AWA对晕动病的预防作用.doc

AWA对晕动病的预防作用摘要: 目的 通过建立大鼠晕动病模型，考察AWA对晕动病的预防作用 方法 将36只大鼠随机分为正常对照组、晕动病组、阳性药物对照组、低剂量AWA组、中剂量AWA组、高剂量AWA组。以异食量，大鼠行为改变等晕动病指标及肾上腺组织Vc含量变化,判断AWA预防晕动病的效果。 结果 7天内平均异食高岭土量，实验药组与正常组、阳性药物对照组大鼠间无显著差异，且各组均显著低于晕动组。AWA组大鼠肾上腺组织Vc含量明显高于晕动组。 结论 AWA预防晕动病的效果初步得到证实。关键词: 晕动病；异食癖；肾上腺；维生素C The Precaution Effect of AWA on Motion SicknessAbstract: Objective To study the precaution effect of AWA on motion sickness. Methods 36 rats were divided into 6 groups， the normal group, the motion sickness group, the positive drug control group（2mg/ml0.5ml）ratonceday、lower dosage of AWA group （5mg/m0.5m）ratonceday、middle dosage of AWA group （10mg/ml0.5ml）ratonceday、high dosage of AWA 20mg/m0.5ml/ ratonceday. Appropriateness of Kolin consumption as an index of motion sickness in the rat, the exchange of habits and content of adrenaline vitamin C were also taken as the indexes of the precaution effect of AWA on rats curding motion sickness. Results The average amounts of gaoling earth were eaten by each rat in 7days:there were no significant differences amoung the test drug groups, the normal group and the positive group, each group was significantly less than the group of motion sickness. Conclusion The precaution effect of AWA on motion sickness was proved preliminarily.Key words: motion sickness; appropriateness; adrenaline; vitamin C 晕动病是人们日常生活中常见的一种症候群，虽已有百余年的研究历史，但迄今对晕动病的预防还未取得重大突破1。目前，市场上的预防药物多为镇静催眠药和抗组织胺类药物，副作用大，锥体外系作用较强，且禁忌症较多。生物波研究中心在进行药物试验过程中偶然发现，促波氨基糖（accelerating wave amino sugar, AWA）有预防晕动病的作用，特别是对晕车、船者反映出前所未有的控制效果，同时未发现嗜睡，不适等副作用。为明确其预防晕动病的效果及建立效果评定的指标，选用SD大鼠建立晕动病模型，以异食高岭土量的变化来判定症状产生及程度2，同时以行为、体征改变及肾上腺组织维生素C（vitamin C, Vc）含量变化为实验指标，来证实AWA的预防效果。材料与方法1 材料1.1实验动物 成年SD大鼠36只（第三军医大学动物所），体重170220g，雌雄各半，常规饲养。1.2实验材料高岭土（苏州中国高岭土公司） 具体配方：1kg高岭土粉、10g阿拉伯树胶，加水搅拌形成食物形状类似物，于80烘干后，与饲料同时放入饲养笼内。 Vc测试试剂盒、考马斯亮兰及标准蛋白（南京建成生物工程研究所）。盐酸地芬尼多（湖南千金湘江药业060107）。晕动模拟器（自制、全程（0250V）调压器、小方桶（30cm30cm20cm）。其余试剂均为市售，常规配制。2方法2.1大鼠模拟晕动刺激方法2.1.1将30只SD大鼠随机分为正常对照组，晕动病模型组，低剂量实验组（5mg/ml0.5ml），中剂量实验组（10mg/ml0.5ml），高剂量实验组（20mg/ml0.5ml），阳性药物对照组，每组6只。2.1.2刺激方法：给药组大鼠每天灌胃一次，给药30分钟后，将大鼠无束缚放到模拟器中，以180r/min恒速持续刺激15min，每天一次。2.2指标测定方法2.2.1大鼠接受模拟刺激后的行为变化观测，采用每天定时观测的方法。2.2.2大鼠异食高岭土量测定：大鼠没有呕吐反射，但在受到异常运动刺激后，会出现一种类似于呕吐反射的行为，即异食癖3。因此大鼠异食高岭土的量，一定程度上反映了晕动病症状是否产生及症状程度。实验开始后，于每天8：00记录24小时内进食高岭土量。2.2.3大鼠肾上腺组织Vc及蛋白质含量的测定连续刺激6日后，分批处死各鼠，取肾上腺，称重，加入9倍重量的生理盐水匀浆，取匀浆液离心，2000rpm,3min，取上清液进行Vc含量测定，具体操作按试剂盒说明进行。计算公式：Vc(g/ml)=（样品管OD-空白管OD）/（标准管OD-空白管OD）64。蛋白质含量（P）测定 另取上清适量，稀释10倍，取0.5ml，用考马斯亮兰显色，测OD值，以标准蛋白（0.615mg/ml）为对照，=595nm，计算蛋白含量。P(mg/ml)=（样品管OD-空白管OD）/（标准管OD-空白管OD）0.61510。结果表示：Vc(g)/ P(mg)。结 果1大鼠接受模拟刺激后的行为变化：模拟刺激结束放回笼后，晕动组大鼠表现出反应迟钝，活动量明显减少，无进食饮水行为，全身毛皮蓬松，排尿、排便次数增加等中枢抑制反应。给AWA的各实验组动物及给盐酸地芬尼多预防组动物状态较好，排尿、排便次数无明显改变。2各实验组大鼠进食高岭土量变化情况见表1。表1 晕动实验中各组大鼠进食高岭土情况组 别刺激后不同时间（d）大鼠进食高岭土量（g）123456正常组 0.1370.2080.4150.2750.480.023晕动组 0.0370.3370.8220.4670.9370.698低剂量AWA组0.1820.1750.2370.1350.2120.247中剂量AWA组0.3000.1780.2280.2240.3970.366高剂量AWA组0.1380.1000.0010.0850.1680.390阳性药物对照组0.200.310.280.330.320.40从表中可以看出，AWA很好的控制了晕动症状的发生，且控制效果与剂量呈较大正相关性。结果用线形图表示见图1。图1 晕动实验中各组大鼠进食高岭土量的线形图从图中可以看出，晕动组进食高岭土量远大于正常组及给药各组（p0.05），且均在正常值范围内（0.150.3g）/天。3各实验组大鼠肾上腺组织的Vc含量测定结果，见表2、图2。表2 晕动实验中各组大鼠Vc含量组 别大鼠肾上腺组织Vc含量（g）正常组3.850.163晕动组3.110.162阳性药物组3.40.266AWA组均值 3.530.183图2 各组大鼠肾上腺组织Vc含量统计直方图由统计结果可以看出： AWA控制组Vc含量明显高于晕动组（p0.05）,比正常组略低（p0.05）。讨 论大鼠没有呕吐反应，但在接受晕动刺激后可出现类似于呕吐反应的异食癖行为。本研究从这一点出发，以异食行为的改变作为大鼠晕动病发生及其程度的指标之一。另外本文就各实验组大鼠肾上腺组织的Vc含量进行了交叉性比较。针对晕车、船发生的内分泌异常机制，涉及到晕动病发生时激素和神经递质的水平升高6，7。经常用药物治疗，机体神经-内分泌系统趋于正常。本实验采用AWA对实验动物模型进行预防模拟晕动刺激实验，结果表明，给AWA药物的大鼠其晕动病症状虽没有完全消失,但程度(相关检测指标)比晕动组明显减轻,也比阳性药物对照组症状轻，AWA实验组大鼠肾上腺组织中Vc含量比正常鼠偏低,显著高于阳性药物对照组。肾上腺组织Vc含量作为药物预防晕动病效果指标，表明试验药物AWA对晕动病有确切预防效果，而且在一定剂量范围内效果显出量效关系。AWA作为治疗其它疾病药物的毒理学实验已证明无明显毒副作用。因此，AWA有可能成为晕动病预防中的安全而特效的药物。 参考文献:1 Lang IM,Sarna SK,Shaker R. Gastrointestinal motor and myoelectric correlates of motion sickness. Am J Physiol,1999,277(3Pt1):G642-652.2 吉雁鸿,郭俊生,李敏,等.大鼠晕船适应动物模型的建立.中华劳动卫生职业病杂志 2004年12月第22卷第6期.3 Uno A,Takeda N,Horili A,et al.Histamine release from the hypothalamus musinduced by gravity change in rats and space motion sickness. Physiol Behav ,1997,61:883-8874 Kohl RL, Endocrine correlates of susceptibility to motion sicknessJ. Aviat space Environ Med,1985,56(12):1158-1165.5 McCaffrey RJ.Appropriateness of Kolin consumption as an index of motion sickness in the rat.Physiol Behav,1985,35:151-156.6 Horii A ,Koike ,Uno A,et al.Vestibular modulation of plasma vasopressin levels in rats.Brain Res,2001,914(1-2):179-184.7 Javid FA,Naylor RJ.Opioid receptor involvement in the adaptation to motion sickness in suncus murinus.Pharmacol Biochem Behav,2001,68:761-767.Preventive Effect of AWA on Motion SicknessJunkang Liu* Jie Chen* Hongling Dan* Qiwang Xu* Biowave Research Center, the Third Military Medical University, Chongqing 400038, ChinaAbstract: Objective To study the preventive effect of AWA on motion sickness. Methods 36 rats were divided into 6 groups，the normal group, the motion sickness group, the positive drug control group, lower dosage of AWA group, middle dosage of AWA group and high dosage of AWA. Heterophile consumption of Kaolin as an index of motion sickness in the rat, the change of behavior and content of vitamin C in adrenal were also taken as the indexes of the preventive effect of AWA on rats motion sickness. Results The average amounts of Kaolin were eaten by each rat in 7days: there were no significant differences among the test drug groups, the normal group and the positive group, each group was significantly less than the group of motion sickness. Conclusion The preventive effect of AWA on motion sickness was proved preliminarily.Key words: Motion sickness; Heterophile consumption; Adrenal; Vitamin C The motion sickness is a common disease of hundred-year history of research, but up to now there is no important breakthrough in the treatment of the disease 1, 2. The drugs sold in market recently to prevent the motion sickness are mainly the drugs of sedation and hypnogenesis and the anticholinergic agents, which have comparatively great side effects, strong effect on extrapyramidal system and many contraindications. During the new drug test, It was found occasionally that the accelerating wave amino-sugar (AWA) exhibits the effects of preventing motion sickness, esp. the unmatched prevention effects on the carsickness, seasickness, Meantime, without the side-effects of lethargy, maladjustment. For clarifying the effects of preventing motion sickness, the indexes were selected to evolutes the effects. This paper tries to prove the preventive effectiveness of AWA by establishing model with the selected SD rats, by judging the creation and degree of symptom through the variation of the amount of Kaolin eaten by the rats and by taking the variations of behavior and sigh and the variations of some neuro-endocrine factors as the objective indexes 3, 4.Materials and methods1. Animals36 health SD rats are selected, of which half male and half female, with the individual weight ranging from 200 to 250 g. All the rats are purchased from the Animal Research Institute of third Military Medical University. The female rats are not pregnant. Feed all the selected rats in a normal and regular way.2. MaterialsKaolin (Suzhou Kaolin Company): Mixing of 1 kg of Kaolin with 10g Arabic gum to produce an object similar in shape to food after adding water and stirring. Dry it at the temperature of 80 and put it together with fodder in rat cages. Vitamin C test reagent kit, produced by Nanjing Jiancheng Bioengineering Research Institute. Hydrochloric acid difenidol, produced by Hunan Qianjin Xiangjiang Pharmaceutical Industry, batch number: 060107. Motion sickness simulator, self-made, full-range: 0250V, pressure regulator and small barrel (30cm30cm20cm). All other reagents are commercial ones and are prepared conventionally. Corresponding Author Present Address: Biotimes Laboratory, Beijing 101200, ChinaE-mail: 3. Stimulation method of motion sickness simulation of rats3.1 Divide SD rats (half male and half female) randomly into the following groups, with 6 rats in each individual group: normal group, motion sickness remarkable group, lower dosage control group (5mg/ml 0.5ml), middle dosage control group (10mg/ml0.5ml), higher dosage control group (20mg/ml0.5ml) and positive drug control group.3.2 Stimulation method: Perform stomach perfusion to the rats of medicine taken group. 30minutes after taking medicine, put rats that are free to move in a simulator and perform the successive stimulation at the constant speed of 180r/min for 15 minutes, once a day.4. Index determination method4.1 Adopt the periodic observation method to observe the habit changes of rats after receiving the standard stimulation everyday.4.2 Determination of amount of Kaolin eaten by rats: Rats do not have vomit reflex, but they will have a reflex behavior similar to vomit, called heterophile feature after experiencing abnormal stimulation of motion. Therefore, the amount of Kaolin eaten by rats can reflect at a certain degree the emerging of a symptom as well as the degree of a symptom 5. Take down notes about the amount of Kaolin eaten in 24 hours at 8:00 everyday after the experiment is started.5. Determination of adrenaline vitamin C of rats Kill the experimented rats in groups after six-day successive stimulation. Take the adrenal and perform homogenization as per (tissue: normal saline = 1:9). Centrifugalize it at 2000rpm3 min, take supernatant fluid to determine the content of Vitamin C. The procedure can be conducted according to the instruction of the Assay kit. Calculate the content of Vitamin C and express the Vitamin C content in the following formulas: Vit C = (sample tube OD blank tube OD) / (standard tube OD blank tube OD)64; Results1. Behavior change of rats after receiving simulated stimulation After they were put back into the animal room at the end of simulation, the rats in the non-medicine taking group showed that they were slow in reaction, the amount of motion decreases remarkable, without eating and drinking, the fur throughout the whole body became fluffy and the central inhibition reaction including the time increase of urination and defecation was remarkable. However, It was observed that the rats in the preventive group of taking AWA experiment drug and hydrochloric acid difenidol were in a comparatively good state and the times of urination and defecation exhibits no significant variation.2. The changes of rats in consumption of KaolinTable 1 Eating of kaolin by rats in individual group in motion sickness experimentGroupsThe amount (g) of Kaolin eaten by the rats at different time after stimulation123456Normal group0.1370.2080.4150.2750.480.023Motion sickness group0.0370.3370.8220.4670.9370.698Low dosage AWA group0.1820.1750.2370.1350.2120.247Middle dosage AWA group0.3000.1780.2280.2240.3970.366high dosage AWA group0.1380.1000.0010.0850.1680.390Positive drug group0.200.310.280.330.320.40From the Table1, it can be seen that AWA controls in a very good way the occurrence of motion sickness and the control effect has a great positive relation with the dosage taken. The result is expressed in a line diagram. See the following figure 1.Figure 1 String diagram of Kaolin eaten by rats in individual group in motion sickness experimentIt can be seen remarkably through the diagram that the amount of eaten by the rats in motion sickness group is largely greater than the amount taken by the rats in normal group and the rats in each AWA-taken group (p0.05) and they are all within the range of normal values (0.150.3g)/ day times.3. The determination of vitamin C content in adrenal of rats The results show in Table 2 and Figure 2Table 2: Vitamin C content of rats in individual group in motion sickness experiment GroupsValue rangeNormal Group3.850.163Motion Sickness Group3.110.162Positive Drug Group3.40.266Experiment Drug Group3.530.183Figure 2: Block diagram of adrenal vitamin C content of rats in individual group It can be seen through the statistic results that the Vitamin C content of the rats in experiment drug (AWA) control group is remarkably higher than that of the rats in motion sickness group (p0.05) and is a little bit higher than the Vitamine C content of the rats in positive drug group, but a little bit lower than the Vitamine C content of the rats in normal group (p0.05).DiscussionRats do not have vomit reflex, but they will have a reflex behavior similar to vomit, called heterophile feature after experiencing the motion sickness stimulation. The experiment mentioned in this paper starts from this point of view, and takes the change of the heterophile feature as one of the objective indexes showing the occurrence and degree of motion sickness of rats. In addition, a thorough comparison of the adrenal Vitamin C content of rats in each individual experiment group has been made in this paper. Considering that the mechanism of motion sickness involves the abnormal of endocrine function, with the higher level of hormone and neuro-transmitters 6-7, which is on the basis of the cryptorrhea presumption of the nerve non-matching of the theory of motion sickness occurrence mechanism, i.e. the level of many hormones and neurotransmitters at occurrence of motion sickness rises up extraordinarily and the in vivo neuroendocrine system of the organism will have a series of changes during drug treatment. The group division in the experiment is reasonable, with a clear positive comparison drug and negative comparison. Further, the experiment drug is taken in different dosages. It has been proven through experiment that the symptom of motion sickness of the rats having taken AWA drug has not gone away completely, however, it has been alleviated remarkably as compared with the rats in the motion sickness group. It is also the case if we compare the result with the symptom of motion sickness of the rats in the experiment group taking the medicine of dizziness termination. The adrenal Vitamin C content of the rats in experiment group is lower than that of the rats in normal group, but is remarkably higher than that of the rats in motion sickness group and positive drug comparison group. Being as an objective index showing the preventive effect of AWA on motion sickness. Moreover, within certain dosage limits, there were the correlations of dose-effects. It had been proved that AWA didnt have obvious toxic action through the toxic experiment when used as the drugs treating other illness and will most likely become a drug of high efficiency but low poison in the treatment of motion sickness.References1. Lang IM, Sama SK, Shaker R. Gastrointestinal motor myoelectric correlates of motion sickness. Am J Physiol, 1999, 277(3Pt1): C624-652.2. Yanhong Ji, Junsheng Guo, Min Li, et al. Animal model establishment of seasickness adaptability of rats. China Journal of Labor Health and Occupational Disease, 2004, 22 (6): 463-464.3. Uno A, Takeda N, Horili A, et al. Histamine release from the hypothalamus musinduced by gravity change in rats and space motion sickness. Physio Behav, 1997, 61: 883-887.4. Kohl RL. Endocrine correlates of susceptibility to motion sickness. Aviat space Environ Med, 1985, 56: 1156-1163.5. McCaffrey RJ. Appropriateness of Kolin consumption as an index of motion sickness in the rat. Physiol Behav, 1985, 35: 151-156.6. Horii A, Koike, Uno A, et al. Vestibular modulation of plasma vasopressin levels in rats. Brain Res, 2001, 914(1-2): 179-184.7. Javid FA, Naylor RJ. Opioid receptor involvement in the adaptation to motion sickness in suncus murinus. Pharmacol Biochem Behav, 2001, 68: 761-767.AWA 对晕动病的预防作用刘俊康* 陈杰* 淡洪林* 徐启旺* 中国人民解放军第三军医大学生物波研究室 4 0 0 0 3 8摘要：目的 通过建立大鼠晕动病模型，考察AWA 对晕动病的预防作用。方法 将36 只大鼠随机分为正常对照组、晕动病组、阳性药物对照组、低剂量AWA 组、中剂量AWA 组、高剂量AWA 组。以异食量、大鼠行为改变等晕动病指标及肾上腺组织VC 含量变化，判断AWA 预防晕动病的效果。结果 7 天内大鼠的平均异食高岭土量，实验药组及正常组与阳性药物对照组相比，无显著差异，且三个组均显著低于晕动组。AWA 组大鼠肾上腺组织VC 含量明显高于晕动组。结论 AWA 预防晕动病的效果初步得到证实。关键词：晕动病；异食癖；肾上腺；维生素C晕动病是人们日常生活中常见的一种症候群，虽已有百余年的研究历史，但迄今对晕动病的预防还未取得重大突破1。目前，市场上的预防药物多为镇静催眠药和抗组织胺类药物，副作用大，锥体外系作用较强，且禁忌症较多。生物波研究中心在进行药物试验过程中偶然发现，促波氨基糖（accelerating wave aminosugar，AWA）有预防晕动病的作用，特别是对晕车、晕船者反映出前所未有的控制效果，同时未发现嗜睡、不适等副作用。为明确其预防晕动病的效果，选用SD大鼠建立晕动病模型，以确定效果评定的指标异食高岭土量的变化来判定症状产生及程度2，以行为、体征改变及肾上腺组织维生素C（vitamin C，VC）的含量变化为实验指标，来证实AWA的预防效果3,4。材料与方法1. 动物成年SD 大鼠36 只，体重170220g，雌雄各半，第三军医大学动物所提供。常规饲养。2. 实验材料高岭土（苏州中国高岭土公司） 具体配方：1kg 高岭土粉、10g阿拉伯树胶，加水搅拌成食物形状类似物，80烘干后，与饲料同时放入饲养笼内。VC测试试剂盒，购自南京建成生物工程研究所。盐酸地芬尼多，批号060107，湖南千金湘江药业生产。晕动模拟器，自制、全程（0250V）调压器、小方桶（30cm 0cm 0cm）。其余试剂均为市售，常规配制。3. 大鼠模拟晕动刺激方法3.1 将36 只SD 大鼠随机分为正常对照组，晕动病模型组，低剂量实验组（5mg/ml 0.5ml），中剂量实验组（10mg/ml 0.5ml），高剂量实验组（20mg/ml 0.5ml），阳性药物对照组，每组6 只。3.2 刺激方法：给药组大鼠每天灌胃1 次，给药30min 后，将大鼠无束缚放到模拟器中，以180r/min 恒速持续刺激15min，每天1 次。4. 指标测定方法4.1 大鼠接受模拟刺激后的行为变化观测：采用每天定时观测、记录的方法。4.2 大鼠异食高岭土量的测定：实验开始后，于每天8:00 记录2