基于浓氨水引咳法建立小鼠咳嗽敏感性剔除标准的方法学研究_杨利娟

,:1.。2.。3.,。:1.、。2.,。　　Ⅰ、Ⅱ、,,,。1　DosenkoVE,NagibinVS,TumanovskayaLV,etal.Postconditioningpre-ventsapoptoticnecroticandautophagiccardiomyocytecelldeathinculture.Fi-ziolZh,2005;51(3)∶12～172　RohrS,SchollyDM,KleberAG,etal.Patternedgrowthofneonatalratheartcellsinculture.Morphologicalandelectrophysiologicalcharacterization.CircRes,1991;68(1)∶114～1303　WeisenseeD,SeegerT,BittnerA,etal.Coculturesoffetalandadultcar-diomyocytesyieldrhythmicallybeatingrodshapedheartcellsfromadultrats.InVitroCellDevBiolAnim,1995;31(3)∶190～1954　KonoT,PollmanM,CorvolP,etal.Roleofcollagenaseandotherprote-olyticenzymesinthedispersalofanimaltissues.BiochimBiophysActa,1968;178∶397～4005　LiC,CaoL,ZhengQ,etal.Taurinemaypreventdiabeticratsfromdevel-opingcardiomyopathyalsobydownregulatingangiotensinIItype2receptorex-pression.CardiovascDrugsTher,2005;19(2)∶105～1206　,,,.52BrdU.,2008;20(5)∶566～5697　MitchesonJS,HancoxJC,LeviAJ,etal.Culturedadultcardiacmyco-cytes:futureapplications,culturemethods,morphologicalandelectrophysio-logicalproperties.CardiovascRes,1998;39(2)∶280～300AnmethodforprimarycultureofneonateratcardiacmyocytesZhangLina,JinGuoqin(BiochemistryDepartmentinShanghaiUniversityofTraditionalChineseMedicine,Shanghai　201203)　　Objective∶Toexploretheconditionsandapproachtoculturepurifiedneonatalratcardiocytes.Methods∶Thetypsinase,collagenaseⅠandcollagenaseⅡwasusedtodigestandisolatemyocardialtissue,andthendifferentialadhesionandchemicalinhibitionwaswasthewayforpurification.Results∶24hourslater,cellsbegantoadhereandSpindle-shapedcellismain.48hourslater,cellsstretchoutthepseudopo-diawithStrongrefractiion;3～4dayslater,Pseudopodiaareincontacttoanetwork,cellclustersformed.60%～80%beatingcardiomyo-cyteswereobservedandthebeatingfrequencyofcellswas50beats/minafter3days.Conclusion∶Thismethodisaneffectivewaytoculti-vatepurifiedneonatalratcardiocyteswithlongsurvivaltime.Keywords∶myocardiocyte;primaryculture;neonaterat杨利娟,王　张,孟宪丽＊,孙苏逖,朱丽娜,李冬琪(成都中医药大学,成都　611137)　　　　:制定小鼠对浓氨水致咳嗽的敏感性剔除标准。:观察13%浓氨水喷雾60s后致小鼠咳嗽的三分钟内咳嗽次数和潜伏期,并根据相应的频数图拟定剔除标准,比较两指标的数据在剔除前后的差异性。:执行剔除标准后,小鼠的咳嗽反应明显增加且三分钟内咳嗽次数的离散度减少,从统计学和实验动物学专业的角度分析,经数据剔除后动物对氨水的刺激更加敏感,咳嗽次数适中,数据集中,有利于药物的有效性评价。:建立并证实了基于浓氨水引咳法的小鼠咳嗽敏感性剔除标准,即三分钟内咳嗽次数少于10次及多于80次者;喷雾后潜伏期小于15秒及大于50秒者;动物死亡、状态不佳、反应异常剧烈者等;符合三者其中任何一项者,即为剔除对象。　　　;;;;　　。,,,,。1　1.1　　,,18～20g,898,,SCXK()2004-15。1502010;26(5)＊通讯作者DOI:10.13412/j.cnki.zyyl.2010.05.0771.2　　(),25%～28%,,2009814。,13%。1.3　　980-A()。1.4　　,50ml(13%)[1,2]。1000ml,,(,)。(MAX)60s,,。,,()(s)。[3]:,。,。2　2.1　　(1)(2),:①1080;②1550;③:、、;,,。图1　小鼠咳嗽次数的频数图图2　小鼠咳嗽潜伏期的频率图2.2　　1。、;,。,。　　1　(x±s)()()(s)()()(s)45924.59±21.3529.28±12.9721629.04±19.19＊＊28.69±10.3243924.73±18.5129.43±13.6026927.51±14.51＊29.11±9.8889824.66±20.0029.35±13.2348528.19±16.76＊＊28.92±10.07　　＊P0.05,＊＊P0.012.3　　;,,。3　3.1　,,。3.2　,,,,,,,;,,,,。3.3　,,,:①1080;②1550;③:、、;,,。1　,,..:,2001∶1362～13662　..:,2006∶428～4483　,,,..,2006,23(6)∶338～391杜远东1,赵玉珍2,赵宏伟2,郑　旭2,陈　芳2,王莉芳2(1陕西中医学院药学院,咸阳　712046;2陕西省食品药品检验所,西安　710061)　　　　:采用过敏试验法与降压物质检查法评价舒血宁注射液的安全性。:以豚鼠为受试对象进行过敏性试验研究,以猫为受试对象进行降压物质试验研究。:舒血宁注射液过敏试验的给药剂量为临床用量5.8倍、11.6倍、23.2倍,后第1512010;26(5)