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ID:33724811
大小:2.41 MB
页数:23页
时间:2019-02-28
《大鼠缺血性脑水肿早期aqp4mrna表达的动态变化分析》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、(P<0。05)。实验同时显示脑水含量与AQP4mRNA表达呈正相关(r=O。871,P2、·eartysta殳eO薹iscnemicbrainedemainratsObjectiveToinvestigatetheexpressionofaquaporin-4(AQP4)mRNAintheearlystageofcerebraledemafollowingischemiainratsandelucidatetherelationshipbetweenthedynamicchangesofbrainwatercontentandexpressionofAQP4mRNA.TheresultsmayrevealthefunctionandeffectofAQP4inearly3、stageofischemicbrainedemaandbebeneficialtofindanewapproachforthetherapyofcerebraledema.Methodsl。Materials1.1Animalandgrouping:66healthyWistarratswererandomlyassignedtopseudo—operationgroup,operationgroup,TTCstaininggroup.EachgroupWasdividedintosixsub-groupaccordingtodifferenttimepoints(1h,3h,64、h,12h,18h,24h).Everysubgroupcontainedfiverats.1.2Mainagentsandapparatus:RNAPCRKit,PCRamplificationsystem,gelimaginganalysissystem,ultravioletspectrometer.1,3Animalmodel:ArevisedmodelofLinehitchlawaccordingtoZeaLonga.1.4TTCstaining:Itdemonstratesthemodelissucceed.2。Indexdetection2.1Brainwaterco5、ntent:detenmnedbywet-dryweight.2.2DetectingofAQP4mRNAinbraintissue:11∽expressionofAQP4mRNAwasassessedbyRT—PCR.RNA1uLwasextractedfromthebraintissue,addingrelatedreagents。TheeDNAwassynthesizedbymRNAthroughreversetranscription,mixedtheeDNAwithAQP4specificprimersandrelatedreagents,during40secondsw6、ith94"C,1minutewith50℃,lminutewith72"C,after35cycles,extension7minutes晰也72℃,theamplifiedproductsofAQP4Wasmixedwithequal13-actinamplifiedproducts,thenbrought5ulmixedproductsin2%agarosegelbed,electrophoresis,the4ratio(GrayOftheratio)ofAQP4mRNAtotheopticaldensityintegralof13-actinelectrophoresisp7、roductswereobservedandcalculatedbygelimaginganalysissystemobservationandcalculation,whichdeterminedtheexpressionlevelAQP4mRNA.3。StatisticalprocessingAlldatawereanalyzedbySPSSI3.0statisticalsoft.ThedatafromeverygroupwerepresentedbyX+S,th
2、·eartysta殳eO薹iscnemicbrainedemainratsObjectiveToinvestigatetheexpressionofaquaporin-4(AQP4)mRNAintheearlystageofcerebraledemafollowingischemiainratsandelucidatetherelationshipbetweenthedynamicchangesofbrainwatercontentandexpressionofAQP4mRNA.TheresultsmayrevealthefunctionandeffectofAQP4inearly
3、stageofischemicbrainedemaandbebeneficialtofindanewapproachforthetherapyofcerebraledema.Methodsl。Materials1.1Animalandgrouping:66healthyWistarratswererandomlyassignedtopseudo—operationgroup,operationgroup,TTCstaininggroup.EachgroupWasdividedintosixsub-groupaccordingtodifferenttimepoints(1h,3h,6
4、h,12h,18h,24h).Everysubgroupcontainedfiverats.1.2Mainagentsandapparatus:RNAPCRKit,PCRamplificationsystem,gelimaginganalysissystem,ultravioletspectrometer.1,3Animalmodel:ArevisedmodelofLinehitchlawaccordingtoZeaLonga.1.4TTCstaining:Itdemonstratesthemodelissucceed.2。Indexdetection2.1Brainwaterco
5、ntent:detenmnedbywet-dryweight.2.2DetectingofAQP4mRNAinbraintissue:11∽expressionofAQP4mRNAwasassessedbyRT—PCR.RNA1uLwasextractedfromthebraintissue,addingrelatedreagents。TheeDNAwassynthesizedbymRNAthroughreversetranscription,mixedtheeDNAwithAQP4specificprimersandrelatedreagents,during40secondsw
6、ith94"C,1minutewith50℃,lminutewith72"C,after35cycles,extension7minutes晰也72℃,theamplifiedproductsofAQP4Wasmixedwithequal13-actinamplifiedproducts,thenbrought5ulmixedproductsin2%agarosegelbed,electrophoresis,the4ratio(GrayOftheratio)ofAQP4mRNAtotheopticaldensityintegralof13-actinelectrophoresisp
7、roductswereobservedandcalculatedbygelimaginganalysissystemobservationandcalculation,whichdeterminedtheexpressionlevelAQP4mRNA.3。StatisticalprocessingAlldatawereanalyzedbySPSSI3.0statisticalsoft.ThedatafromeverygroupwerepresentedbyX+S,th
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