肾凝血酶原：在大鼠模型中使用逆转录-聚合酶链反应对mRNA进行定量分析。

Renal prothrombin: quantification of mRNA using reverse transcription-polymerase chain reaction in a rat model.

作者信息

Grover Phulwinder K, Miyazawa Katsuhito, Ryall Rosemary Lyons

机构信息

Urology Unit, Department of Surgery, Flinders Medical Centre, Bedford Park, South Australia 5042, Australia.

出版信息

Electrophoresis. 2004 Mar;25(6):797-803. doi: 10.1002/elps.200305731.

DOI:10.1002/elps.200305731

PMID:15004838

Abstract

Attempts to quantify renal prothrombin (PT) have failed due to interference of its hepatic counterpart. In order to gauge PT synthesized by the kidney, the expression of PT mRNA was compared in renal and hepatic tissues of rats. PT mRNAs were quantified, using quantitative competitive reverse transcription-polymerase chain reaction (RT-PCR), from RNA extracts of kidneys and livers of ten hooded Wistar rats. To control variations in sample preparation, the amount of PT transcript in each sample was normalized to the amount of its beta-actin transcript. The median ratio of PT to beta-actin transcript of 0.0011 (with a mean +/- standard deviation (SD) of 0.0010 +/- 0.0002, range 0.0007-0.0014) in the renal tissues was significantly (p < or = 0.001) lower than its corresponding value of 0.3669 (with a mean +/- SD of 0.3729 +/- 0.0716, range 0.2718-0.4675) in the hepatic tissues. Thus, expression of PT mRNA in the rat kidney was almost 300-fold less as compared with its expression in hepatic tissue.

摘要

由于肝脏中凝血酶原（PT）的干扰，对肾脏中PT进行定量的尝试失败了。为了评估肾脏合成的PT，比较了大鼠肾脏和肝脏组织中PT mRNA的表达。使用定量竞争性逆转录 - 聚合酶链反应（RT-PCR）对十只带帽Wistar大鼠的肾脏和肝脏RNA提取物中的PT mRNA进行定量。为了控制样品制备中的差异，将每个样品中PT转录本的量相对于其β-肌动蛋白转录本的量进行标准化。肾脏组织中PT与β-肌动蛋白转录本的中位数比值为0.0011（平均值±标准差（SD）为0.0010±0.0002，范围为0.0007 - 0.0014），显著低于（p≤0.001）其在肝脏组织中的相应值0.3669（平均值±SD为0.3729±0.0716，范围为0.2718 - 0.4675）。因此，大鼠肾脏中PT mRNA的表达与其在肝脏组织中的表达相比几乎低300倍。