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

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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