一种病毒性哮喘模型制作方法的建立及评价

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1、一种病毒性哮喘模型制作方法的建立及评价【摘要】目的评价用呼吸道合胞病毒(RSV)诱导鸡卵白蛋白(OVA)致敏，建立小鼠急性病毒性哮喘模型的方法及效果。方法BALB/c小鼠100只，随机分为磷酸盐缓冲液(PBS/PBS)对照组、PBS/RSV组、RSV/RSV组、OVA/PBS组和OVA/RSV组5组。应用OVA多点注射致敏、OVA气道雾化吸入结合RSV滴鼻激发复制急性病毒性哮喘模型。观察小鼠哮喘急性发作症状；动物体描箱法测定乙酰胆碱(Ach)激发条件下气道反应性(以肺阻力RL表示)；支气管肺泡灌洗液(BALF)作细胞分类计数；HE染色观察

2、肺组织病理变化。结果①以45、135、405μg/mLAch激发时，与其他各组比较，OVA/RSV组RL均显著升高(P均<0.01)；②BALF中炎性细胞分类计数，与其他各组比较，OVA/RSV组的细胞总数、淋巴细胞、嗜酸性粒细胞、中性粒细胞均显著升高(P均<0.01)；③OVA/RSV组的肺组织病变程度重于其他各组(P均<0.01)。结论RSV诱导OVA致敏的方法建立小鼠急性病毒性哮喘模型是成功的。【关键词】呼吸道合胞病毒支气管哮喘急性模型方法及评价　　ABSTRACT:ObjectiveToreproduceatyp

3、eofacuteviralasthmamodelinovalbumin(OVA)sensitizedBALB/cmicebyinductionwithrespiratorysyncytialvirus(RSV).MethodsOnehundredmicewererandomlydividedintofivegroups:PBS/PBS14controlgroup,PBS/RSVgroup,RSV/RSVgroup,OVA/PBSgroupandOVA/RSVgroup.Theasthmamodelwasreproducedbysensi

4、tizationwithmultipointintraperitonealinjectionofOVA,followedbyrepeatedinhalationofOVAcombinedwithrepeatedintranasalinstillationofRSV(1.0×106.5TCID50/mLin50μL).Asthmaticsymptomswereobserved.Thechangesinairwayresponsivenesswererepresentedbylungresistance(RL)stimulatedbyacet

5、ylchdine(Ach);Cellsinbronchoalveolarlavagefluid(BALF)werecountedandlunghistopathologywasevaluatedbyHEstaining.Results①ThedifferencesinRLinOVA/RSVgroupweresignificantwhencomparedwiththoseinPBS/PBScontrolgroup,PBS/RSVgroup,andRSV/RSVgroup,OVA/PBSgroupwhenstimulatedwithAch(4

6、5,135,405μg/mL)(P<0.01,respectively).AsthmasymptomsweremoresevereinOVA/RSVgroupcomparedwiththoseinOVAgroup.②ThedifferentialcountofsexualcellsinBALF,thedifferencesinthenumberoftotalcellularscore,lymphocytes,eosinophils,andneutrophilsinOVA/RSVgroupweresignificantwhencomp

7、aredwiththoseintheothergroups(P<0.01,respectively).③HEstaining:ThedifferencesinthechangesoflunghistopathologyinOVA/RSVgroupweresignificantwhencomparedwiththoseintheothergroups(P<0.01,respectively).ConslusionAnanimalmodelofacute14viralasthmamodelissuccessfullyreprodu

8、cedbynasalinoculationwithRSVinOVAsensitizedBALB/cmice.　　KEYWORDS:respiratorysyncytialvirus;asth