豚鼠哮喘模型和哮喘患者气管上皮细胞内皮素.docVIP

　　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-actin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（Plt;0.05）。哮喘组ECEmRNA表达量与对照组比较差异无显著性（Pgt;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 【Abstract】 Objective To detect the mRNA expressions of endothelin-1in tracheal epithelium cell from guinea pig asthma model by reverse transcriptional polymerase chain reaction，to detect the mRNA expressions of enˉdotheline-1and endothelin converting enzyme in tracheal epithelium cell from asthma patient.Methods （1）Animal experiment department:sensitize the guinea pig by inject ovi-protEin into the abdominal cavity，excitation by aspirate the ovi-protEIn solutions，extract RNA from tracheal epithelium cell，reverse transcribe the RNA to cDNA，amplify DNA by polymerase chain reaction.（2）Clinical department:laying the asthema group and the control group，extract RNA from tracheal epithelium cell，reverse transcribe the RNA to cDNA，amplifyβ-actin、ET-1and ECE fragment by polymerase chain reaction.evaluate and semi-quantify the PCR product，calculate ET-1/β-actin and ECE/β-actin.Results （1）Animal experiment:p，correspond arking strap.（2）Clinical department: The mRNA expressions of enˉdothelin-1of the asthema group is great high than the control group（Plt;0.05），the difference of mRNA expressions of ECE is not significant at tRNA expressions of endothelin-l in tracheal epithelium cell from guinea pig asthma model by reverse transcriptional polymerasechain reaction.The mRNA expressions of endothelin-1of the asthema group is great high than the control group，The differece of mRNA expressions of ECE isn