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1、AWA對暈動病的預防作用摘要: 目的 通過建立大鼠暈動病模型,考察AWA對暈動病的預防作用 方法 將36只大鼠隨機分為正常對照組、暈動病組、陽性藥物對照組、低劑量AWA組、中劑量AWA組、高劑量AWA組。以異食量,大鼠行為改變等暈動病指標及腎上腺組織Vc含量變化,判斷AWA預防暈動病的效果。 結果 7天內平均異食高嶺土量,實驗藥組與正常組、陽性藥物對照組大鼠間無顯著差異,且各組均顯著低于暈動組。AWA組大鼠腎上腺組織Vc含量明顯高于暈動組。 結論 AWA預防暈動病的效果初步得到證實。關鍵詞: 暈動?。划愂绸保荒I上腺;維生素C The Precaution Effect of AWA on Mo

2、tion 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/ml×0.5ml)rat·once·day、lower dosage of AWA group (5mg/m×0.5m)rat&

3、#183;once·day、middle dosage of AWA group (10mg/ml×0.5ml)rat·once·day、high dosage of AWA 20mg/m×0.5ml/ rat·once·day. 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

4、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

5、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 暈動病是人們日常生活中常見的一種癥候群,雖已有百余年的研究歷史,但迄今對暈動病的預防還未取得重大突破1。目前,市場上的預防藥物多為鎮(zhèn)靜催眠藥和抗組織胺類藥物,副作用大,錐體外系作用較強,且禁忌癥較多。生物波研究中心在進

6、行藥物試驗過程中偶然發(fā)現(xiàn),促波氨基糖(accelerating wave amino sugar, AWA)有預防暈動病的作用,特別是對暈車、船者反映出前所未有的控制效果,同時未發(fā)現(xiàn)嗜睡,不適等副作用。為明確其預防暈動病的效果及建立效果評定的指標,選用SD大鼠建立暈動病模型,以異食高嶺土量的變化來判定癥狀產生及程度2,同時以行為、體征改變及腎上腺組織維生素C(vitamin C, Vc)含量變化為實驗指標,來證實AWA的預防效果。材料與方法1 材料1.1實驗動物 成年SD大鼠36只(第三軍醫(yī)大學動物所),體重170220g,雌雄各半,常規(guī)飼養(yǎng)。1.2實驗材料高嶺土(蘇州中國高嶺土公司) 具體配

7、方:1kg高嶺土粉、10g阿拉伯樹膠,加水攪拌形成食物形狀類似物,于80烘干后,與飼料同時放入飼養(yǎng)籠內。 Vc測試試劑盒、考馬斯亮蘭及標準蛋白(南京建成生物工程研究所)。鹽酸地芬尼多(湖南千金湘江藥業(yè)060107)。暈動模擬器(自制、全程(0250V)調壓器、小方桶(30cm×30cm×20cm)。其余試劑均為市售,常規(guī)配制。2方法2.1大鼠模擬暈動刺激方法2.1.1將30只SD大鼠隨機分為正常對照組,暈動病模型組,低劑量實驗組(5mg/ml×0.5ml),中劑量實驗組(10mg/ml×0.5ml),高劑量實驗組(20mg/ml×0.5ml),

8、陽性藥物對照組,每組6只。2.1.2刺激方法:給藥組大鼠每天灌胃一次,給藥30分鐘后,將大鼠無束縛放到模擬器中,以180r/min恒速持續(xù)刺激15min,每天一次。2.2指標測定方法2.2.1大鼠接受模擬刺激后的行為變化觀測,采用每天定時觀測的方法。2.2.2大鼠異食高嶺土量測定:大鼠沒有嘔吐反射,但在受到異常運動刺激后,會出現(xiàn)一種類似于嘔吐反射的行為,即異食癖3。因此大鼠異食高嶺土的量,一定程度上反映了暈動病癥狀是否產生及癥狀程度。實驗開始后,于每天8:00記錄24小時內進食高嶺土量。2.2.3大鼠腎上腺組織Vc及蛋白質含量的測定連續(xù)刺激6日后,分批處死各鼠,取腎上腺,稱重,加入9倍重量的生

9、理鹽水勻漿,取勻漿液離心,2000rpm,3min,取上清液進行Vc含量測定,具體操作按試劑盒說明進行。計算公式:Vc(g/ml)=(樣品管OD-空白管OD)/(標準管OD-空白管OD)×6×4。蛋白質含量(P)測定 另取上清適量,稀釋10倍,取0.5ml,用考馬斯亮蘭顯色,測OD值,以標準蛋白(0.615mg/ml)為對照,=595nm,計算蛋白含量。P(mg/ml)=(樣品管OD-空白管OD)/(標準管OD-空白管OD)×0.615×10。結果表示:Vc(g)/ P(mg)。結 果1大鼠接受模擬刺激后的行為變化:模擬刺激結束放回籠后,暈動組大鼠表現(xiàn)出

10、反應遲鈍,活動量明顯減少,無進食飲水行為,全身毛皮蓬松,排尿、排便次數增加等中樞抑制反應。給AWA的各實驗組動物及給鹽酸地芬尼多預防組動物狀態(tài)較好,排尿、排便次數無明顯改變。2各實驗組大鼠進食高嶺土量變化情況見表1。表1 暈動實驗中各組大鼠進食高嶺土情況組 別刺激后不同時間(d)大鼠進食高嶺土量(g)123456正常組 0.1370.2080.4150.2750.480.023暈動組 0.0370.3370.8220.4670.9370.698低劑量AWA組0.1820.1750.2370.1350.2120.247中劑量AWA組0.3000.1780.2280.2240.3970.366高劑

11、量AWA組0.1380.1000.0010.0850.1680.390陽性藥物對照組0.200.310.280.330.320.40從表中可以看出,AWA很好的控制了暈動癥狀的發(fā)生,且控制效果與劑量呈較大正相關性。結果用線形圖表示見圖1。圖1 暈動實驗中各組大鼠進食高嶺土量的線形圖從圖中可以看出,暈動組進食高嶺土量遠大于正常組及給藥各組(p<0.05);其它各組與正常組無統(tǒng)計學差異(P>0.05),且均在正常值范圍內(0.150.3g)/天。3各實驗組大鼠腎上腺組織的Vc含量測定結果,見表2、圖2。表2 暈動實驗中各組大鼠Vc含量組 別大鼠腎上腺組織Vc含量(g)正常組3.85&#

12、177;0.163暈動組3.11±0.162陽性藥物組3.4±0.266AWA組均值 3.53±0.183圖2 各組大鼠腎上腺組織Vc含量統(tǒng)計直方圖由統(tǒng)計結果可以看出: AWA控制組Vc含量明顯高于暈動組(p<0.05),略高于陽性藥物對照組,但無統(tǒng)計學差異(P>0.05),比正常組略低(p<0.05)。討 論大鼠沒有嘔吐反應,但在接受暈動刺激后可出現(xiàn)類似于嘔吐反應的異食癖行為。本研究從這一點出發(fā),以異食行為的改變作為大鼠暈動病發(fā)生及其程度的指標之一。另外本文就各實驗組大鼠腎上腺組織的Vc含量進行了交叉性比較。針對暈車、船發(fā)生的內分泌異常機制,涉

13、及到暈動病發(fā)生時激素和神經遞質的水平升高6,7。經常用藥物治療,機體神經-內分泌系統(tǒng)趨于正常。本實驗采用AWA對實驗動物模型進行預防模擬暈動刺激實驗,結果表明,給AWA藥物的大鼠其暈動病癥狀雖沒有完全消失,但程度(相關檢測指標)比暈動組明顯減輕,也比陽性藥物對照組癥狀輕,AWA實驗組大鼠腎上腺組織中Vc含量比正常鼠偏低,顯著高于陽性藥物對照組。腎上腺組織Vc含量作為藥物預防暈動病效果指標,表明試驗藥物AWA對暈動病有確切預防效果,而且在一定劑量范圍內效果顯出量效關系。AWA作為治療其它疾病藥物的毒理學實驗已證明無明顯毒副作用。因此,AWA有可能成為暈動病預防中的安全而特效的藥物。 參考文獻:1

14、 Lang IM,Sarna SK,Shaker R. Gastrointestinal motor and myoelectric correlates of motion sickness. Am J Physiol,1999,277(3Pt1):G642-652.2 吉雁鴻,郭俊生,李敏,等.大鼠暈船適應動物模型的建立.中華勞動衛(wèi)生職業(yè)病雜志 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

15、 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

16、 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 S

17、icknessJunkang 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

18、 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 prevent

19、ive 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

20、 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 dise

21、ase 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 wa

22、s 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

23、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

24、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 th

25、e 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 addi

26、ng 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: 0

27、60107. Motion sickness simulator, self-made, full-range: 0250V, pressure regulator and small barrel (30cm×30cm×20cm). All other reagents are commercial ones and are prepared conventionally. Corresponding Author Present Address: Biotimes Laboratory, Beijing 101200, ChinaE-mail: biowave20003

28、. 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 gr

29、oup (10mg/ml×0.5ml), higher dosage control group (20mg/ml×0.5ml) 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

30、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

31、 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

32、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 =

33、1:9). Centrifugalize it at 2000rpm×3 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 t

34、ube OD blank tube OD) / (standard tube OD blank tube OD)×6×4; 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, th

35、e 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 A

36、WA 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

37、 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 group0.1

38、380.1000.0010.0850.1680.390Positive 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

39、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

40、 rats in each AWA-taken group (p<0.05). There are no statistic difference if we compare the result of the rest groups with that of the normal group (p>0.05) 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 r

41、ats The results show in Table 2 and Figure 2Table 2: Vitamin C content of rats in individual group in motion sickness experiment GroupsValue rangeNormal Group3.85±0.163Motion Sickness Group3.11±0.162Positive Drug Group3.4±0.266Experiment Drug Group3.53±0.183Figure 2: Block diagra

42、m 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 (p<0.05) and is a little bit higher than the Vi

43、tamine 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 (p<0.05). However, there is no difference in the statistic point of view (p>0.05).DiscussionRats do not have vomit reflex, but they will have a reflex behavior si

44、milar 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 o

45、f 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

46、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 orga

47、nism 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 si

48、ckness 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 me

49、dicine 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 preventiv

50、e 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 e

51、fficiency 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 adapta

52、bility 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

53、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

54、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 對暈動病的預防作用劉俊康* 陳杰* 淡洪林* 徐啟旺* 中國人民解放軍第三軍醫(yī)大學生物波研究室 4 0 0 0 3 8摘要:目的 通過建立大鼠暈動病模型,考察AWA 對暈動病的預防作用。方法 將36 只大鼠隨

55、機分為正常對照組、暈動病組、陽性藥物對照組、低劑量AWA 組、中劑量AWA 組、高劑量AWA 組。以異食量、大鼠行為改變等暈動病指標及腎上腺組織VC 含量變化,判斷AWA 預防暈動病的效果。結果 7 天內大鼠的平均異食高嶺土量,實驗藥組及正常組與陽性藥物對照組相比,無顯著差異,且三個組均顯著低于暈動組。AWA 組大鼠腎上腺組織VC 含量明顯高于暈動組。結論 AWA 預防暈動病的效果初步得到證實。關鍵詞:暈動??;異食癖;腎上腺;維生素C暈動病是人們日常生活中常見的一種癥候群,雖已有百余年的研究歷史,但迄今對暈動病的預防還未取得重大突破1。目前,市場上的預防藥物多為鎮(zhèn)靜催眠藥和抗組織胺類藥物,副作

56、用大,錐體外系作用較強,且禁忌癥較多。生物波研究中心在進行藥物試驗過程中偶然發(fā)現(xiàn),促波氨基糖(accelerating wave aminosugar,AWA)有預防暈動病的作用,特別是對暈車、暈船者反映出前所未有的控制效果,同時未發(fā)現(xiàn)嗜睡、不適等副作用。為明確其預防暈動病的效果,選用SD大鼠建立暈動病模型,以確定效果評定的指標異食高嶺土量的變化來判定癥狀產生及程度2,以行為、體征改變及腎上腺組織維生素C(vitamin C,VC)的含量變化為實驗指標,來證實AWA的預防效果3,4。材料與方法1. 動物成年SD 大鼠36 只,體重170220g,雌雄各半,第三軍醫(yī)大學動物所提供。常規(guī)飼養(yǎng)。2.

57、 實驗材料高嶺土(蘇州中國高嶺土公司) 具體配方:1kg 高嶺土粉、10g阿拉伯樹膠,加水攪拌成食物形狀類似物,80烘干后,與飼料同時放入飼養(yǎng)籠內。VC測試試劑盒,購自南京建成生物工程研究所。鹽酸地芬尼多,批號060107,湖南千金湘江藥業(yè)生產。暈動模擬器,自制、全程(0250V)調壓器、小方桶(30cm× 0cm × 0cm)。其余試劑均為市售,常規(guī)配制。3. 大鼠模擬暈動刺激方法3.1 將36 只SD 大鼠隨機分為正常對照組,暈動病模型組,低劑量實驗組(5mg/ml × 0.5ml),中劑量實驗組(10mg/ml ×0.5ml),高劑量實驗組(20mg/ml × 0.5ml),陽性藥物對照組,每組6 只。3.2 刺激方法:給藥組大鼠每天灌胃1 次,給藥30min 后,將大鼠無束縛放到模擬器中,以180r/min 恒速持續(xù)刺激15min,每天1 次。4. 指標測定方法4.1 大鼠接受模擬刺激后的行為變化觀測:采用每天定時觀測、記錄的方法。4.2 大鼠異食高嶺土量的測定:實驗開始后,于每天8:00 記錄24h 內大鼠進食高嶺土量。5. 大鼠腎上腺組織

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