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时间:2018-11-20
《豚鼠哮喘模型和哮喘患者气管上皮细胞内皮素》由会员上传分享,免费在线阅读,更多相关内容在学术论文-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-act
2、in片段、ET-1片段和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
3、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【Abstract】ObjectiveTodetectthemRNAexpressionsofendothelin-1intrachealepitheliumcellfromguineapigasthmamodelbyreversetranscriptionalpolymerasechainreaction,todetectthemRNAexpressionsofenˉdotheline-1andendothelinconvertingenzymeintrachealepit
4、heliumcellfromasthmapatient.Methods(1)Animalexperimentdepartment:sensitizetheguineapigbyinjectovi-protEinintotheabdominalcavity,excitationbyaspiratetheovi-protEInsolutions,extractRNAfromtrachealepitheliumcell,reversetranscribetheRNAtocDNA,amplifyDNAbypolymerasechainreact
5、ion.(2)Clinicaldepartment:layingtheasthemagroupandthecontrolgroup,extractRNAfromtrachealepitheliumcell,reversetranscribetheRNAtocDNA,amplifyβ-actin、ET-1andECEfragmentbypolymerasechainreaction.evaluateandsemi-quantifythePCRproduct,calculateET-1/β-actinandECE/β-actin.Resul
6、ts(1)Animalexperiment:p,correspondarkingstrap.(2)Clinicaldepartment:ThemRNAexpressionsofenˉdothelin-1oftheasthemagroupisgreathighthanthecontrolgroup(P<0.05),thedifferenceofmRNAexpressionsofECEisnotsignificantattRNAexpressionsofendothelin-lintrachealepitheliumcellfromg
7、uineapigasthmamodelbyreversetranscriptionalpolymerasechainreaction.ThemRNAexpressionsofendothelin-1oftheasthemagroupisgreathighthanthecontrolgroup,ThediffereceofmRNAexpressionsofECEisnotsignificantatterasereactionendothelin-1endothelinconvertingenzyme内皮素-1(ET-1)是至今所发现的最强
8、烈的支气管平滑肌收缩物质之一,并有促进成纤维细胞和平滑肌细胞增殖,以及促进粘膜下腺体分泌的作用[1]。研究发现ET-1参与哮喘的发病和发展[2,3],哮喘患者和动物模型的血浆[4]或气管上皮ET-1水平明显
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