a dose of 0.5 mL for each time, twice a day and in a duration of 9 days. After establishing the animal model, spontaneous exploratory activity of mice and the forced-swim time was assessed （time spent in the middle of open field or to traversing the field, times to traversing the test field, time spent in orientating while upright or decoration and exploring times before orientating while upright or decoration were measured by open field test）. After tests, blood samples were taken from carotid artery and the animals were decollated under anesthesia. Immediately, their adrenal glands were taken out under axenic condition. And the serum level of monoamine neurotransmitter was measured by high performance liquid chromatography HPLC and electrochemical devices. The concentration of vitamin C in adrenal gland was detected using 2,4 dinitrophenltlhydrazine colorimetric method. MAIN OUTCOME MEASURES： After treatment, the spontaneous exploratory activity, and the forced-swim time, the vitam/n C content in adrenal gland and the changes in monoamine neurotransmitter and itsmetabolites were measured. RESULTS： All the 60 mice in the experiment entered the statistical analysis. ①Comparison of changes in the time spent in the middle of the open field among all the groups： Compared with control group, significantly longer time was spent in model group [（6.64±3.73）, （13.80±6.70） s,P 〈0.01]. Compared with model group, significantly shorter time was spent in EJ group, small dose YXL group and large dose YXL group [（13.80± 6.70）, （5.12±2.62）, （4.51±1.43）, （3.34±2,01）s,P 〈.005,P 〈 0.05,P 〈 0.01]. Compared with EJ group, significantly shorter time was spent in small dose YXL group and large dose YXL grnup（P 〈 0.05 ,P 〈 0.01 ）. ② Comparison of the forced-swim time after cold water stress among all the groups：Compared with control group, significantly shorter time was spent in model group [（176.44±38.02）, （ 145.01±59.51 ）minutes, P 〈 0.01]. Compared with model group, significantly longer time was spent in EJ group, small dose YXL group and large dose YXL group [（145.01±59.51）, （ 172.73 ±71.59）, （ 181,91±38.60）, （ 186.59±50.81 ）minutes,P 〈 0.05 ,P 〈 0.05,P 〈 0.01]. Compared with EJ group, significantly longer time was spent in small dose YXL group and large dose YXL group （P 〈 0.05,P 〈 O.O1 ）.③ Comparison of the vitamin C content in adrenal gland after cold water stress in all the groups： Compared with control group, significantly lower vitamin C content in adrenal gland was detected in model group [（3 951±280）, （3 546±408） μg/g,P 〈 0.01]. Compared with model group, significantly higher vitamin C content in adrenal gland was detected in EJ group, small dose YXL group and large dose YXL group [（3546±408）, （3 978±288）, （4 068±672）, （4 248±704） μg/g,P 〈 0.05,P 〈 0.05,P 〈 0.01]. Compared with EJ group, significantly higher vitamin C content in adrenal gland was detected in large dose YXL group （P 〈 0.05）.④ Comparison of the levels of monoamine neurotransmitter and its metabolites after cold water stress in all the groups： Compared with control group, adrenalin and dopamine were significantly higher in model group [（175±56） nmol/L, （258±97） nmol/L; （1 804±889） nmol/L, （4 049±1 443） nmol/L, P 〈 0.01], while 5-Hydroxyindole Acetic Acid （5-HIAA） was decreased [（129.05±34.19）, （117.78±42.86） nmol/L,P 〈 0.05]. Compared with model group, adrenalin and dopamine were significantly lower in EJ group, small dose YXL group and large dose YXL group [（258±97）, （256±135）, （230±113 ）, （198±88） nmol/L,P 〈 0.05 ,P 〈 0.01 ; （4 049±1443 ）, （3 702 ±1 266）, （2 630±939）, （1 903±658） nmol/l.,P 〈 0.05,P 〈 0.01,P 〈 0.01], while 5-HIAA was significantly higher [（ 117.78±42.86 ）, （138.57± 50.43）, （155.07±35.31）, （236.21±49.82） nmol/L, P 〈 0.05 ,P 〈 0.05 ,P 〈 0.01]. Compared with EJ group, adrenalin and dopamine were significantly lower in small dose YXL group and large dose YXL group （P 〈 0.05 ,P 〈 0.01 ）, and compared with EJ group, the 5-H1AA was significantly higher in large dose YXL group （P〈 0.01）. CONCLUSION： YXL can promote the spontaneous exploratory activity, the forced-swim time and the adrenalin level in mice CFS model. It can also regulate the imbalance of serum monoamine neurotransmitter level and hereby to have the benefits of anti-fatigne, anti-stress and regulations on neuroendocrinological system.
出处
《中国临床康复》
CSCD
北大核心
2005年第36期179-181,共3页
Chinese Journal of Clinical Rehabilitation
