半定量RT－PCR方法在基因表达研究中的应用

SEMI-QUANTITATIVE RT-PCR METHOD AND ITS APPLICATION IN THE STUDY ON GENE EXPRESSION

目的：定量检测Ｂ型内皮素受体（ＥＴＢ－Ｒ）基因在大鼠心、肝、肺、肾等器官及培养的肾小球系膜细胞（ＭＣ）的表达。方法：半定量逆转录。多聚酶链反应技术（ＳｑＲＴ－ＰＣＲ），并以ＮｏｒｔｈｅｒｎＥＰ印迹法作对照。结果：ＥＴＢ－Ｒ基因在上述器官及ＭＣ均有表达，而表达水平依次为肺、心、肾和肝。结论：ＥＴＢ－Ｒ在不同器官有不同水平的基因表达，说明内皮素对不同器官的效应不同；ＭＣ是内皮素在肾内发挥作用的靶细胞。

Objective:Quantitative analysis of expression of endothelin B receptor(ETB-R)gene in rat heart,liver,lung,kidney and cuItured glomerular mesangial cells(MC)were observed. Methods:Se-mi-quantitative reverse transcription and polymerase chain reaction(SqRT-pCR)was used and com-pared with Northern blot hybridization. Results:EIB-R gene was expressed in all the above organs and MC,and highest expression level in the lung,moderate in the heart and kidney, but lowest in the liver. Conclusion:Different expression level of ETB-R gene in various organs imply that the effects of en-dothelin on different organs were not similar.The MC were target cells of endothelin in kidney.