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ID:36565732
大小:66.68 KB
页数:6页
时间:2019-05-12
《豚鼠哮喘模型和哮喘患者气管上皮细胞内皮素1和内皮素转化酶mRNA表达》由会员上传分享,免费在线阅读,更多相关内容在应用文档-天天文库。
1、豚鼠哮喘模型和哮喘患者气管上皮细胞内皮素-1和内皮素转化酶mRNA表达作者:何翔梁永杰汪蜀郭忠良【关键词】内皮素转换酶【摘要】目的通过逆转录-DNA聚合酶链反应(RT-PCR)技术检测豚鼠哮喘模型气管上皮内皮素-1(ET-1)mRˉNA表达,检测哮喘患者气管上皮内皮素-1(ET-1)和内皮素转换酶(ECE)mRNA表达。方法(1)动物实验部分:Hartley株豚鼠用卵蛋白腹腔注射致敏,雾化吸入激发,提取气管上皮细胞RNA,逆转录将RNA逆转为cDNA,PCR扩增DNA。(2)临床实验部分:设对照组和哮喘组。一步法提取总RNA,逆转录合成cDNA,PCR扩增β-actin片段、ET-1片段
2、和ECE片段。PCR产物的鉴定和半定量。计算ET-1/β-actin值和ECE/β-actin值。结果(1)动物模型检测:在电泳缓冲液内,紫外灯下可见ET-1为333bp左右条带,与标记DNA条带相对应。(2)临床实验:哮喘组ET-1mRNA表达量明显高于对照组(P<0.05)。哮喘组ECEmRNA表达量与对照组比较差异无显著性(P>0.05)。结论RT-PCR技术检测出卵蛋白致敏激发的豚鼠气管上皮细胞ET-1mRNA表达。哮喘患者支气管粘膜ET-1mRNA表达明显增高,而ECEmRNA表达量与对照组比较差异无显著性。关键词RT-聚合酶链反应内皮素-1内皮素转换酶6【Abst
3、ract】ObjectiveTodetectthemRNAexpressionsofendothelin-1intrachealepitheliumcellfromguineapigasthmamodelbyreversetranscriptionalpolymerasechainreaction,todetectthemRNAexpressionsofenˉdotheline-1andendothelinconvertingenzymeintrachealepitheliumcellfromasthmapatient.Methods(1)Animalexperimentdepartme
4、nt:sensitizetheguineapigbyinjectovi-proteinintotheabdominalcavity,excitationbyaspiratetheovi-proteinsolutions,extractRNAfromtrachealepitheliumcell,reversetranscribetheRNAtocDNA,amplifyDNAbypolymerasechainreaction.(2)Clinicaldepartment:layingtheasthemagroupandthecontrolgroup,extractRNAfromtracheal
5、epitheliumcell,reversetranscribetheRNAtocDNA,amplifyβ-actin、ET-1andECEfragmentbypolymerasechainreaction.evaluateandsemi-quantifythePCRproduct,calculateET-1/β-actinandECE/β-actin.Results(1)Animalexperiment:wecanseetheET-1isabouta333bpstrapintheelectrophoresissolutionundertheinfraredlamp,correspond
6、withthemarkingstrap.(2)Clinicaldepartment:ThemRNAexpressionsofenˉdothelin-1oftheasthemagroupisgreathighthanthecontrolgroup(P<0.05),thedifferenceofmRNAexpressionsofECEisnotsignificantattwogroups(P>0.05).ConclusionItcandetectthemRNAexpressionsof6endothelin-lintrachealepitheliumcellfromguineap
7、igasthmamodelbyreversetranscriptionalpolymerasechainreaction.ThemRNAexpressionsofendothelin-1oftheasthemagroupisgreathighthanthecontrolgroup,ThediffereceofmRNAexpressionsofECEisnotsignificantattwogroups.Keywordsreverse
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